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.     

Conservation units based on mitochondrial and nuclear DNA variation among European bullhead populations (Cottus gobio L., 1758) from Flanders, Belgium

In this study, we aimed to delineateevolutionarily significant units (ESUs) andmanagement units (MUs) for the Europeanbullhead in Flanders (Belgium). Therefore, wedetermined the genetic interrelationshipsbetween 11 bullhead populations, using lengthvariation at 7 polymorphic microsatellite lociand sequence variation in the d-loop of themitochondrial DNA (mtDNA). Despite therelatively small geographical scale of ourstudy, the analysis of the d-loop sequencesshows that the Flemish bullhead populationscontain 3 haplotype groups, which can beassigned to 3 previously described EuropeanmtDNA clades. Because of the importantdifferences between these clades, they may bedefined as evolutionarily significant units,which should be managed separately. Analysis ofmicrosatellite data reveals very high degreesof isolation between populations, with theexception of 3 pairwise comparisons whichinvolved adjacent populations. Our data suggestthat the 3 haplotype groups probably qualify asESUs, as they show phylogeographicdifferentiation for mtDNA variants as well assignificant divergence of allele frequencies atnuclear loci. However, one of these units,limited to a single population, may be ofCentral European origin. All populations of theScheldt basin meet the criteria for MUrecognition, since significantly differentmicrosatellite allele frequencies as well asprivate alleles are found. In contrast, geneticdifferentiation among the 3 populations of theMeuse basin is very low.     

Histological and enzyme histochemical changes in the kidney of male bullhead (Cottus gobio) during the spawning period

Histological and enzyme histochemical studies were carried out on the excretory kidney of the male bullhead ( Cottus gobio ). During the spawning season striking morphofunctional changes were observed in the second proximal segment of the kidney tubule. The tubular epithelium was greatly hypertrophied, strongly basophilic and produced a PAS-positive secretion. The enzyme histochemical pattern also changed conspicuously during this time: the alkaline phosphatase activity in the brush border was greatly reduced; the acid phosphatase and non-specific esterase activity in the cytoplasm was distinctly elevated.     

Genetic variability in the European minnow,Phoxinus phoxinus (L.)

An electrophoretic study of genetic variation at 11 loci was performedfor a population of European minnows, Phoxinus phoxinus (L.). Ten loci, EST-1 ^*, EST-2 ^* EST-3 ^*,GPD-1 ^*,GPD-2 ^*,GPI-1 ^*,GPI-2 ^*,MPI ^*,6PGD ^* and PGM ^* were polymorphic. IDH ^*wasmonomorphic. The mean number of heterozygotic loci over all 176 fish was 3.05 ± 0.104(SE). Observed mean heterozygosity was 0.28±0.058(SE) and expected mean heterozygosity was 0.27±0.054(SE). EST-2 ^*, EST-3 ^* andPGM ^* were not in Hardy-Weinberg equilibrium. Length,condition, parasite numbers or male breeding characters, i.e. red colorationand tubercles, were not influenced by single enzyme loci.     

The feeding habits of 0+ stone loach, Noemacheilus bavbatulus (L.), and bullhead, Cottus gobio L.

The feeding habits of the 0+ year class of two of the most common small fish species in the River Ouzel, Milton Keynes, stone loach, Noernacheilus bnrhafulus (L.), and bullhead, Coftus gobio L., are assessed throughout the first growing season and compared. Despite similarities in the diets of the two species, differences in feeding behaviour and in morphology, especially in gape, may serve to reduce interspecific competition by partitioning of food resources.     

Population biology of the helminth parasites of Phoxinus phoxinus (L.), the minnow, in a Cardiganshire lake

Specimens of Phoxinus phoxinus (L.) were collected over a period of a year from September 1967 to September 1968 from Frongoch Lake, Cardiganshire, Wales. Of a total of 650 fish examined for helminth parasites, 32 % were definitive hosts for Macrolecithus papilliger , and 16·5% for Neoechinorhynchus rutili (Müller), both parasites of the alimentary canal. All the fish examined were second intermediate host for Diplostomum phoxini (Faust) in the brain and 1·67% for Ligula intestinalis (L.) in the coelom. An increase in incidence of infestation with M. papilliger in spring and autumn and with N. rutili in summer were evident. The sex of the fish had no apparent effect on worm burden whereas size was more significant. There was a decrease in incidence of infestation with M. papilliger and an increase with N. rutili together with an increase in intensity of infestation with D. phoxini as the fish became larger. The latter species accumulated in the brain as the host increased in age.     

The problem of sexual inversion in the minnow, Phoxinus phoxinus (L.)

The incidence of sexual inversion in Phoxinus phoxinus has been studied by examining histologically (a) the gonads of 406 adult specimens caught from the wild during a period of 16 months, (b) the gonads of 168 adults kept in captivity in groups with an experimentally altered sex-ratio, and (c) the differentiation of the gonads in the fry.
All the adults had either testes or ovaries and no case of intersexuality was observed. For both testis and ovary, we recognized two phases, depending on the seasonal cycle: an active spring-summer phase, corresponding with the period of reproduction, and a quiescent autumn-winter phase. The ovaries showed an asynchrony in oocyte development, which is typical of species that spawn many times during the breeding season.
In P. phoxinus the ovaries are easily recognizable just one month after hatching and testes are well differentiated in 2-month-old fingerlings. Differentiation is, therefore, precocious and follows a pattern typical of gonochoristic species in which hermaphroditism never occurs.     

Grouping fails to affect the growth and energy budget of a cyprinid, Phoxinus phoxinus (L.)

The effects of grouping on the growth and some components of the energy budget of the minnow, Phoxinus phoxinus (L.), were tested in the laboratory. No effects of grouping were detected on absorption efficiency, the proportion of food energy lost in nitrogenous excretion, maximum rate of food consumption and conversion efficiency. Grouping had no significant effect on either specific growth rate or the dry matter content of individual fish.     

The life history of the minnow Phoxinus phoxinus (L.) in a productive stream

SUMMARY. 1. In the River Frome, Dorset, this small cyprinid spawned from mid-April to early August and consequently some 0 group minnows did not develop scales until the following spring.
2. Most I group fish formed a summer hyaline band on their otoliths in a edition to the normal winter band.
3. Life-span was short with a winter age-structure of 67% 0 group, 32% I group and 1% II group fish with mean fork lengths of 29, 55 and 73 mm respectively.
4. In June, substantial numbers of I group fish reached reproductive size (50mm) and thereafter progressively replaced II group fish in the spawning shoals. Few II group males survived beyond June, few females beyond August.
5. Only half of I group minnows reached matutity. These were usually fish which had reached 30mm by the previous winter. Very few of these large I group fish survived to spawn the following year.
6. This differential mortality, together with the loss of condition and disappearance of II group fish during spawning, may indicate the cost of an extended high level of reproductive effort.
7. In this productive environment the life history of the minnow more closely resembles that of the small sympatric species Cottus gobio and Noemacheilus harhatulus than it does that of sympatric cyprinids.     

Formation of parasite fauna in the bullhead (Cottus gobio L.) (Pisces: Cottidae)

The formation of parasite fauna in the bullhead Cottus gobio L. in different water bodies was examined. The largest number of parasite species including specific parasites was observed in the Onega and Ladoga lakes. It is suggested that the host and their parasites migrated namely from these water bodies to other part of the range of the bullhead.     

Gyrodactylus onegensis sp. n. (Monogenea)--a parasite of the bullhead (Cottus gobio L.)

A new species, Gyrodactylus onegensis sp. n., is described from gills of the freshwater sculpin Cottus gobio.     

The effects of spinal nerve section on responses of melanophores in the minnow, Phoxinus phoxinus(L.)

A continuous observation apparatus was used to study the responses of Phoxinus phoxinus melanophores to illuminated black/white backgrounds and their reversal. The fish. Although confined, showed maximum melanosome dispersion (MI 5) and maximum melanosome aggregation (MI 1) when exposed to illuminated black and white backgrounds respectively. Melanophores affected by spinal nerve section showed full melanosome dispersion and the affected area appeared as a black band. The affected melanophores marginally and gradually aggregated their melanosomes if the fish was exposed to an illuminated white background for about a week. The responses of these melanophores to illuminated black and white backgrounds and their reversal indicates that the dispersal of their melanosomes in response to a black background is much faster than their aggregation in response to a white background. It is concluded that an active mechanism is involved and possible factors controlling it are discussed.     

Accumulation and effects of aluminium in the minnow (Phoxinus phoxinus L.) at different pH levels

The accumulation and effects of 150 pg All^-1 on minnows ( Phoxinus phoxinus L.) were studied in soft water at pH 7, 6 and 5. The fish were kept in a flow-through water system for up to 48 days. Addition of aluminium to the water resulted in poor appetite, passivity and a tendency for the fish to gather as far as possible from the aluminium inlet. The mortality was high at pH 5 and also after addition of aluminium at pH 6. Severe lesions were noted in the gills and olfactory organs after exposure to aluminium. The concentration of aluminium in the gills, liver and kidney was increased at low pH. A large proportion of the aluminium was located on the surface of the gill epithelium. The recovery capacity was pronounced when the aluminium exposure was terminated and the pH was increased from 5 to 7. No further mortality occurred and gill and olfactory organ structure returned to near normality in about 36 days.     

Preliminary observations on different reproduction strategies in the bullhead (Cottus gobio L.) in northern and southern England

The size distribution and numbers of oocytes were examined in samples of ovaries from two sites and in fish in tanks whose past spawning activities were known. In an upland stream in northern England Cottus gobio would seem to be an annual spawner, maturing a single batch of eggs each year. In a more productive stream in southern England, where the reproductive life is short, fish mature and lay a succession of batches of eggs during a long spawning season. The physiological and ecological implications of these characteristics are discussed.     

Host age and structure of the component communities of parasites in river minnow Phoxinus phoxinus (L.)

Species composition and structure of the communities of fish parasites in river minnow Phoxinus phoxinus (L.) from the Pechora river were investigated in two of the Pechora-Ilychsky Biosphere Nature Reserve, Komi Republic. The component communities of the parasites in river minnow are shown to have a one-year cycle including the states of development, completion, and destruction. Communities in the state of development are characterized by a low variety of species, low values of Shannon index, often high values of domination index, presence of only two groups of parasites in the structure described by variational curved of the conditional biomasses of species, deviation of the conditional biomasses of species from the linear regression, and sum of errors of the regression equations lower a threshold value. The communities consist of young individual parasites and their larval stages. Completed community is characterized by the following properties. There are three groups of parasites, differing in allometric index, in the structure, discerned by the ratio of conditional biomasses of the species included. Conditional biomasses of species in ecologically safe reservoirs lie on the segments of straight lines. Species variety reaches its maximum. Species are presented mainly by mature specimens and larval stages of the parasites using fish as intermediate host. Community in the state of destruction shows low values of domination index and relatively small variety pf species. Such community is consist of one or two groups of species, which are represented by mature, oviparous, and dying individuals. There are larval stages of parasites using fish as intermediate hosts. Dominant species or species groups, as well as values of indexes describing the component communities of parasites, can be different in mature river minnow from different geographical regions. However, the number of groups of parasites, formed by the ratio of conditional biomasses, remains constant, and sum of errors of the regression equations characterizing the communities is always below 0.25. Component communities of parasites in young river minnows differ from the communities in mature fishes by lower variety of species. Lower biomass, and lower number of individual parasites. The communities of parasites in 0+ old fishes are often characterized by lesser number of groups of parasites, classified by the ratio of their biomasses, and presence of two dominate species. It is often impossible to count the sum of errors of the regression equations describing spread in values of biomasses of the species forming the community.     

Component parasite communities in the minnow Phoxinus phoxinus (L.) from ecologically safe and polluted reservoirs

A comparative analysis of the structure of component parasite communities in the minnow from ecologically safe or polluted to variable extent reservoirs was made. The analysis showed that the increase of the reservoir pollution by everyday drains, washings of fertilizers from the fields around and the dacha sections causes the change of dominant parasite species in the minnow, alteration of the sum of errors in the regression equation characterizing the spread of values of biomasses of the species forming the parasite community, and change of the "graphic" structure of the community. It is proved, that the high concentration of biogens has a destructive effect on natural systems, and at the same time causes the process of self-organizing, leading to the alteration of the community structure. As soon as even a small part of biogens is stopped to come into the reservoir, and the quality of environment is restored, the community restores its structure.     

The adenohypophysis of the flounder,Pleuronectes flesus,and the minnow,Phoxinus phoxinus

Summary The pituitary gland of the flounder, Pleuronectes flesus, showed several unusual cytological features. Between the RPD and the PPD was a zone of cells that stained purple with Alcian blue—PAS—orange G. Many of these cells were apparently degenerating. In the PPD the strands and coils of presumptive STH cells showed a tremendous variation in both size and staining properties. In the PI there were two cell types, the PAS-positive one bordering the neurohypophysis. Around the periphery of the PI was a zone of chromophobic cells, and throughout the PI were numerous intracellular and extracellular acidophil spheres.No well defined ACTH cells were found in the RPD of the minnow (Phoxinus phoxinus). The Alcian blue—PAS—orange G technique distinguishes between blue TSH cells and purple GTH cells in the RPD and PPD. GTH cells from animals collected in the winter were vacuolated. The PI contained two cell types whose staining reactions and ultrastructure were extremely variable. Intra- and extra-cellular acidophil spheres were present.I should like to thank Dr. T. Kerr of the Zoology Department, Leeds University, for his help and encouragement, and Mr. J. Dingley of the Department of Zoology, Aberystwyth, for collecting the flounders.     

The effect of extreme northerly climatic conditions on the life history of the minnow, Phoxinus phoxinus (L.)

Over its exceptionally wide geographical range the minnow displays a remarkable variability in its life history. In southern England many fish mature after one year and few fish survive to their third birthday. Very few fish spawned for more than one season but a 65-mm female is estimated to spawn up to 3172 eggs over the extended spawning season. In the River Utsjoki in Finnish Lapland maturity was strongly size-dependent and delayed until the fish reached 5, 6 or even 7 years of age, with a maximum age of 13 years at a length of only 75 mm. Unlike some other small-sized species in unproductive environments, individual females continued to spawn successive clutches of eggs though over a much shorter spawning season. The maximum estimated egg production was 824 for a 65-mm fish. Temperature had a strong direct or indirect effect on growth, with significantly lower growth increments in cold summers. At a second Arctic site in eastern Lapland growth rates were higher and maturity earlier, yet clutch size was greatly reduced with a seasonal fecundity estimate of only 314. There were only small differences in size of eggs or larvae between the populations. Evidence from other species indicates that most of the observed changes over the range of the minnow are phenotypic responses to the contrasting environments. However, the different strategies displayed at the two Arctic sites could represent the results of selection for differences in pre- and post-reproductive survival.