Contracaecum rudolphii B: gene content, arrangement and composition of its complete mitochondrial genome compared with Anisakis simplex s.l

In the present study, we sequenced the complete mt genome (14,022 bp) of parasitic nematode Contracaecum rudolphii B and its structure and organization compared with Anisakis simplex s.l. The mt genome of C. rudolphii B is slightly longer than that of A. simplex s.l. (13,916 bp). C. rudolphii B mt genome is circular, and consists of 36 genes, including 12 genes for proteins, 2 genes for rRNA and 22 genes for tRNA. This genome contains a high A+T (70.5%) content. The mt gene order for C. rudolphii B is the same as those for A. simplex s.l., but it is distinctly different from other nematodes compared. The start codons inferred in the mt genome of C. rudolphii B are TTG and ATT. Six protein-coding genes use TAA as a stop codon whereas five genes use T and one genes use TAG as a termination codon. This pattern of codon usage reflects the strong bias for A and T in the mt genome of C. rudolphii B. Phylogenetic analyses using concatenated amino acid sequences of the 12 protein-coding genes, with three different computational algorithms (Bayes, ML and MP), all revealed distinct groups with high statistical support, indicating that C. rudolphii B and A. simplex s.l. is distinct but closely related species. These data provide additional novel mtDNA markers for studying the molecular epidemiology and population genetics of the C. rudolphii B, and should have implications for the molecular diagnosis, prevention and control of anisakidosis in humans and animals.     

Molecular identification of two strains of third-stage larvae of Contracaecum rudolphii sensu lato (Nematoda: Anisakidae) from fish in Poland

Contracaecum sp. larvae (L3) from fish were identified using nucleotide sequences of the internal transcribed spacers ITS-1 and ITS-2 of the ribosomal DNA. The nematode larvae originated from fish in a freshwater situation (crucian carp Carassius carassius, from Selment Wielki Lake in Mazury, northeastern Poland) and a brackish-water region (Caspian round goby Neogobius melanostomus from the Baltic Sea, Gdafisk Bay at the Polish coast). Two strains (Contracaecum rudolphii A and B) of Contracaecum rudolphii senso lato, a parasite common at the adult stage in fish-eating birds, were identified. In fish from the freshwater site, only the strain temporarily designated C. rudolphii B was identified; in the brackish-water region, both strains were found, suggesting that fish serve as paratenic host for both genotypes. Contracaecum rudolphii sensu lato has been recorded in several species of fish-eating birds in Poland, particularly in the great cormorant, Phalacrocorax carbo, in which the abundance is highest. The results, although based on a restricted number of larvae, suggest that the life cycles of both genotypes can be completed in the Polish region and that at least one of them, C. rudolphii B, can develop both in fresh and brackish water.     

Population biology of Contracaecum rudolphii sensu lato (Nematoda) in the great cormorant (Phalacrocorax carbo) from northeastern Poland

Parameters related to the occurrence, aggregation, and population structure of the nematode Contracaecum rudolphii Hartwich, 1964 , in the great cormorant ( Phalacrocorax carbo ) from northeastern Poland were analyzed. A total of 491 birds of different ages (adults, immatures, and nestlings) was examined; the cormorants studied were taken from both fresh- and brackish water habitats. Contracaecum rudolphii were found in stomachs of 454 birds (92.5%); the 46,244 nematodes included third- (L3) and fourth-stage larvae (L4), and sub-adult and adult females and males. The distribution of nematodes in the host population were highly aggregated. The occurrence of C. rudolphii was significantly dependent on the host's age and habitat, as well as on season; the proportion of development stages differed significantly depending on birds' age and season (the latter only in adult birds). The infrapopulations of C. rudolphii in the adult cormorants showed distinct density-dependent correlations: that is, as the infrapopulation size increased, the number of adult females C. rudolphii decreased, and the proportion of larvae and sub-adult females increased. A higher proportion of larvae and sub-adult females characterized the component population structure of the nematodes in the cormorant nestlings, compared with adult birds, probably because of immune system deficiency in the immature birds, coupled with the development of the nematode population. Seasonal changes in the C. rudolphii population, observed in the adult cormorants, were not related to seasonality of the L3 occurrence in food; instead, the changes are believed to have resulted from independent processes of elimination of the oldest nematodes and their replacement by larval stages that subsequently matured.     

<Emphasis Type="Italic">Contracaecum bioccai</Emphasis> n. sp. from the brown pelican <Emphasis Type="Italic">Pelecanus occidentalis</Emphasis> (L.) in Colombia (Nematoda: Anisakidae): morphology,molecular evidence and its genetic relationship with congeners from fish-eating birds

Contracaecum bioccai n. sp. is described from the brown pelican Pelecanus occidentalis (L.) in northern Colombia (Totumo Marsh) based on 20 enzyme loci studied using multilocus allozyme electrophoresis. Moreover, genetic relationships between the new taxon and related congeners are presented based on allozyme data-sets and sequence analyses (519 bp) of the mtDNA-cox2 gene. Fixed allele differences were found at some of the allozyme loci analysed in comparison with other Contracaecum spp. from pelicans and cormorants [i.e. the sibling species of the C. rudolphii Hartwich, 1964 complex, C. septentrionale Kreis, 1955, C. micropapillatum (Stossich, 1890), C. microcephalum (Rudolphi, 1809) and C. pelagicum Johnston & Mawson, 1942]. The genetic distance, at the allozyme level, between C. bioccai n. sp. and its congeners ranged from D ( Nei ) = 0.80 versus C. septentrionale to D ( Nei ) = 1.40 versus C. micropapillatum. The genetic distance at the mtDNA cox-2 level ranged, on average, from K-2P = 0.12 versus the C. rudolphii species complex to K-2P = 0.15 versus C. micropapillatum. An overall concordant tree topology, obtained from UPGMA and NJ tree analyses inferred from allozyme data, as well as from MP, UPGMA and NJ inferred from mtDNA-cox2 sequence analysis, showed C. bioccai n. sp. as a separated lineage to the other Contracaecum spp. A concordant result was also obtained by PCA analysis based on both the allozyme and mtDNA cox-2 data-sets. All of the tree topologies, derived from the phylogenetic analysis inferred from both allozymes and mtDNA data-sets, were in substantial agreement and depicted C. bioccai as closely related to the sibling species of the C. rudolphii complex (C. rudolphii A and C. rudolphii B) and C. septentrionale. Morphological analysis and a differential diagnosis based on male specimens of C. bioccai, which had been genetically characterised by both allozyme markers and mtDNA sequences analysis with respect to morphologically related congeners, enabled the detection of differences in a numbers of characters, including spicule length, the morphology of the distal end of the spicule and the distribution patterns of the distal caudal papillae.     

Molecular phylogenetics and diagnosis of Anisakis, Pseudoterranova, and Contracaecum from northern Pacific marine mammals

Individual specimens of Anisakis, Pseudoterranova, and Contracaecum collected from marine mammals inhabiting northern Pacific waters were used for comparative diagnostic and molecular phylogenetic analyses. Forty-eight new sequences were obtained for this study of 14 Anisakis taxa, 8 Pseudoterranova taxa, 4 Contracaecum taxa, and 4 outgroup species. Partial 28S (LSU) and complete internal transcribed spacer (ITS-1, 5.8S, ITS-2) ribosomal DNA was amplified by the polymerase chain reaction and sequenced. Sequences of ITS indicated that Pseudoterranova specimens from Zalophus californianus (California sea lion), Mirounga angustirostris (northern elephant seal), Phoca vitulina (harbor seal), Enhydra lutris (sea otter), and Eumetopias jubatus (Steller's sea lion) exactly matched P. decipiens s. str., extending the host and geographic range of this species. Anisakis from northern Pacific marine mammals were most closely related to members of the A. simplex species complex. Comparison of Anisakis ITS sequences diagnosed the presence of A. simplex C in 2 M. angustirostris hosts, which is a new host record. Anisakis specimens from Phocoena phocoena (harbor porpoise), Lissodelphis borealis (Pacific rightwhale porpoise), and E. jubatus included 3 ITS sequences that did not match any known species. Contracaecum adults obtained from Z. californianus were most closely related to C. ogmorhini s.l. and C. rudolphii, but ITS sequences of these Contracaecum specimens did not match C. ogmorhini s. str. or C. margolisi. These novel Anisakis and Contracaecum ITS sequences may represent previously uncharacterized species. Phylogenetic analysis of LSU sequences revealed strong support for the monophyly of Anisakinae, Contracaecum plus Phocascaris, Pseudoterranova, and Anisakis. Phylogenetic trees inferred from ITS sequences yielded robustly supported relationships for Pseudoterranova and Anisakis species that are primarily consistent with previously published phenograms based on multilocus electrophoretic data.     

Infection by Contracaecum rudolphii (Nematoda: Anisakidae) in the Neotropic cormorant Phalacrocorax brasilianus,and fishes from the estuary of the Valdivia river,Chile

Seasonal parameters of infection by the nematode Contracaecum rudolphii in the Neotropic cormorant Phalacrocorax brasilianus (Phalacrocoracidae) and their relation with feeding and infection in fishes from River Valdivia, Chile, were determined. The prevalences of infection in birds were similar during seasons, whereas mean intensity and percentages of adult gravid females were higher in spring and summer, respectively. For fishes no seasonal differences were found in infection. Cormorant diet varied seasonally in relation to fish prey consumed and this can be related to seasonal differences in infection parameters of birds. The high infection in birds should be considered as a potential risk for salmoniculture.     

Further studies on Contracaecum spasskii Mozgovoi, 1950 and C. rudolphii Hartwich, 1964 (sensu lato) (Ascaridida: Anisakidae) from piscivorous birds in China

Contracaecum spasskii Mozgovoi, 1950, collected from the great crested grebe Podiceps cristatus (Linnaeus) (Podicipediformes: Podicipedidae), is redescribed using both light and, for the first time, scanning electron microscopy. Contracaecum spasskii differs from its congeners by having marked transverse cuticular annulations, the length of the oesophagus and spicules, the ratio between the intestinal caecum and the ventricular appendix, the number and arrangement of male caudal papillae, and especially by the particular morphology of the lips and interlabia. Some previously unreported morphological features of C. spasskii are also revealed and others corrected. Contracaecum rudolphii Hartwich, 1964 (sensu lato) is also redescribed based on the specimens collected from the great cormorant Phalacrocorax carbo sinensis (Blumenbach) (Pelecaniformes: Phalacrocoracidae) from China. Based on the geographical perspective, the present Chinese material may represent the species C. rudolphii B.     

Two new species of Contracaecum Railliet & Henry, 1912 (Nematoda: Anisakidae), C. fagerholmi n. sp. and C. rudolphii F from the brown pelican Pelecanus occidentalis in the northern Gulf of Mexico

DNA sequencing of the nuclear ribosomal DNA internal transcribed spacers (ITS) and mitochondrial rrnS and cox2 genes, and analysis of polymorphisms in restriction profiles in the ITS and rrnS, were used to characterise anisakid nematodes belonging to Contracaecum Railliet & Henry, 1912 infecting the brown pelican Pelecanus occidentalis (L.) in Galveston Bay, Texas and Sarasota Bay, Florida. Molecular data led to the detection of two new species: Contracaecum fagerholmi n. sp., which was also supported by clear morphological evidence, and Contracaecum rudolphii F, a new cryptic species within the Contracaecum rudolphii Hartwich, 1964 complex. Bayesian phylogenetic analysis demonstrated that C. fagerholmi and C. rudolphii F form two well-separated clusters, with C. fagerholmi being closely related to Contracaecum bioccai Mattiucci et al., 2008 and C. rudolphii F being included in the C. rudolphii complex. C. fagerholmi can be readily differentiated morphologically from all of its congeners, other than C. microcephalum (Rudolphii 1809) and the five currently recognised members of the C. rudolphii complex (C. rudolphii A, B, C, D and E). C. fagerholmi differs from C. microcephalum in the length of the spicules and the shape of the distal tip of the spicules, and from C. rudolphii (sensu lato) in the shape and size of the ventro-lateral and dorsal lips and by having interlabia which are not distally bifurcate. Further studies are needed to determine which morphological characteristics can be used to distinguish the cryptic species of the C. rudolphii complex in order to assign them with formal names. The recovery of a third species, C. bioccai, from the brown pelican confirms its occurrence in this host and extends its known geographical distribution.     

Gastrointestinal helminths of fish-eating birds from Chiloe Island, Chile

Of four species of fish-eating birds from Chiloé Island (Chile), three were infected with eight species of helminths. Tetrabothrius sp. was found in Larus scoresbii. Tetrabothrius cylindraceus, Profilicollis antarcticus, Anomotaenia dominicanus, Stephanoprora denticulata, Capillaria sp. and P. antarcticus were found in Larus dominicanus. Contracaecum rudolphii and Corynosoma sp. infected Phalacrocorax olivaceus. With the exception of S. denticulata, C. rudolphii and Capillaria sp., the above helminths are reported for the first time from Chile.     

Levels of infection of gastric nematodes in a flock of great cormorants (Phalacrocorax carbo) from Lake Biwa, Japan

A high prevalence (86.7%) of various species of nematodes was observed in the stomach of great cormorants living in Lake Biwa, Japan. There were varying numbers of adults belonging to two common genera, Eustrongylides Jagerskiold 1909 (Nematoda: Dioctophymatidae) and Contracaecum Railliet & Henry 1912 (Nematoda: Anisakidae). The first included common adenophorean nematodes comprising a single species, Eustrongylides tubifex and the second comprised ascaroid nematodes that contained four named species: Contracaecum rudolphii Hartwich, 1964, Contracaecum microcephalum Yamaguti, 1961, Contracaecum multipapillatum Drasche, 1882 and Contracaecum chubutensis Garbin, 2008. After the prevalence and intensity of the infection had been noted, both types of nematodes were frequently observed to penetrate the mucosa and intrude into the wall of the glandular stomach, where they caused gross haemorrhage and ulceration. The Eustrongylides sp. was predominantly found in a nodular lesion of the proventricular wall, while Contracaecum spp. were observed either free in the lumen of the proventriculus or, on occasion, deeply penetrating its wall. Of the Contracaecum spp., C. rudolphii was the most prevalent. Grossly, large numbers of nematodes were present in infected stomachs (for C. rudolphii intensity was 1-34 and 3-57 nematodes in male birds and 1-21 and 1-32 in females; for C. microcephalum 1-2 and 1 in male birds and 1-2 in females; for C. multipapillatum 2 in male cormorants and no infection in females; for C. chubutensis 1-2 and 1 in male birds and 1-5 and 1 in females and for E. tubifex 1-5 nematodes in male birds and 2-8 in females). Ulcerative inflammation and hyperaemia were the most common pathological presentations, especially in areas that had been invaded by parasites. Microscopically, varying degrees of granulomatous inflammatory reactions were seen, in addition to degenerated nematodes which appeared to have deeply penetrated mucosal surfaces and were surrounded by fibrous connective tissues.     

Selected mitochondrial genes as species markers of the Arctic Contracaecum osculatum complex

This study, aimed at testing the hypothesis that some mitochondrial genes can serve as species-specific markers, involved a comparison of the sequence variance of selected mitochondrial DNA genes of the Arctic Contracaecum osculatum species (C. osculatum A, C. osculatum B and C. osculatum C). We compared differences between five complete (ND2, CYTB, ND3, ND4L and ND6) and three partial (CO1, CO3 and ND5) protein-coding genes. The total length of the sequence of each of the 13 specimens was 4830?bp. The sample consisted of C. osculatum L3 larvae collected from Reinhardtius hippoglossoides and Gadus ogac from the Barents Sea and Davis Strait. The K2P distance values between the species ranged within 0.06-0.12, the intraspecific variability (0.01-0.03) proving 3-6 times lower. The lowest interspecific divergence was observed between C. osculatum A and C. osculatum B, whereas the highest intraspecific diversity was typical of C. osculatum C. Among the C. osculatum species studied, the highest nucleotide diversity was recorded in the CYTB, CO3 and ND5 genes. These genes may be useful in species identification of the very closely related Contracaecum sibling species.     

Helminth fauna of the yellow-legged gull Larus cachinnans in Galicia, north-west Spain

Thirty-six helminth species were found in 324 gulls examined during June 1994 to February 1996 from different localities of Galicia: 25 trematodes (Brachylaima sp., Brachylecithum microtesticulatum, Cardiocephaloides longicollis, Cryptocotyle lingua, Cryptocotyle concavum, Diplostomum spathaceum, Echinostephilla virgula, Galactosomum phalacrocoracis, Gigantobilharzia acotylea, Gymnophallus deliciosus, Gynaecotyla longiintestinata, Himasthla elongata, Himasthla quissetensis, Knipowitschiatrema nicolai, Levinseniella (Levinseniella) propinqua, Maritrema gratiosum, Maritrema linguilla, Microphallus primas, Microphallus similis, Ornithobilharzia canaliculata, Parorchis acanthus, Phagicola minuta, Psilostomum brevicolle, Renicola sp. and Stephanoprora denticulata), four cestodes (Alcataenia micracantha, Microsomacanthus ductilis, Tetrabothrius (Oriana) erostris and Wardium cirrosa), six nematodes (Anisakis simplex, Contracaecum rudolphii, Cosmocephalus obvelatus), Eucoleus contortus, Paracuaria adunca and Tetrameres (Tetrameres) skrjabini) and one acanthocephalan (Arhythmorhynchus longicollis). Tetrabothrius erostris was the most prevalent species (79.6%), followed by C. obvelatus (47.8%), C. lingua (37.4%), G deliciosus (30.9%), G. longiintestinata (22.8%), P. adunca (21.9%), B. microtesticulatum (17.6%), E. contortus (14.5%) and M. similis (9.3%). Microphallus similis was the dominant species, with a Berger-Parker index (BP) of 0.32, followed by T. erostris (BP=0.10). All species presented an aggregated dispersion except G. acotylea and G. phalacrocoracis, which showed a random dispersion. Species that seem to have the greatest predilection for specific sites along the intestine are: C. longicollis and A. micracantha (first third), Brachylaima sp., M. similis and G. longiintestinata (last third) and A. longicollis (second half). Eight species are known to be pathogenic to commercially important fish or molluscan species and several are pathogenic to humans.     

Effects of ecohydrological variables on current and future ranges,local suitability patterns,and model accuracy in big sagebrush

Forecasting of species and ecosystem responses to novel conditions, including climate change, is one of the major challenges facing ecologists at the start of the 21st century. Climate change studies based on species distribution models (SDMs) have been criticized because they extend correlational relationships beyond the observed data. Here, we compared conventional climate‐based SDMs against ecohydrological SDMs that include information from process‐based simulations of water balance. We examined the current and future distribution of Artemisia tridentata (big sagebrush) representing sagebrush ecosystems, which are widespread in semiarid western North America. For each approach, we calculated ensemble models from nine SDM methods and tested accuracy of each SDM with a null distribution. Climatic conditions included current conditions for 1970–1999 and two IPCC projections B1 and A2 for 2070–2099. Ecohydrological conditions were assessed by simulating soil water balance with SOILWAT, a daily time‐step, multiple layer, mechanistic, soil water model. Under current conditions, both climatic and ecohydrological SDM approaches produced comparable sagebrush distributions. Overall, sagebrush distribution is forecasted to decrease, with larger decreases under the A2 than under the B1 scenario and strong decreases in the southern part of the range. Increases were forecasted in the northern parts and at higher elevations. Both SDM approaches produced accurate predictions. However, the ecohydrological SDM approach was slightly less accurate than climatic SDMs (?1% in AUC, ?4% in Kappa and TSS) and predicted a higher number of habitat patches than observed in the input data. Future predictions of ecohydrological SDMs included an increased number of habitat patches whereas climatic SDMs predicted a decrease. This difference is important for understanding landscape‐scale patterns of sagebrush ecosystems and management of sagebrush obligate species for future conditions. Several mechanisms can explain the diverging forecasts; however, we need better insights into the consequences of different datasets for SDMs and how these affect our understanding of future trajectories.     

Molecular evidence for the occurrence of Contracaecum rudolphii A (Nematoda: Anisakidae) in shag Phalacrocorax aristotelis (Linnaeus) (Aves: Phalacrocoracidae) from Sardinia (western Mediterranean Sea)

Specimens of Contracaecum rudolphii Hartwich, 1964 (Nematoda: Anisakidae) from Phalacrocorax aristotelis (Linnaeus) from the Archipelago of La Maddalena (Sardinia, western Mediterranean Sea) were characterised genetically and compared with C. rudolphii A sensu D’Amelio et al. 1990 and C. rudolphii B sensu D’Amelio et al. 1990 from Phalacrocorax carbo sinensis (Blumenbach) from north-eastern Italy, and with C. rudolphii C sensu D’Amelio et al. 2007 from Phalacrocorax auritus (Lesson) from west-central Florida, USA. The sequencing of the small subunit of the mitochondrial ribosomal RNA gene (rrnS) and by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the same gene and of the internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA) allowed the identification of all specimens of C. rudolphii from P. aristotelis as C. rudolphii A. The results confirmed that the definition of genetic markers, following the analysis of nuclear ribosomal and mitochondrial DNA, provides quick and practical diagnostic tools for the detection of the 3 sibling species of C. rudolphii. The occurrence of C. rudolphii in P. aristotelis is reported for the first time from the Mediterranean area, improving the picture of the dispersal patterns of the populations of these piscivorous birds, and confirming the existence of different and isolated populations between the North and South European waters.     

Anisakid infection in the European shag Phalacrocorax aristotelis aristotelis

A total of 100 specimens of the European shag Phalacrocorax aristotelis aristotelis were examined for anisakid infections in the south-east North Atlantic. The taxonomic status of the anisakid nematodes was studied by structural analysis using light and scanning electron microscopy. Two species, Contracaecum septentrionale and C. rudolphii A (Nematoda: Ascaridoidea), occurred in the European shag with high values of prevalence and mean intensities of infection. These constitute new host records for both parasites, widening their known geographical distribution. Although the histopathology associated with the infections indicated that the anisakids did not have the potential to cause bird death, the parasites are capable of contributing to and hastening avian mortality in the case of heavily-parasitized cormorants and when stressed by xenobiotics and/or through a synergetic effect by other bioagressors.     

Molecules, wing pattern and distribution: an approach to species delimitation in the "loxurina group" (Lepidoptera: Lycaenidae: Penaincisalia)

The wide range of morphological variations in the "loxurina group" makes taxa identification difficult, and despite several reviews, serious taxonomical confusion remains. We make use of DNA data in conjunction with morphological appearance and available information on species distribution to delimit the boundaries of the "loxurina" group species previously established based on morphology. A fragment of 635 base pairs within the mtDNA gene cytochrome oxidase I (COI) was analysed for seven species of the "loxurina group". Phylogenetic relationships among the included taxa were inferred using maximum parsimony and maximum likelihood methods. Penaincisalia sigsiga (Bálint et al), P. cillutincarae (Draudt), P. atymna (Hewitson) and P. loxurina (C. Felder & R. Felder) were easily delimited as the morphological, geographic and molecular data were congruent. Penaincisalia ludovica (Bálint & Wojtusiak) and P. loxurina astillero (Johnson) represent the same entity and constitute a sub-species of P. loxurina. However, incongruence among morphological, genetic, and geographic data is shown in P. chachapoya (Bálint & Wojtusiak) and P. tegulina (Bálint et al). Our results highlight that an integrative approach is needed to clarify the taxonomy of these neotropical taxa, but more genetic and geographical studies are still required.     

Bythotrephes longimanus in the Commonwealth of Independent States: variability, distribution and ecolog

Bythotrephes is presently extending its distribution to basins throughout Europe and North America. We used long term data (1880–1997) on Bythotrephes longimanus in Eurasian waterbodies to characterize its ecology and tolerance to different ecological factors. Depending on season and environmental conditions of the waterbody, Eurasian B. longimanus may undergo cyclomorphosis and can exist as two distinct forms ( B. longimanus or B. cederstroemi). Regional forms of the species ( B. l. var. balticus, B. l. var. it transcaucasicus, and B. l. var. arcticus) have also been described. Occurrence and density of Bythotrephes populations varies across geographical scales, and are apparently affected primarily by water temperature and salinity. Bythotrephes is limited to regions where water temperature ranges between 4 and 30 ^C, although the species prefers waters between 10 and 24 °C. Bythotrephes tolerates salinity values between 0.04 and 8.0¡, though it prefers water between 0.04 and 0.4¡. These factors strongly restrict Bythotrephes' distribution in warmer and more saline waters of the Commonwealth of Independent States (CIS), particularly in southern Russia and Kazakhstan. Local abundance and distribution of Bythotrephes is affected by abiotic factors, including basin morphometry and flushing rate, temperature, oxygen concentration and pH, as well as by lake trophic status and fish predation. Planktivorous fish may restrict habitat use by Bythotrephes to regions of basins where overlap between these groups is minimized. Rivers, canals, and ephemeral waterbodies serve as important modes of Bythotrephes' dispersal throughout the CIS and western Europe. Morphological adaptations (e.g. a long caudal process, cyclic parthenogenesis, and production of resting eggs) permit Bythotrephes to tolerate seasonally inhospitable conditions, assist in its dispersal and range extension, and minimize fish predation.     

Concentration and diversity of uncultured Legionella spp. in two unchlorinated drinking water supplies with different concentrations of natural organic matter

Two unchlorinated drinking water supplies were investigated to assess the potential of water treatment and distribution systems to support the growth of Legionella spp. The treatment plant for supply A distributed treated groundwater with a low concentration (<0.5 ppm of C) of natural organic matter (NOM), and the treatment plant for supply B distributed treated groundwater with a high NOM concentration (8 ppm of C). In both supplies, the water temperature ranged from about 10°C after treatment to 18°C during distribution. The concentrations of Legionella spp. in distributed water, analyzed with quantitative PCR (Q-PCR), averaged 2.9 (± 1.9) × 10(2) cells liter(-1) in supply A and 2.5 (± 1.6) × 10(3) cells liter(-1) in supply B. No Legionella was observed with the culture method. A total of 346 clones (96 operational taxonomical units [OTUs] with ≥97% sequence similarity) were retrieved from water and biofilms of supply A and 251 (43 OTUs) from supply B. The estimation of the average value of total species richness (Chao1) in supply A (153) was clearly higher than that for supply B (58). In each supply, about 77% of the sequences showed <97% similarity to described species. Sequences related to L. pneumophila were only incidentally observed. The Legionella populations of the two supplies are divided into two distinct clusters based on distances in the phylogenetic tree as fractions of the branch length. Thus, a large variety of mostly yet-undescribed Legionella spp. proliferates in unchlorinated water supplies at temperatures below 18°C. The lowest concentration and greatest diversity were observed in the supply with the low NOM concentration.