Danish Localities Research Articles

Cortinarius koldingensis—a new species of Cortinarius, subgenus Phlegmacium related to Cortinarius sulfurinus

Cortinarius koldingensis is described as a new species based on several collections from a small area in Denmark. Sequence data from the ITS marker places it in the Sulfurini group of the large Calochroi clade of Cortinarius subgenus Phlegmacium, along with C. sulfurinus and two North American taxa. It is phylogenetically distinct from the other species in the Sulfurini and differs from C. sulfurinus by several deviating morphological characteristics. The species has been collected on several occasions over the last 15 years from only one Danish locality. Despite extensive molecular screening of numerous possible conspecific specimens, including holotype and comparison with all publicly available ITS sequence data from specimens and environmental DNA alike, no match with specimens from elsewhere has been encountered, and we hypothesize that the species is rare in Europe.

Monitoring the near-extinct European weather loach in Denmark based on environmental DNA from water samples

The European weather loach (Misgurnus fossilis) represents one of many European freshwater fishes in decline. Efficient monitoring is essential if conservation efforts are to be successful, but due to the species’ cryptic biology, traditional monitoring methods currently in use are inefficient, time consuming and likely prone to non-detection error. Here, we investigate the usefulness of environmental DNA (eDNA) monitoring as an alternative or supplementary method for surveying the Danish weather loach population, which is presumed to consist primarily of a single group of no more than 50 individuals. In 2008, the majority of historical Danish localities were surveyed, using traditional fishing techniques. We then applied eDNA methods to a number of these, as well as other potential localities. We successfully detected the weather loach at multiple sites in the single known remaining locality; a result that was later confirmed when local managers caught eight live specimens. Furthermore, the eDNA method indicated presence of the weather loach in another historical locality, where the species has not been observed since 1995. At the remaining localities, weather loach eDNA was not detected, providing further evidence for its absence. Importantly, the eDNA survey required less effort in person-hours and lower costs than the traditional fishing survey. This study confirms that eDNA monitoring is a valid supplement to traditional monitoring methods currently applied to monitor rare freshwater fishes. We propose that by providing reliable distribution data at lower cost and limited effort, the eDNA method can allow for increased management efficiency of endangered freshwater species such as the European weather loach.

Genetic diversity in the parthenogenetic reproducing tardigrade Echiniscus testudo (Heterotardigrada: Echiniscoidea)

Little is known about the genetic structure of microscopic animals from mosses and lichens. A few studies have investigated the geographic variation in tardigrades from mosses, but so far no study has investigated the intra-population or local clonal lineage variation. Echiniscus testudo (Echiniscoidea: Echiniscidae) belongs to a large cosmopolitan genus of terrestrial tardigrades comprising more than 150 species. It is a common tardigrade in mosses in the temperate part of the Northern hemisphere, and is highly tolerant of desiccation and freezing. In a previous study, we reported a maximum of 1.28% sequence variation (uncorrected p-distance) in cytochrome c oxidase subunit I (COI) haplotypes between clonal lineages covering a large geographical area. However, in this previous study we used pooled specimens to constitute a sample, and the genetic diversity from single specimens within a locality therefore remains unknown. Accordingly, the present study investigates the COI sequence variation and haplotype diversity between single specimens of E. testudo collected at three Danish localities, separated by 80 m and 186 km. A total of 10 COI haplotypes were found in the present study (Et2, Et3, Et9, Et12-Et18); only three of these were previously reported (Et2, Et3 and Et9). The uncorrected COI sequence diversity ranged between 0-2.07%, with haplotype Et18 having the highest genetic difference. The second most variable haplotypes (Et14, Et15, and Et17) all showed a maximum diversity of 1.19% compared to the other haplotypes. No general pattern of haplotype distribution was evident. Our data suggest that E. testudo has dispersed across the Baltic sea as haplotypes Et3, Et13 and Et14 are present at all three localities. The most likely dispersal mode is passive wind dispersal in the cryptobiotic tun stage. The current study emphasises that numerous sequences from single specimens are needed to describe the genetic diversity within single moss cushions.

Biomonitoring with lichens on twigs

AbstractTwo surveys of the lichen and bryophyte flora growing on oak twigs from a Welsh and a Danish locality were compared with additional data on bark pH and % nitrogen in thalli ofHypogymnia physodes. Despite differences in climate and lichen flora, both sites showed a shift in the lichen communities from nitrogen sensitive (nitrophobe) to nitrogen tolerant (nitrophile) species, which was correlated with both increasing bark pH and an increase in total nitrogen in thalli ofH. physodes. The floristic survey from Wales was a repetition of a study eight years earlier (Wolseley & Pryor 1999) now showing a loss of nitrophobes in all sites and the appearance of nitrophiles in pasture sites in 2003. This study demonstrates that lichens on twigs can be used as an early warning system to detect a response to changes in land management and nitrogen deposition.

First record of a chiton from the Palaeocene of Denmark (Polyplacophora: Leptochitonidae) and its phylogenetic affinities

Synopsis A new species of fossil polyplacophoran from the Danian (Lower Palaeocene) of Denmark is described from over 450 individual disarticulated plates. The polyplacophorans originate from the ‘nose‐chalk’ in the classical Danish locality of Fakse Quarry, an unconsolidated coral limestone in which aragonitic mollusc shells are preserved through transformation into calcite. In plate architecture and sculpture, the new Danish material is similar to Recent Leptochiton spp., but differs in its underdeveloped apophyses and high dorsal elevation (height/width ca. 0.54). Cladistic analysis of 55 original shell characters coded for more than 100 Recent and fossil species in the order Lepidopleurida shows very high resolution of interspecific relationships, but does not consistently recover traditional genera or subgenera. Inter‐relationships within the suborder Lepidopleurina are of particular interest as it is often considered the most ‘basal’ neoloricate lineage. In a local context, the presence of chitons in the faunal assemblage of Fakse contributes evidence of shallow depositional depth for at least some elements of this Palaeocene seabed, a well‐studied formation of azooxanthellic coral limestones. This new record for Denmark represents a well‐dated and ecologically well‐understood fossil chiton with potential value for understanding the radiation of the Neoloricata.

Genetic structure of the European polecat (Mustela putorius) and its implication for conservation strategies

AbstractDuring the last century, the European polecat Mustela putorius populations in most of Europe declined and survived in fragmented patches, because of habitat alterations and direct persecution. To assess the genetic consequences of the demographic decline and to describe the spatial pattern of genetic diversity, 250 polecats sampled at seven localities from five European countries – Poland, Denmark (southern Denmark and northern Denmark), Spain, Belgium (eastern and western) and the Netherlands – were screened by means of nine microsatellite loci. Genetic diversity estimated by mean expected heterozygosity (HE) and allelic richness (AR) were moderately high within populations [range: 0.50 (northern Denmark) ≤HE≤0.64 (Poland) and 1.33≤AR≤7.80] as compared with other carnivores and mustelids. Bottleneck tests suggested that polecat populations in southern Denmark and Poland have declined recently and populations from northern Denmark and the Netherlands have expanded recently, whereas the remaining populations did not show any sign of demographic change. Recent demographic changes could suggest that some of the populations are still not in equilibrium, which could partly explain the relatively high genetic variability observed in polecat populations despite the drastic decline in population size observed in several European countries. A significant heterozygote deficiency [FIS=0.19; 0.01≤95% confidence interval (CI)≤0.32] suggests substructuring within the total European sample. Partitioning of the genetic variation among sampling locations (FST=0.14; 0.06≤95% CI≤0.23) and pairwise FST between localities (range: 0.01≤FST≤0.37) without any correlation with the geographic distances between localities were found, suggesting a recent divergence and a restriction of gene flow between populations and the action of genetic drift. An assignment test showed that the Polish and the northern Danish populations were the most unique, whereas the other populations were partially admixed. Factorial component analysis tests indicate a subdivision of the total sample into two distinct groups: one including the samples from Poland and the two Danish localities and the second group comprising the remaining localities investigated. The observed pattern of genetic differentiation is suggested to be due to two main routes of recolonization after the last glacial period. To compare the results obtained with microsatellite data, the most variable region of the mitochondrial DNA (d‐loop) was sequenced and different phylogenetic reconstructions and genetic diversity analyses based on nucleotide (π) and haplotype diversity (h) measures within populations were performed using a subsample of populations. The lack of well‐defined geographical structure, as well as the reduced level of mitochondrial DNA variability (π: 0.00274±0.00038; h: 0.876±0.028) that was found, has been previously reported in several studies on different carnivores and supports the hypothesis of post‐glacial recolonization from southern or eastern refugees of Europe as suggested by the microsatellite data. Implications for conservation strategies of the polecat at the European level are discussed.

AFLP markers for the R-gene in the flea beetle, Phyllotreta nemorum, conferring resistance to defenses in Barbarea vulgaris

A so-called R-gene renders the yellow-striped flea beetle Phyllotreta nemorum L. (Coleoptera: Chrysomelidae: Alticinae) resistant to the defenses of the yellow rocket Barbarea vulgaris R.Br. (Brassicacea) and enables it to use it as a host plant in Denmark. In this study, genetic markers for an autosomal R-gene, inherited as a single, dominant locus in flea beetles from the Danish locality "Kvaerkeby" are described, and a genetic linkage map around this particular R-gene is constructed, using the technique of AFLP (Amplified Fragment Length Polymorphism).

Spirorchid and serpulid polychaetes are candidates as invertebrate hosts for Myxozoa

While, in recent years, it has been routine to elucidate the life cycles of freshwater myxozoans, which have oligochaetes and bryozoans as invertebrate alternate hosts (Kent M.L., Andree K.B., Bartholomew J.L., El-Matbouli M., Desser S.S., Devlin R.H., Feist S.W., Hedrick R.P., Hoffmann R.W., Khattra J., Hallett S.L., Lester R.J.G., Longshaw M., Palenzeula O., Siddall M.E., Xiao C. 2001: J. Eukaryot. Microbiol. 48: 395-413), nobody has yet unravelled the life cycle of any of the numerous species which occur in marine fish (Lom J., Dykova I. 1992: Protozoan Parasites of Fishes. Elsevier, Amsterdam, 315 pp). Even though actinosporeans have been found in marine oligochaetes (Hallett S.L., Lester R.J.G. 1999: Int. J. Parasitol. 29: 419-427; Hallett S.L., Erseus C., Lester R.J.G. 1999: Syst. Parasitol. 44: 49-57) it is expected that non-oligochaete invertebrates, especially polychaetes, may act as invertebrate hosts for several of the marine species of the Myxozoa. The only known examples of marine non-oligochaete, invertebrate myxozoan hosts are the sipunculid worm Nephasoma minuta (Keferstein) (syn.: Petalostoma minutum) from the Channel, England (Ikeda I. 1912: Arch. Protistenkd. 25: 240-272) and the polychaete Nereis diversicolor O.F. Muller (Nereidae) from a Danish locality (Koie 2000: J. Parasitol. 86: 871-872). Studies of marine polychaetes have shown that Spirorbis spirorbis (Linnaeus) (syn.: S. borealis Borg) (Spirorbidae) and Hydroides norvegica Gunnerus (Serpulidae) from Danish waters are infected with actinosporeans of unknown myxozoans. In the northern Oresund, Denmark, the regularly coiled, calcareous tubes of S. spirorbis mostly occur on algae of the genera Fucus and Laminaria, whereas the larger, irregularly coiled, calcareous tubes of H. norvegica mostly are attached to the shells of live scallops. Spirorbis spirorbis exhibits a restricted breeding season during the warm months, whereas H. norvegica apparently breeds throughout most of the year. Algae with S. spirorbis and bivalves with H. norvegica were dredged at depths of about 15 m and 26-28 m, respectively. The infections were not followed throughout the year, but S. spirorbis was found infected with actinosporean stages from March throughout the summer to September. Hydroides norvegica was found Fig. 1. Ventral view of a live flattened, slightly infected specimen of Spirorbis spirorbis removed from the calcareous tube. The pansporocysts occur in segments with the testes (t) posterior to the ovaries. The eggs (e), stomach (s) and contents of the intestine (i) appear dark in transmitted light. Scale bar = 0.5 mm.

Natural Coprevalence of Strongwellsea castrans, Cystosporogenes deliaradicae, and Bacillus thuringiensis in the Host, Delia radicum

Adult cabbage root flies (Delia radicum) from three Danish localities were diagnosed microscopically for the natural prevalence of Strongwellsea castrans, Cystosporogenes deliaradicae, and Bacillus thuringiensis. C. deliaradicae was significantly coprevalent with S. castrans. B. thuringiensis sporangia were diagnosed in the hemolymph in two D. radicum which were also infected with S. castrans and proved to belong to serovar aizawai and serovar balearica. The biological characterization of S. castrans proved that at 17.5°C flies developed an abdominal hole 7.9 days (mean) after infection and that 5.7 days (mean) passed from the emergence of the hole to the death of the infected host. No mortality effect among D. radicum subjected to B. thuringiensis serovar aizawai, balearica, and kurstaki isolates was detected. RAPD with DNA proved that six B. thuringiensis serovar balearica isolates (all from the same fly) were indistinguisable. This indicates that proliferation of B. thuringiensis in the abdomen of an S. castrans-infected D. radicum may be due to just one genotype. The profiles of one isolated aizawai strain did not correspond to the profiles of other serovar aizawai strains used for comparison. The biological significance of the interaction between the involved pathogens is discussed.

Gene Flow Among Populations of the Leafbeetle Species Gastrophysa polygoni and Lochmaea suturalis in Denmark (Coleoptera: Chrysomelidae)

The genetic differentiation of populations of the leaf beetles Gastrophysa polygoni Linnaeus 1758 and Lochmaea suturalis Thomson 1866 was measured by use of allozymes in order to estimate the gene flow between geographically separated populations. 264 specimens of G polygoni from 4 Danish localities and 286 specimens of L suturalis from 3 Danish localities were analysed for 12 and 9 enzymes, respectively. 11 out of 17 loci from G polygoni were polymorphic and 8 out of 10 from L suturalis. No difference in the allelic distribution between sexes within each population of G polygoni was found. The level of heterozygosity was of the same magnitude for all seven populations studied. For each species a high genetic identity between the populations was found, but lack of homogeneity between allelic distributions of the populations revealed local divergence. The estimated gene flow corresponded to 3-4 reproducing individuals per generation for each species, which indicates that genetic exchange between the populations does occur.

Effects of human disturbance on broods of redbreasted mergansers Mergus serrator: J. Kahlert, Wildfowl, 45, 1994, pp 222–231

The effects of human disturbance on broods of Red-breasted Mergansers were studied at three Danish localities. Fast-moving boats (fishermen, windsurfers, and motor boats) were considered to have the greatest impact on Merganser broods a frequency of encounters and rate of disturbance were highest. The presence of fast-moving boats also caused the most significant modifications to time budgets. However, Merganser broods are considered quite resilient to human disturbance as they quickly resumed their former activities after disturbance, and at moderate disturbance levels, interruptions to comfort behaviour were compensated fro within 1 hours. However, on one locality with high disturbance levels and frequent high-speed boating, the survival of ducklings was reduced.

Prevalence of antibodies to Borvelia burgdorferi in Danish rodents

Six rodent species from six Danish localities were examined for antibodies to Borrelia burgdorferi by immunofluorescence assay (IFA). A total of 1097 specimens were tested using a serum titre of 1:8. The wild mice Apodemus sylvaticus and A. flavicollis had high prevalences of B. burgdorferi antibodies of 42.1% and 27.9% respectively, but the vole Microtus agrestis also showed an exceptionally high prevalence of 32.7%. Clethrionomys glareolus had a low average prevalence of 17.4%. The lowest and highest prevalences of rodents seropositive for B. burgdorferi of 6.5% and 100% were found for Micromys minutus and Mus musculus respectively. Twice as many of the Microtidae voles (M. agrestis and C. glareolus) as of the family Muridae were caught, and in Denmark the actual number of seropositive Microtidae was 1.6 times higher than for Muridae. The results indicate that in Denmark the family Microtidae species plays an important role as an animal reservoir host for B. burgdorferi. The prevalence of antibodies against B. burgdorferi varied from locality to locality, but seropositive rodents were found at all localities examined, indicating that the spirochete was widely present in Denmark.

A revision of the taxonomic position of Syncrypta glomerifera (Chrysophyceae), establishment of a new genus Lepidochrysis and observations on the occurrence of L. glomerifera comb. nov. in brackish water

Syncrypta glomerifera, a species originally isolated and described from a salt marsh pool in England has been recorded from brackish water in several samples from the Baltic Sea along the Finnish coast, and also from a Danish locality. Both colonial and monad forms have been studied with LM and TEM. The systematic position of the species is discussed and the new genus Lepidochrysis gen. nov. is established.

Indicator studies: a critical review and a new data­presentation method

This paper deals with general problems of indicator collection and introduces a new datapresentation method, named the circle-map method. The advantages of this method is demonstrated by collecting a wide spectrum of Scandinavian and Baltic rock types in two Danish localities. It is concluded that an indicator count should be made in situ and comprise at least 50 stones, and that the rock types used should represent parent areas as evenly distributed as possible throughout the Scandinavian and Baltic regions. The circle maps indicate the routeway of ice flow from the Scandinavian mountains to the locality dealt with. It emphasizes variations in indicator content and may, therefore, show the lateral displacement of ice-flow routes which should be expected from a glaciological point of view. In addition, the method makes it possible to distinguish between directly brought stones and contaminating stones.

Gametic disequilibria between virulence genes in barley powdery mildew populations in relation to selection and recombination. II. Danish observations

Analyses of gametic disequilibria (linkage disequilibria) between virulence genes were carried out on 14 samples of Erysiphe graminis f.sp. hordei collected from the aerial population at six Danish localities from 1985 to 1988. In most samples, the virulence genotypes Va7 Va12, Va9 Va12, VarV(la). Vk V(la) and VhV(La) were significantly less frequent than the product of the corresponding gene frequencies (negative gametic disequilibrium), while the virulence genotypes VaVk,Va9VnVh, VvVn and VkVh and were significantly more frequent than the product of the corresponding gene frequencies (positive gametic disequilibrium). These results could largely be explained by the distribution of powdery mildew resistance genes in the barley varieties grown in Denmark. Selection forces induced by two resistance genes present mainly in different varieties were likely to generate negative gametic disequilibrium between the corresponding virulence genes, whereas selection forces induced by two resistance genes present mainly in the same variety were likely to generate positive gametic disequilibrium between the corresponding virulence genes. This pattern was in accordance with that predicted from two‐ and three‐locuS models comprising recombination and selection induced by host resistance genes (Øsstergård & Hovmøller, 1991). It was concluded that such a model system is necessary for designing an ideal strategy for the deployment of varieties (resistance genes), and that the signs of gametic disequilibria do not provide adequate information for that purpose.

Stable isotope and trace element stratigraphy across the Cretaceous/Tertiary boundary in Denmark

Carbon and oxygen isotope analyses and strontium measurements were made on Cretaceous/Tertiary (K/T) boundary samples (bulk rock, one benthonic foraminiferal species, and one planktonic foraminiferal species) from two Danish localities - Stevns Klint and Kjolby Gaard. Analyses of closely spaced samples across the K/T boundary reveal a large (2‰) positive oxygen isotope excursion coincident with the iridium anomaly in the Fish Clay, followed by large fluctuations in δ18O values in the basal Danian. Carbon isotopes show a negative excursion in δ13C values across the boundary. The shift toward more negative δ13C values begins 0.75 m (< 10 000 years) below the boundary clay at Kjolby Gaard. At Stevns Klint, lowest δ13C values correspond to an interval of low dinoflagellate abundance in the Danian. These δ13C patterns are attributed to a decrease in surface-water productivity, which began several thousand years before the K/T boundary event.

Genetic relations between the bivalves Cardium (Cerastoderma) edule, Cardium lamarcki and Cardium glaucum, studied by means of crossed immunoelectrophoresis

Rabbit antisera were raised against Cardium edule (L.) from the Waddensea locality, Skallingen, against Cardium lamarcki (Reeve) from the Danish locality, Dybso Fjord, and against Cardium glaucum (Bruguiere) from the French Mediterranean coast at Ile des Embiez. Allopatric and sympatric populations of C. lamarcki and C. edule from Swedish and Danish localities, and allopatric populations of C. glaucum from the Mediterranean Sea, Red Sea and Black Sea were studied by means of the described method and by starch gel electrophoresis of malate dehydrogenase (MDH). By means of crossed immuno-electrophoresis, using intermediary gels containing antibodies against one of the Cardium types and receiver gels containing antibodies against one of the others, type-specific and type-non-specific reactions were recorded. The results are compared with a similar investigation of Mytilus edulis, M. galloprovincialis and M. californianus. Based on immunological properties, it is shown that C. edule is a valid species different from the C. lamarcki/C. glaucum complex. It is further suggested that C. lamarcki and C. glaucum should still be referred to by these two names as significant genetic differences between them is revealed by this immunological study.

Dinoflagellate and Planktonic Foraminiferal Paleobathymetrical Indices in the Boreal Uppermost Cretaceous

A study has been carried out on the distribution of dinoflagellates and planktonic foraminifera in three Upper Maastrichtian borehole cores from Sweden and two uppermost Maastrichtian outcrop sections from Denmark. Several dinoflagellate variables display time-related changes in the Swedish cores. Likewise, the assemblage of planktonic foraminifers changes upsection. These trends may reflect the lowering of sea level that has been documented in the Upper Maastrichtian; the data suggest a change from a lower shelf to an upper shelf environment. Comparisons of these variables between the Swedish and Danish localities point to deeper depositional conditions in Denmark than in southern Sweden. The results indicate that the abundance of Spiniferites (both absolute and relative), the dinoflagellate diversity, and the G. multispina/H. striata ratio, are useful paleobathymetrical indices in the uppermost Cretaceous of boreal areas.

Dinoflagellate stratigraphy of the uppermost Danien to Ypresian in the Viborg 1 borehole, central Jylland, Denmark

Dinoflagellate cysts in the uppermost Danian to Lower Eocene section in the cored Viborg 1 borehole are described. The stratigraphical distribution of the dinoflagellates is shown; eight informal dinoflagellate zones are suggested. The dinoflagellate assemblages in sediments from various Danish localities are correlated with the Viborg 1 cores. A correlation with assemblages from other European Paleocene-Lower Eocene deposits is proposed. Two new species are described, and two new combinations are proposed.

Experimental field studies on the ecology of Scytosiphon lomentaria (Fucophyceae, Scytosiphonales) in Denmark

Experimental field studies on the ecology of Scytosiphon lomentaria have been carried out in two types of Danish localities during 1981 and 1982: a sheltered and shallow fiord with a scanty vegetation, and a more exposed locality with a rather dense vegetation. Colonization experiments have been made by depositing artificial substrates several times during spring and autumn in both localities. These showed that Scytosiphon lomentaria is able to colonize and develop, i.e. produce prostrate systems with erect thalli under long–day as well as under short–day conditions, and even to become fertile. Transplantation experiments between the two localities have been made in both directions in spring and autumn. These proved that the transplanted prostrate systems are able to sprout into erect thalli in accordance with the normal seasonal distribution in the new locality. The difference in seasonal occurrence is thus environmentally determined.Quantitative culture studies have confirmed that erect thalli develop independently of daylength.