Three-spined stickleback: an emerging model in environmental endocrine disruption.

The three-spined stickleback, a small teleost species with habitats that range from full marine to fresh water bodies across the whole Northern hemisphere, has a number of advantages for endocrine disruption research. It is the only teleost species with an unambiguous biomarker for androgens, the presence of the glue protein spiggin in the male kidney, which can be measured by enzyme-linked immunosorbent assay (ELISA). The androgen assay has been adopted to detect antiandrogens in two different ways and an homologous ELISA for stickleback vitellogenin is also available. DNA markers for molecular sex determination are available; thus, sex ratios can also be used for in situ biomonitoring. In addition, the critical period of sexual differentiation has been determined and the occurrence of intersex fish has been reported several times. The species full genome sequence is almost complete. All aspects of stickleback biology (ecology, evolution, behavior, physiology, endocrinology) are well documented. In European waters, the stickleback is the only fish that can bring laboratory and field studies together and allow the true impact of endocrine disruptors on fish populations to be evaluated.     

The artistic three-spined stickleback (<Emphasis Type="Italic">Gasterosteous aculeatus</Emphasis>)

We investigated whether female three-spined sticklebacks ( Gasterosteus aculeatus) are more attracted to a male whose nest contains colourful artificial material than to a nest that only contains algae. The purpose was to examine whether nest characters are of importance for female choice in the three-spined stickleback. Most studies on female preference in this species have focused on male morphological traits. In our study, on a marine population of three-spined stickleback on the West coast of Sweden, we found that males marked their nests' entrance with a deviant colour of algae — an apparent decoration. Moreover, males also decorated their nests with shiny and colourful foil sticks and spangles when provided with such, thereby apparently reducing the nest camouflage. In a mate choice experiment, females were more attracted to males with nests containing sticks and spangles than to males with undecorated nests. Nest decoration may be important for the evolution of paternal care in this species: if males advertise their paternal skills by ornamenting their nest they will receive more matings than males with dull nests.     

Contrasting population structures in two sympatric fishes in the Baltic Sea basin

Detailed multispecies studies on the patterns of genetic variability and differentiation in marine environments are still rare. Using mitochondrial and nuclear genetic markers, we compared genetic variability and population structuring of threespine (Gasterosteus aculeatus) and ninespine (Pungitius pungitius) sticklebacks from the same eleven marine and six freshwater locations within the Baltic Sea basin. Analyses of both marker types revealed a significantly lower degree of genetic structuring in both marine and freshwater populations of threespine than those ninespine sticklebacks. Isolation-by-distance (IBD) was detected across the marine populations in both species, suggesting spatially limited gene flow. However, the levels of genetic diversity and differentiation across the localities were uncorrelated between the two species in both marine and freshwater environments. Accordingly, estimates of effective population sizes were larger and migration rates were higher for three- than for ninespine sticklebacks. Hence, ninespine stickleback populations from the Baltic Sea basin appear to be subject to stronger genetic drift than sympatric threespine sticklebacks, and the proximate reason for this difference is likely to be found from autecological differences between the two species. In accordance with the earlier studies, genetic variability was higher and the degree of genetic differentiation was lower in marine than in freshwater populations in both species.     

Geographic variation in Loligo forbesi in the Northeast Atlantic Ocean: analysis of morphometric data and tests of causal hypotheses

Geographic variation in the squid Loligo forbesi was investigated using multivariate analysis of morphometric and meristic characters in samples of squid taken from 13 localities in the Northeast Atlantic Ocean. Two character sets, body morphometrics and beak morphometrics, indicated similar patterns of variation, with squid from the Azores differing markedly from those on the continental shelf. No consistent pattern was apparent in meristic data. Partial Mantel tests indicated that similarity matrices for morphological data were significantly correlated with distance matrices for (a) geographic proximity, (b) whether the capture site was on the continental shelf or the Azorean bank, and (c) (beak data only) average seasurface temperature at site of capture. Partial Mantel tests on allozyme data for the same individuals support hypothesis (b). The results suggest that L. forbesi in the Azores may reasonably be regarded as a distinct stock, differing significantly from L. forbesi on the continental shelf.     

Development of a new antifouling method for a marine cooling water system

Variations in meristic characters occur within and between samples of milkfish Chanos chanos (Forsskal) collected on June 1977 from five localities in Philippine waters. The unbranched anal and ventral fin rays are the most plastic, and vertebral number is the most stable of nine meristic characters examined. The number of ventral fin rays is the only meristic feature which differed consistently enough to suggest distinct population groups: South China Sea, comprising samples from Ilocos and Panay Island; Pacific Ocean from Bicol; and Celebes Sea, from Zamboanga and Davao. A discriminant analysis which incorporated generalized distance (Mahanalobis D²) and percent overlap of a reduced set of characters indicated several morphometric subgroups of milkfish in Philippine waters, although geographic subgroups were not clearly demonstrated.SEAFDEC Contribution No. 218     

Diversionary displays of paternal stickleback

Summary In Crystal Lake, British Columbia, male threespine stickleback (Gasterosteus aculeatus) perform a diversionary display that deters cannibalistic conspecifics from entering their territories and destroying their nests or consuming offspring. In this population, the display consists of males swimming directly out of their territories and rooting aggressively in the substratum, mimicking stickleback feeding on nests of others, or digging for nesting materials. The display is elicited solely by conspecifics, and usually by benthic foraging groups of 2–300 individuals. It usually is elicited when groups are approaching the territory directly. Otherwise the male typically joins the group or remains still in his territory until the group leaves. This observation suggests that males are capable of discriminating conditions under which the display is most likely to be successful. The display is absent in a population in Garden Bay Lake, British Columbia, in which benthic foraging groups are absent. In Crystal Lake the males themselves do not risk predation by the groups, and the tendency of males to perform the display does not change with reproductive condition. Comparison with diversionary displays performed in two additional allopatric populations suggests that diversionary displays in this species incorporate elements of other behavioral sequences displaced to the display, and that variation in the structure of the display exists among populations.     

Sympatric species of threespine stickleback differ in their performance in a spatial learning task

Increasing evidence suggests that cognitive function is under selection in diverse taxa and that this results in different cognitive abilities in taxa experiencing different selective environments. For example, species inhabiting spatially complex environments might be expected to have good spatial learning ability. We investigated whether local habitat conditions influence learning by comparing the performance of two recently diverged species of threespine stickleback (Gasterosteus aculeatus complex) in a spatial learning task. The two species reside sympatrically in several lakes. Benthics occupy the spatially structured vegetated littoral zone, whereas limnetics occupy the spatially homogenous open-water pelagic zone. We trained fish to locate a hidden reward in a T-maze and asked whether they did so by learning a body-centred pattern of movement (turn left or right) or by using plant landmarks. Both benthics and limnetics used turn and landmark cues, but benthics learnt the task almost twice as quickly as limnetics. This difference is consistent with the hypothesis that benthic and limnetic sticklebacks are equipped with spatial learning abilities well suited to the spatial complexity of their littoral and pelagic habitats. Our findings add to the understanding of the evolution of learning.     

<Emphasis Type="Italic">Patella aspera</Emphasis> and<Emphasis Type="Italic"> P. ulyssiponensis</Emphasis>: genetic evidence of speciation in the North-east Atlantic

The group of subspecies of Patella ulyssiponensis, described by Christiaens, was widely known as Patella aspera until recently. The group extends throughout the Mediterranean, on all Macaronesian islands, along the North African coast, and in Europe, as far north as southern Norway. Throughout its range it displays great variation in shell sculpture and colour. The aim of this work was to re-examine the various subspecies proposed by Christiaens and, genetically, to test the hypothesis that European continental populations belong to a different species than the one composed by Macaronesian (north-west African) populations. In the present work, this group was studied by allozyme electrophoresis of 21 loci and by six morphological variables. The monomorphic locus of malate dehydrogenase (Mdh-1) was found to be diagnostic for distinguishing European continental populations from those of north-west African archipelagos, confirming the specific status of both groups. The allele observed at this locus in the Macaronesian populations was novel, while European continental populations showed a plesiomorphic one, shared with all other north-west Atlantic patellids with the exception of Patina pellucida, suggesting a more recent origin of the Macaronesian species from the continental forms. Both species showed a genetic identity of 0.730±0.061, which allowed rough estimations of 6.5–3 Mya since speciation. The subspecies Patella ulyssiponensis deserta described by Christiaens was not confirmed by our genetic data. Other loci (glyceraldehyde-3-phosphate dehydrogenase and the second locus of lactate dehydrogenase) were partially diagnostic, in which both species showed different most common alleles. Morphologically, both species are easily recognised by shell characters and the results agree with previous findings, that continental populations are more homogeneous in shell morphology and radula characters than populations from the Macaronesia. This work supports retention of the earliest valid name, Patella ulyssiponensis Gmelin, with Lisbon, Portugal as type locality, for the European continental species, and Patella aspera Röding for the Macaronesian populations. Population subdivision within species was measured by theta, the estimator of F_st, showing in both P. aspera and P. ulyssiponensis a high degree of genetic structuring (=0.226 and 0.182, respectively) mostly explained by the large distances separating the populations within species.Communicated by J.P.Thorpe, Port Erin     

Nodularin concentrations in Baltic Sea zooplankton and fish during a cyanobacterial bloom

Toxic cyanobacterial blooms, dominated by Nodularia spumigena, are a recurrent phenomenon in the Baltic Sea during late summer. Nodularin, a potent hepatotoxin, has been previously observed to accumulate on different trophic levels, in zooplankton, mysid shrimps, fish as well as benthic organisms, even in waterfowl. While the largest concentrations of nodularin have been measured from the benthic organisms and the food web originating from them, the concentrations in the pelagic organisms are not negligible. The observations on concentrations in zooplankton and planktivorous fish are sporadic, however. A field study in the Gulf of Finland, northern Baltic Sea, was conducted during cyanobacterial bloom season where zooplankton (copepod Eurytemora affinis, cladoceran Pleopsis polyphemoides) and fish (herring, sprat, three-spined stickleback) samples for toxin analyses were collected from the same sampling areas, concurrently with phytoplankton community samples. N. spumigena was most abundant in the eastern Gulf of Finland. In this same sampling area, cladoceran P. polyphemoides contained more nodularin than in the other areas, suggesting that this species has a low capacity to avoid cyanobacterial exposure when the abundance of cyanobacterial filaments is high. In copepod E. affinis nodularin concentrations were high in all of the sampling areas, irrespective of the N. spumigena cell numbers. Furthermore, nodularin concentrations in herring samples were highest in the eastern Gulf of Finland. Three-spined stickleback contained the highest concentrations of nodularin of all the three fish species included in this study, probably because it prefers upper water layers where also the risk of nodularin accumulation in zooplankton is the highest. No linear relationship was found between N. spumigena abundance and nodularin concentration in zooplankton and fish, but in the eastern area where the most dense surface-floating bloom was observed, the nodularin concentrations in zooplankton were high. The maximum concentrations in zooplankton and fish samples in this study were higher than measured before, suggesting that the temporal variation of nodularin concentrations in pelagic communities can be large, and vary from negligible to potentially harmful.     

Generalist cuckoo bees (Hymenoptera: Apoidea: Sphecodes) are species-specialist at the individual level

Intensive and incessant arms races between a parasite and its host are generally expected to lead to parasite specialization. Nevertheless, some parasitic species still successfully attack wide spectra of hosts. One of the solutions to the evolutionary enigma of the long-term existence of generalist parasites is their specialization at an individual level, a phenomenon well known, e.g., in European common cuckoo. Over its range, it parasitizes a number of bird species; however, individual females are mostly specialists possessing adaptations to a particular host species. In this study, we test the possibility of individual specialization in generalist cuckoo bees, the insect counterparts of avian cuckoos. Females of cuckoo bees lay each egg into a single brood cell in the nests of other bee species. The host’s offspring is destroyed by the parasitic female or later by her larvae, which feed on pollen supplies accumulated by the host. Both studied cleptoparasitic bees (Sphecodes ephippius and Sphecodes monilicornis) are widely distributed in Europe, where they have been reported to use broad host spectra. We recorded several host species (including some previously unknown) for both cuckoo bee species, and confirmed that these parasites are indeed generalist even at a small local scale. However, we demonstrate that exactly as in the avian cuckoos, each female in both species of generalist bee parasites tends to attack just one host species.     

Phylogeny and biogeographic history of hake (genus Merluccius), inferred from mitochondrial DNA control-region sequences

 Phylogenetic analyses of the left domain of the mitochondrial DNA control-region sequence have been used to examine the relationships among species of the genus Merluccius (Rafinesque, 1810), and to compare these with hypotheses based on morphological, meristic and allozyme characters. Analysis of aligned sequences revealed that transition bias was much lower than in mammalian mtDNA, and that nucleotide composition of control-region sequences was biased toward A and T. We have roughly calibrated a molecular clock for the genus, based on the rise of the Isthmus of Panamá, which is believed to have created a barrier to dispersal between marine species of the Atlantic and Pacific Oceans. Our mtDNA-based phylogeny was highly congruent with allozyme-based phylogenies, but poorly so with a previously described phylogeny based on morphology. Specifically, our phylogeny resolved two well-supported principal clades, one of American (west Atlantic and east Pacific) species and the other of Euro–African (east Atlantic) species. This suggests an evolutionary history during which the ancestral lineage of Merluccius was divided between two geographic regions, with subsequent dispersal and vicariant events resulting in the evolution and distribution of extant taxa. However, the relationships between some taxa within the American clade could not be resolved. We suggest that this is consistent with an hypothesis of a rapid origin and radiation of these taxa. Received: 12 December 1998 / Accepted: 15 October 1999     

Population structure of the widely dispersing marine bryozoan Membranipora membranacea (Cheilostomata): implications for population history, biogeography, and taxonomy

Morphologically plastic, cryptic, or geographically widespread species pose similar challenges to the evolutionary biologist: their taxonomic status is often unclear yet must be known to study almost any aspect of their biology, ecology, evolution, or biogeography. The marine bryozoan Membranipora membranacea (L.) is morphologically plastic and geographically widespread in temperate oceans of the Northern and Southern Hemispheres, and its taxonomy is unclear. This study examined genetic relationships among allopatric populations and sympatric morphs of this species, or species complex. Colonies were collected from 1992 to 1995. Allozymes were used to elucidate the relationships among four widely separated populations, two in the North Atlantic and two in the North Pacific Ocean. Allozymes and mtDNA sequencing were used to clarify the genetic relationships among three sympatric morphs that might correspond to the species M. villosa Hincks and M. membranacea in the northeastern Pacific (Washington State). Populations in the North Atlantic and North Pacific had no fixed allelic differences at the loci tested but were separated by an average Nei's genetic distance of 0.581, suggesting their near-sibling species status. Populations from Friday Harbor (Washington) and Catalina Island (California) were not significantly differentiated, which was attributed to high gene flow. Populations on either side of the North Atlantic were genetically indistinguishable, which is most likely due to the recent establishment of the West Atlantic populations from European founders. At Friday Harbor, sympatric morphs varying in their spination and spine inducibility were genetically indistinguishable, supporting the hypothesis that M. villosa is an induced phenotype of M. membranacea and not a distinct species in the northeastern Pacific. Since such phenotypic plasticity is common in cheilostome bryozoans, the morphospecies concept must be used with caution. Received: 31 August 1998 / Accepted: 10 August 1999     

Geographic and reproductive isolation in the marine Harpacticoid Copepod Tisbe

Within the genus Tisbe (Copepoda: Harpacticoida), there is a wide range of reproductive isolation. In the European area, in addition to species where (between geographic populations) the reproductive barrier is practically absent, there are others which show a varying degree of incompatibility. The present paper deals with the results of cross-breeding experiments carried out between American and European populations. The respective intrapopulation crosses served as controls. Transatlantic crosses have been attempted with the following species: T. lagunaris, T. bulbisetosa, T. holothuriae, T. battagliai, and T. clodiensis. The American and European populations of the first 4 species are interfertile, and the F₁ hybrids produce viable offspring. Moreover, the interpopulation crosses, compared to the controls, exhibit a slight superiority whose nature is, however, still unclear. The fifth species, T. clodiensis, behaves differently. The absence of viable offspring from interpopulation crosses, or the death of the hybrids before reaching the adult stage, indicate that, in this species, the barrier to gene flow is complete. Populations of T. clodiensis show, in addition, signs of relative intraspecific incompatibility. The problem of the different degree of reproductive isolation in the genus Tisbe, with special regard to the role played by ecological factors in determining the isolation patterns, is discussed.     

Inventory of benthic macroinvertebrates and zooplankton in several European Bonamia ostreae-endemic areas and their possible role in the life cycle of this parasite

Bonamia ostreae is a protozoan parasite of the native oyster, Ostrea edulis. The parasite can be transmitted directly from infected to uninfected oysters. Though direct transmission of the parasite can occur between oysters it is unclear if this represents the complete life cycle of the parasite, and the involvement of a carrier/reservoir cannot be ruled out. A survey of benthic macroinvertebrate and zooplankton survey was carried out in the North Channel, Cork Harbour, Ireland, an area where B. ostreae is endemic and the parasite has been present since the 1980s. This was compared to several European surveys of benthic macroinvertebrate and zooplankton species found in other B. ostreae endemic areas. The objective was to identify any species that were common to these sites and could act as possible carrier/reservoir hosts for the pathogen. Species that were found to be common in two or more sites were included in two comparison tables. These species will be targeted in future studies investigating the possible role of macroinvertebrates and zooplankton in the complete life cycle of B. ostreae.     

Adult sex ratio affects divorce rate in the monogamous endoparasite Schistosoma mansoni

“Divorce” (mate switching) rate is known to vary largely both between and within socially monogamous species. Although the adult sex ratio can have an important influence on mating patterns, very few studies have investigated the influence of sex ratio on divorce rate in monogamous species, and even less so from an experimental point of view. In addition, most studies on the causes and consequences of divorce have been performed on vertebrate species, whereas data for invertebrate monogamous species remain scarce. Schistosoma mansoni is a monogamous endoparasite with a complex life cycle characterized by asexual reproduction in the intermediate host and sexual reproduction in the definitive host. In the wild, populations of S. mansoni inside their definitive hosts are characterized by a male-biased sex ratio. We studied the influence of experimentally varying the adult sex ratio on divorce rate in S mansoni, using controlled infections of hosts with clonal populations. The more male-biased the sex ratio was, the more the divorce rate increased, whereas no such effect was observed under a female-biased sex ratio. In this study and for the first time, we showed, by handling the sex ratio, that the divorce rate increases in adult male-biased sex ratio conditions in a monogamous species.     

Functional variation of sperm morphology in sticklebacks

Some theoretical models of sperm competition make the assumption that in fish species with external fertilisation, sperm length relates positively to swimming speed at the expense of sperm longevity. Few studies have tested this assumption. We used the three-spined stickleback, Gasterosteus aculeatus L., to study functional sperm morphology. In this study, the relationship between males’ mean sperm length and fertilisation rate was investigated in vitro in a non-competitive situation. Fertilisation at different time points after sperm release was taken into account, and sperm morphology was quantified from scanning electron microscopy images. The time series of artificial fertilisations demonstrated that males which produced sperm with a longer tail fertilised faster, but their sperm had a shorter lifespan (or activity period). It was further suggested that males that produced sperm with a larger midpiece had greater fertilisation chances later on in the fertilisation process. Thus, in sticklebacks, there exists functional variation in sperm morphology, and sperm tail length is traded off against sperm longevity (or activity).     

Niche separation in two species of hare. Metabolic costs of plant phenolics

Summary The distributions of mountain hare (Lepus timidus) and European hare (L. europaeus) overlap in central Sweden, but they occupy separate food niches in winter. In Scandinavia, the European hare is mainly a grazer while mountain hare is a predominant browser. Browse contain high amounts of secondary metabolites, such as phenols, compared to grass. This raises the question if the two hare species differ in their metabolic tolerance of plant phenols and that these differences influence their food choice.Phenolic excretion in urine increased significantly with phenolic intake in both species. Excretion of glucuronic acid conjugates, one of the major pathways of elimination of phenols in both hare species, is positively correlated to phenolic intake and excretion. However, the extent of excretion of phenolics by this route was different in the two species of hare. European hares excreted substantially more glucuronic acid per amount of phenolics than mountain hare. The phenols were metabolized to a larger extent in the mountain hare, indicating a higher detoxification capacity. From these results it is likely that European hare have a higher cost for the detoxification of plant phenols compared to mountain hare. This cost and negative effect on sodium balance when feeding on browse may prevent exploitation of forest habitats by European hares, whereas mountain hare are better able to do so.     

Using fatty acid analysis to elucidate the feeding habits of Southern Ocean mesopelagic fish

Fatty acid biomarkers were used to investigate the feeding ecology of 17 mesopelagic fish species occurring in the Southern Ocean. Fatty acid signatures of species where little or no dietary information exists were compared to fatty acid signatures of species of known diets in order to elucidate their trophic position. Principal component analysis grouped species of known diets into two clusters with amphipods and copepods comprising the main prey species, respectively. Although the majority of species of unknown diet were grouped with either of these feeding guilds, a third cluster comprising only Gymnoscopelus bolini was identified suggesting a significantly different diet for this species. Electrona antarctica also exhibited significant changes in fatty acid signatures with size. Furthermore, discriminant analysis of the four most abundant species classified species with a 90% success rate thus validating the usefulness of fatty acid signatures when trying to resolve the trophic position of species where no or little dietary information exists.     

Behavioral type–environment correlations in the field: a study of three-spined stickleback

Behavioral type–environment correlations occur when specific behavioral types of individuals are more common in certain environments. Behavioral type–environment correlations can be generated by several different mechanisms that are probably very common such as niche construction and phenotypic plasticity. Moreover, behavioral type–environment correlations have important ecological and evolutionary implications. However, few studies have examined behavioral type–environment correlations in natural populations. In this study, we asked whether some behavioral types of three-spined stickleback were more likely to occur in certain social environments (alone or in a shoal with other stickleback) or in certain microhabitats in a river (in the open or under cover). We found that individuals that were in shoals with other stickleback at the time of collection from the field emerged from a refuge more quickly compared to individuals that were found alone. In addition, fish that were alone in an open microhabitat explored more of a pool compared to fish that were alone in cover, but this difference did not occur among fish that were in shoals at the time of collection. Subsequent analyses of gut contents suggested that differences in microhabitat use were consistent over time. Our study provides some of the first evidence for behavioral type–environment correlations in a natural population of non-human animals.