Genetic analysis of potential postglacial watershed crossings in Central Europe by the bullhead (Cottus gobio L.)

Natural colonizations across watersheds have been frequently proposed to explain the present distributions of many freshwater fish species. However, detailed studies of such potential watershed crossings are still missing. Here, we investigated potential postglacial watershed crossings of the widely distributed European bullhead (Cottus gobio L.) in two different areas along the Rhine–Rhône watershed using detailed genetic analysis. The main advantage of studying bullheads vs. other freshwater fish species is that their distribution has been lightly influenced by human activities and as such, interpretations of colonization history are not confounded by artificial transplantations. The genetic analyses of eight microsatellite loci revealed strong genetic similarities between populations of both sides of the Rhine–Rhône watershed in the Lake Geneva area, giving strong evidence for a natural watershed crossing of bullheads from the upper Rhine drainage into the Rhône drainage in the Lake Geneva area likely facilitated by the retreat of the glaciers after the last glacial maximum some 20 000 years ago. Populations from the Lake Geneva basin were genetically more similar to populations from across the watershed in the upper Rhine drainage than to populations further downstream in the lower Rhône. In contrast, populations from Belfort, an area, which was not covered by ice during the last glacial maximum, showed strong genetic differentiation between populations of the upper Rhine and Rhône drainages. Based on our results on the bullhead, we propose that glacial retreat may have eased the dispersal of numerous European freshwater fish species across several geological boundaries.     

Microhabitat suitability as a tool to improve the success rate of a translocation or reintroduction,case‐study of the bullhead (Cottus perifretum) in Flanders,Belgium

Microhabitat suitability models are useful tools to enhance the reintroduction success of fish. Since 2008, a translocation and reintroduction program has been carried out in Flanders to prevent substantial loss of genetic variability in the Cottus perifretum (bullhead) population, and to meet the goals set by the Habitat Directive. To this end, habitat suitability of potential headstreams was assessed on a macrohabitat and microhabitat scale prior to the reintroduction. On a macrohabitat scale, water quality, habitat structure, food availability, and fish community were screened. Based on microhabitat models for bullhead in the summer period, microhabitat suitability was assessed in headstreams where macrohabitat characteristics showed a high potential for success. Both macro‐ and microhabitat assessment showed that reintroduction of bullheads in the Nellebeek, Bruelbeek, and Mollendaalbeek would most likely lead to self‐sustaining populations. For the Sint‐Annabeek, the microhabitat suitability model, considering depth and stream velocity, estimates that 7.5–9.5% of the headstream is suitable for bullhead. However, when substratum is taken into account, the microhabitat suitability index shows that only 4.3–5.8% of the brook is suitable. The current habitat quality and quantity in the Sint‐Annabeek is estimated to support a mean total population of 812 bullheads. However, the microhabitat suitability can be substantially improved by the artificial deposition of medium‐sized gravel (5–50 mm) and large gravel (50–100 mm). By doing so, the mean estimated population size could increase up to 1,330 individuals. Our results show that habitat improvement is necessary prior to the reintroduction of bullheads in the Sint‐Annabeek.     

Spawning habitat enhancement in the European bullhead (Cottus gobio), an endangered freshwater fish in degraded lowland rivers

Due to river regulation, the natural habitat of the European bullhead (Cottus gobio) has been degraded and often there is an apparent lack of suitable spawning substrates (hard objects like stones). Addition of artificial structures to degraded habitats may be a very promising tool for habitat enhancement. In this study, we evaluated the use of ceramic tiles as artificial spawning substrates in canalised and (remaining) meandering parts of anthropogenically perturbated lowland rivers in Flanders (northern part of Belgium). Furthermore, we examined whether water depth and velocity were important determinants for the choice of tiles in these different river trajectories. Tiles were successfully used by the bullhead as spawning substrates. In the meandering parts of the river, the number of egg deposits was significantly positively correlated with water depth, while in canalised river parts, water depth and velocity were of no importance for tile usage. In general, supplementation of rivers with artificial substrates like tiles may be a promising, inexpensive and easy-to-use enhancement technique for degraded bullhead spawning habitat.     

Postglacial colonization of Northern Europe by distinct phylogeographic lineages of the bullhead, Cottus gobio

Three major phylogeographic lineages of the cottid fish Cottus gobio (bullhead) were identified in northern Europe from mitochondrial DNA sequences and allozyme data. The largely separate freshwater distributions of the lineages demonstrate distinct postglacial colonization histories. West of the Baltic Sea, Swedish lakes were invaded from the southwest (Germany). Another, eastern lineage has colonized the inland waters northeast and east of the Baltic, from refugia in northwest Russia; this lineage comprises a distinct subgroup found only from Estonia. The third lineage, found south and southeast of the Baltic, probably descended from rivers draining to the Black Sea from the north (e.g. Dnepr). In coastal waters of the Baltic Sea, and in near-coast inland waters, the lineages are now found intermixed in various combinations. The alternating fresh- and saltwater phases of the Baltic basin have variously enabled and disabled the use of coastal waters as colonization routes. Hypotheses on the chronology of dispersal and lineage mixing can be based on the distribution of the marker genes and the paleohydrographical record. The diversity of the Fennoscandian bullhead thus comprises anciently diverged (probably mid-Pleistocene) refugial lineages that in their freshwater range constitute distinct evolutionarily significant units. The thorough mixing of the various genomic origins in and around the Baltic, however, refutes the controversial view of distinct species status for the western and eastern ('Cottus koshewnikowi') bullheads. The postglacial contact of the lineages has created new diversity that cannot be interpreted in a conventional hierarchical framework of taxonomic or conservation units.     

Introduced bullheads Cottus gobio and infection with plerocercoids of Schistocephalus cotti in the Utsjoki, an Arctic river in Finland

Bullheads Cottus gobio, first found in the River Utsjoki, an Arctic river in Finland, in 1979, were sampled at six sites in August 1991 (two of these sites were also sampled previously in June and July 1991). Mean total length (L_T) of C. gobio was 45 mm (n= 1080 fish). Overall, there were slightly more females than males, with males larger than females (mean L_T males 52·1 and females 47·6 mm). Prevalence of infection with plerocercoids of Schistocephalus cotti varied from 20·0 to 43·0% over the six sites in August. Juvenile C. gobio had a prevalence of infection of 3·5% whereas in mature fish prevalence was consistently higher in females than in males (38·8 v. 20·1%). There was one plerocercoid per infected juvenile C. gobio. In mature fish the mean number varied between 1·3 and 1·8: 49% had one worm per fish, 38% two or three worms and 13% had four to eight plerocercoids. In juvenile C. gobio, S. cotti were randomly distributed (overdispersion index 1·10), whereas occurrence was slightly aggregated in the L_T groups 40–59 and ≥60 mm (overdispersion indices both 1·90). Mean mass of infected fish was higher in all L_T groups compared to uninfected C. gobio. After removal of plerocercoids, mean mass of fish tissue remaining was lower compared with uninfected fish for the two L_T groups 40–59 and ≥ 60 mm, but higher in C. gobio <40 mm L_T. Prevalence and mean intensity of infection over the 3 months did not differ either for the two sites individually or the pooled data. Mean mass of both uninfected and infected fish increased over this period, and were both highest in August as was the mean worm burden. There was, however, no difference in Fulton’s condition factor at any time. Mean mass of plerocercoids in C. gobio of ≥40 mm L_T increased significantly comparing June and July with August. Comparison of backcalculated L_T of infected and non‐infected bullheads of different age‐groups did not show any significant differences in growth at any age. In each parasite infrapopulation as the number of worms increased so their mean mass decreased significantly but at the same time the total mass of worms in the fish host remained more or less constant in relation to length. Aggregation of parasites in the fish population was low, as the majority were infected by one S. cotti plerocercoid. Gonad development was suppressed in infected C. gobio.     

Distribution of Cottus poecilopus Heckel and C. gobio L (Pisces) in Scandinavia

Andreasson. S. (Department of Animal Ecology, Lund University, Lund. Sweden.) Distribution of Cottus poecilopus Heckel and C. gobio L. (Pisces) in Scandinavia. Zool. Scripta 1(2): 69–78, 1972. –The distribution of the freshwater sculpins Cottus poecilopus Heckel and C. gobio L. in Scandinavia is presented. In southern Scandinavia there are rather defined limits between the species with C. poecilopus in the upper parts of the water courses and C. gobio in the lower reaches. There are few cases of sympatry. In northern Scandinavia there is a broad overlap and coexistence is common in the large rivers. The distribution pattern seems to be partly a result of an interaction between the species. In the South, the sharp limits are set by falls blocking the upstream dispersal of C. gobio thus indicating an exclusion of C. poecilopus in the lower reaches below the falls by the presence of C. gobio there. Where the species meet or overlap there is a habitat segregation.     

Uncoupling strong predator–prey interactions in streams: the role of marginal macrophytes

Predation refugia can facilitate the coexistence of predators and prey within an ecosystem by weakening trophic links between the two. The marginal macrophytes of shallow lakes are used facultatively by zooplankton to escape fish predation, leading to the stabilisation of lentic food webs. Little is known about such a role for the marginal macrophytes of lotic systems. In this paper, we examine whether the marginal macrophytes of chalk streams help buffer the interaction between the freshwater shrimp, Gammarus pulex and a benthic fish predator, Cottus gobio, both characteristically abundant in such systems. Quantitative surveys were taken of Gammarus and bullheads in winter and summer in a southern English chalk stream. These indicated that Gammarus seasonally switched their habitat preference, from marginal macrophytes in summer, to midchannel habitats in winter. Bullheads exhibited an opposite trend, preferring midchannel habitats in summer and all habitats, particularly marginal macrophytes, in winter. Large Gammarus and precopula pairs were found almost exclusively in the margins in summer. This spatial separation between Gammarus and bullheads in summer, though not in winter, was reflected in bullhead diets, as determined by gut analysis. In field manipulation experiments, bullhead presence was the strongest factor explaining the between‐habitat distribution of Gammarus in both summer and winter, indicating that the habitat shift of Gammarus was driven by the distribution of fish. Other abundant invertebrate taxa, including Asellus, mayflies and chironomids, exhibited little avoidance of habitat patches of high bullhead density. We conclude that marginal macrophytes in chalk streams can potentially facilitate the coexistence of high densities of both Gammarus and bullheads by spatially separating predator and prey in summer. They may further allow large populations of Gammarus to persist in the presence of high bullhead density at stretch‐wide spatial scales, by reducing the predation by bullheads of large breeding and newly born individuals of Gammarus in summer.     

Effects of temperature on persistence times of native and invasive gammarid species in the stomachs of Cottus gobio

Gastric lavage was used to investigate the effects of temperature on persistence time of two amphipod species, one native Gammarus pulex and one invasive Gammarus roeseli , in the stomachs of bullhead Cottus gobio . Persistence time was strongly influenced by temperature and prey type, such that G. pulex species degraded faster than G. roeseli .     

Identification of natural hybrids between Cottus poecilopus,Heckel, 1837, and Cottus gobio,Linnaeus, 1758, at a hybrid zone on the Svratka River (Czech Republic)

Sculpins occasionally form interspecific hybrids in areas of mutual contact. These can be verified genetically and, potentially, morphologically. At the study site (Svratka River, Czech Republic), Cottus gobio are common whereas C. poecilopus represent a geographically isolated alien population. Potential hybrids between these two species were compared with the parent populations to establish simple identification criteria. Analysis of nuclear and mitochondrial DNA confirmed the existence of a new European hybridisation zone. Hybrids were the result of unidirectional crossing, with female C. gobio taking the maternal position. With the exception of the lateral line position, morphometric characteristics and the epidermis structure were similar in many traits, reflecting the contribution of both species, as well as in the number of sacciform cells where hybrids were similar to C. poecilopus. The vertical position of the lateral line allows recognition of hybrids directly in the field, a useful feature for future studies. The occurrence of these hybrids confirms expansion of non‐native C. poecilopus at the expense of native C. gobio in rivers under anthropogenic pressure.     

Direct cloning of microsatellite loci from Cottus gobio through a simplified enrichment procedure

Applying a simplified enrichment procedure, we have isolated and characterized 177 microsatellite markers for Cottus gobio L., 1758 (Cottidae, Scorpaeniformes, Teleostei). In contrast to using specific probes for the enrichment, we use genomic DNAs of unrelated organisms for cross‐hybridization. This takes advantage of the fact that simple sequences are the only repetitive elements that are abundantly found in all eukaryotic genomes and that any genome usually contains all permutations of microsatellite motifs. This cross‐hybridization principle was employed to enrich genomic libraries of Cottus DNA to obtain a large number of nonredundant microsatellite markers without further screening procedures.     

Factors influencing the spatial distribution patterns of the bullhead (<Emphasis Type="Italic">Cottus gobio</Emphasis> L., Teleostei Cottidae): a multi-scale study

We used general linear modelling to assess the influence of environmental variables on the spatial distribution patterns of the bullhead (Cottus gobio) at stream system, site, and microhabitat scales in southwestern France. Bullheads occurred at 67 sites (out of 554 sampling sites), chiefly close to the source, in small and shallow streams. Population density at a site was primarily influenced by thermal conditions. Stream width was negatively related to the probability of presence of bullheads within the stream system, but positively related to local density, showing that bullhead density could increase within a range of stream width, but that wider rivers were unsuitable. Slope was negatively correlated to bullhead’s occurrence and local density, and depth was negatively correlated to local density and microhabitat use, suggesting that bullhead’s shimming performance was weak under greater erosive forces. Therefore, the most significant results suggested that the distribution of populations and individuals was first governed by the suitability of physical and hydraulic habitat, then population dynamics at a site was mainly governed by the thermal regime. Multi-scale studies of factors influencing a species’ distribution thus allow to integrate patterns observed at different scales, and enhance our understanding of interactions between animals and their environment. Such models are essential in the exploratory phase of fundamental and applied investigations, because they help to target further research, and they should influence the measures to be taken in field surveys or conservation plans.     

Effects of impoundment on populations of bullhead Cottus gobio L. and minnow, Phoxinus phoxinus (L.), in the basin of Cow Green Reservoir

The distribution, growth and reproduction of bullhead, Cottus gobio , L., and minnow, Phoxinus phoxinus (L.), were studied at Cow Green Reservoir, Upper Teesdale, for 10 years following impoundment in 1970. Comparisons were made with pre-impoundment results. The length-for-age of bullheads increased after impoundment. The length-for-age of minnows decreased. In winter both species were confined mainly to deep water or to the stone facing of the earth dam. In summer they were found throughout the reservoir though bullheads were scarce in the littoral zone. Female bullheads became sexually mature at an earlier age (84% at age-group I) than before impoundment (33% at age-group I) but no change was apparent amongst males (67–62% in age-group I). The post-impoundment sex ratio did not differ from 1:1 and individual fecundity did not change after impoundment. There was a significant reduction in individual fecundity of the minnow. The mean instantaneous rate of mortality ( Z ) for bullhead of age-groups II to IV was 0.96 ± 0.34 year⁻¹, compared with 0.85 ± 0.29 year in the Tees before impoundment.     

Competitive interactions for food resources between invasive racer goby Babka gymnotrachelus and native European bullhead Cottus gobio

Racer goby is one of several Ponto–Caspian gobiids spreading throughout European rivers and concurrent with recent declines in threatened populations of a native species of similar biology, the European bullhead. Although suggestive of competitive interactions, evidence thereof is scarce, so we examined behavioural interactions between racer goby and bullhead (single specimens of each species together, also pairs of each species) under experimental conditions (shared space with two shelters) to determine whether the invader displaces the native species when food resources are limited. Food (live chironomids) was added to a single feeder at rates below satiation levels twice over 24 h (once in light and once in darkness), with fish behaviour (aggressive interactions: attacks and threatening) and feeding activity (time spent near or inside the feeder) recorded using video cameras and infrared illumination. Racer goby exhibited aggressive behaviour towards bullhead (mean = 2.5 aggressive events h^?1), but rarely the inverse (threatening only, mean = 0.05 events h^?1), significantly limiting bullhead foraging time (by 62 %) and being faster to reach food in the feeding time in 76 % of cases. Gobies were more aggressive during daylight (77 % of all aggressive events occurring in light), and both species spent more time on feeding activities in darkness (88 and 66 % of all time spent in the feeder by bullheads and gobies, respectively). However, the adverse impact of goby on bullhead was independent of light conditions. Our results suggest that under natural conditions, racer goby are likely to displace bullhead during feeding, with potential consequences for foraging efficiency.     

Phylogeography of the bullhead Cottus gobio (Pisces: Teleostei: Cottidae) suggests a pre-pleistocene origin of the major central European populations

The bullhead Cottus gobio is a small, bottom-dwelling fish consisting of populations that have not been subject to transplantations or artificial stocking. It is therefore an ideal model species for studying the colonization history of central European freshwater systems, in particular with respect to the possible influences of the Pleistocene glaciation cycles. We sampled Cottus populations across most of its distribution range, with a special emphasis on southern Germany where the major European drainage systems are in closest contact. Mitochondrial D-loop sequencing of more than 400 specimens and phylogenetic network analysis allowed us to draw a detailed picture of the colonization of Europe by C. gobio. Moreover, the molecular distances between the haplotypes enabled us to infer an approximate time frame for the origin of the various populations. The founder population of C. gobio stems apparently from the Paratethys and invaded Europe in the Pliocene. From there, the first colonization into central Europe occurred via the ancient lower Danube, with a separate colonization of the eastern European territories. During the late Pliocene, one of the central European populations must have reached the North Sea in a second step after which it then started to colonize the Atlantic drainages via coastal lines. Accordingly, we found very distinct populations in the upper and lower Rhine, which can be explained by the fact that the lower Rhine was disconnected from the upper Rhine until approximately 1 million years ago (Ma). More closely related, but still distinct, populations were found in the Elbe, the Main and the upper Danube, all presumably of Pleistocene origin. Intriguingly, they have largely maintained their population identity, despite the strong disturbance caused by the glaciation cycles in these areas. On the other hand, a mixing of populations during postglacial recolonization could be detected in the lower Rhine and its tributaries. However, the general pattern that emerges from our analysis suggests that the glaciation cycles did not have a major impact on the general population structure of C. gobio in central Europe.     

River fragmentation increases localized population genetic structure and enhances asymmetry of dispersal in bullhead (Cottus gobio)

Man-made habitat fragmentation is a major concern in river ecology and is expected to have particularly detrimental effects on aquatic species with limited dispersal abilities, like the bullhead (Cottus gobio). We used ten microsatellite markers to investigate small-scale patterns of gene flow, current dispersal and neutral genetic diversity in a morphologically diverse river where fragmented and unfragmented sections could be compared. We found high genetic differentiation between sampling sites with a maximum F _ST of 0.32 between sites separated by only 35 km. A significant increase of genetic differentiation with geographical distance was observed in the continuous river section as well as in the full dataset which included headwater populations isolated by anthropogenic barriers. Several lines of evidence are consistent with the hypothesis that such barriers completely block upstream movement while downstream dispersal may be little affected. In the unfragmented habitat, dispersal rates were also higher in the direction of water flow than against it. The resulting asymmetry in gene flow likely contributes to the decrease of genetic variation observed from the lower reaches towards the headwaters, which is particularly pronounced in physically isolated populations. Our findings suggest that headwater populations, due to their isolation and low genetic variation, may be particularly vulnerable to extinction.     

Colonization history of the Swiss Rhine basin by the bullhead (Cottus gobio): inference under a Bayesian spatially explicit framework

The present distribution of freshwater fish in the Alpine region has been strongly affected by colonization events occurring after the last glacial maximum (LGM), some 20 000 years ago. We use here a spatially explicit simulation framework to model and better understand their colonization dynamics in the Swiss Rhine basin. This approach is applied to the European bullhead ( Cottus gobio ), which is an ideal model organism to study fish past demographic processes since it has not been managed by humans. The molecular diversity of eight sampled populations is simulated and compared to observed data at six microsatellite loci under an approximate Bayesian computation framework to estimate the parameters of the colonization process. Our demographic estimates fit well with current knowledge about the biology of this species, but they suggest that the Swiss Rhine basin was colonized very recently, after the Younger Dryas some 6600 years ago. We discuss the implication of this result, as well as the strengths and limits of the spatially explicit approach coupled to the approximate Bayesian computation framework.     

Microhabitat use and preferences of the endangered Cottus gobio in the River Voer, Belgium

Microhabitat use and preferences of juvenile and adult bullhead Cottus gobio , from the River Voer, Flanders, were studied and compared across different seasons. Water depth, water velocity near the substratum, surface water velocity and substratum type used by C. gobio differed between seasons. These differences, however, were not attributable to differences in microhabitat availability. Adults appeared to prefer higher water velocities and coarser substrata than the average ones available in the basin. Although water depth appeared to have little influence on seasonal variation of microhabitat use in adult C. gobio , juveniles preferred deeper water and coarser substrata in winter, whereas in summer they appeared to use shallower water. There was a difference in microhabitat use between juvenile and adult bullhead only in summer.     

The Alps as barrier to dispersal in cold-adapted freshwater fishes? Phylogeographic history and taxonomic status of the bullhead in the Adriatic freshwater drainage

The freshwater faunas of the Italian peninsula are isolated from the rest of Europe by the geographic barrier of the Alps and consequently have developed many endemic forms and contain few non-endemic species. However, some 'non-endemics' may either represent recent invaders of the Adriatic basin or cryptic endemic species. To test these two hypotheses against each other, we studied the origin and phylogenetic relationships of bullheads, cold adapted freshwater fishes of the genus Cottus, from both sides of the Alps and Dinaric Mountains. From the Adriatic basin, Cottus ferrugineus () was described as an endemic species, but the present analyses of sequences of the complete mitochondrial control region of 146 individuals from 43 localities showed no major differentiation between bullheads from both sides of the Alps. The very low diversification between representatives across the Alps suggests active transfers of haplotypes across this geographic barrier from the glacial cycles up to recent times. The transfers are most likely based on stream capture, since the cold-adapted bullhead is able to colonise the highest stretches of the water courses. No other freshwater fish in Europe is known to have experienced such an extensive gene flow across the highest European Mountains. In contrast, the Dinaric Mountains seem to have been a much more effective barrier between the Danube and the Adriatics. Our data reject the hypothesis of C. ferrugineus as an endemic species in the whole Adriatic drainage.     

Comparative life histories and microhabitat use in three sympatric sculpins (Cottidae:Cottus) in northeastern California

Synopsis Life histories of three sculpins endemic to the Pit River system, northeastern California, are detailed.Cottus pitensis is widely distributed throughout the drainage. This species is typically found in rapidly flowing, shallow water on coarse substrate. It grows to a large size, >100 mm, and lives to 5 years. Spawning occurs annually, in early spring. Fecundity is relatively low, <320 oocytes.Cottus asperrimus is restricted to a large population in Fall River and smaller concentrations in Hat Creek and Pit River. It is a relatively small fish, <80 mm, and lives to 5 years. It occurs in deep, slow-moving, often spring-fed streams, typically on fine substrates. Spawning lasts from autumn to early spring. Males establish nests on hard substrates and guard several clutches of eggs. Fecundity ranges from 140–580 oocytes.Cottus klamathensis macrops is relatively rare in the drainage. The largest concentration is in the Hat Creek system, smaller populations exist in Fall River and Pit River. This sculpin is found on coarse substrate, often where aquatic vegetation is abundant. It is typically found in slow, relatively shallow water. It grows to >100mm and lives to 5 years. Males guard eggs during the short spawning season beginning in late winter. Fecundity in this species reaches a maximum of 650 oocytes. Life history strategies vary within the genus, yet the life histories of these sculpins are more similar to each other than they are to many of their congeners. This within-group similarity may have resulted from exposure of all three species to similar environmental pressures. This is predicted by life-history theory.