Morphological differences and molecular phylogenetic relationship of two tapeworm species,Moniezia expansa and Moniezia benedeni,collected from domestic ruminants in northern Vietnam

Moniezia expansa and M. benedeni are two common tapeworm species of domestic ruminants over the world. However, their morphological and molecular data are available for limited specimens from a few countries. In the present study, we compared morphological characteristics of these two species collected from goats and cattle in northern Vietnam and analyzed their phylogenetic relationship based on the 5.8S and second internal transcribed spacer (ITS2) of nuclear ribosomal DNA and the mitochondrial cytochrome c oxidase subunit I gene (cox1). The two species are clearly distinguishable from one another not only in the morphological appearance of the interproglottidal glands but also in the gross appearance of mature and gravid proglottids. Molecular analyses revealed that the 5.8S-ITS2 sequences of Vietnamese M. expansa were highly similar (99.7%) to the sequences from Japan and India, and made a common clade, which was clearly distinct from M. benedeni of Vietnam. For cox1 sequences, Vietnamese M. expansa showed a high similarity to and were grouped with the sequences from Ethiopia and some sequences from Senegal and China to make a common clade, which was separated from the remaining clades of Senegal and China. The cox1 sequences of M. benedeni from China, Vietnam, and Senegal were far distant (10.0–15.9%) from each other. The results of this study suggest that more sequence data of Moniezia species with details of morphological features from various geographical locations should be obtained to clarify the taxonomic status of Moniezia species.     

Molecular Evidence for Multiple Origins of the European Spined Loaches (Teleostei,Cobitidae)

We present a phylogenetic investigation of the Northern Clade, the major monophyletic clade within the freshwater fish family Cobitidae, one of the most prominent families of freshwater fishes found in Asian and European waters. Phylogenetic reconstructions based on the cytochrome b and RAG-1 genes show the genera Microcobitis, Sabanejewia, Koreocobitis and Kichulchoia as monophyletic groups. These reconstructions also show a Cobitis sensu lato and a Misgurnus sensu lato group. The Cobitis sensu lato group includes all species of Cobitis, Iksookimia, Niwaella and Kichulchoia, while the Misgurnus sensu lato group includes Misgurnus, Paramisgurnus and Koreocobitis. Although the monophyly of both the Cobitis sensu lato and Misgurnus sensu lato groups is supported, relationships within the groups are incongruent with current generic definitions. The absence of monophyly of most genera included in the Cobitis sensu lato group (Cobitis, Iksookimia and Niwaella) or their low genetic differentiation (Kichuchoia) supports their consideration as synonyms of Cobitis. Molecular phylogenies indicate that the Asian species of Misgurnus experienced a mitochondrial introgression from a lineage of Cobitis. We also find two nuclear haplotypes in the same Cobitis species from the Adriatic area that, in the absence of morphological differentiation, may indicate molecular introgression. Most lineages within the Northern Clade consist of species found in East Asia. However, some lineages also contain species from Europe and Asia Minor. The phylogenetic relationships presented here are consistent with previous studies suggesting an East Asian origin of the Northern Clade. According to the current distributions and phylogenetic relationships of the Misgurnus sensu lato and Cobitis clade lineages, particularly of M. fossilis and C. melanoleuca, the range expansion of East Asian species into Europe was most likely via Siberia into Northern and Central Europe. Phylogenetic analyses also show that the Cobitis sensu lato group consists of two clear subgroups (I and II), each presenting geographical differences. Subgroup I is distributed exclusively in East Asian drainages with an Eastern European offshoot (C. melanoleuca), whereas Subgroup II includes species widespread throughout Europe (including the Mediterranean), Asia Minor, the Black Sea and the Caucasus, with some lineages related to species restricted to East Asia.     

Flavonoids in the species of Cyrtomium (Dryopteridaceae) and related genera

Flavonoids in 19 Cyrtomium, three Cyrtogonellum and two Phanerophlebia taxa were surveyed. Major flavonoids were flavonol O-glycosides based on kaempferol, quercetin, and sometimes myricetin, and C-glycosylflavones, such as isovitexin, vitexin, isoorientin, orientin and their O-glycosides. The C-methylflavanones, farrerol and cyrtominetin, and their 7-O-glucosides were isolated from Cyrtomium devexiscapulae and Cyrtomium laetevirens. Flavanones have been reported from Cyrtomium falcatum sensu lato. Though C. falcatum sensu lato is divided into four taxa, i.e. C. falcatum subsp. falcatum, C. falcatum subsp. australe, C. falcatum subsp. littorale, and C. devexiscapulae, the occurrence of the flavanones was restricted to C. devexiscapulae, and they did not occur in C. falcatum sensu stricto.     

Phylogenetic reassessment of Hyaloscyphaceae sensu lato (Helotiales, Leotiomycetes) based on multigene analyses

Hyaloscyphaceae is the largest family in Helotiales, Leotiomycetes. It is mainly characterized by minute apothecia with well-differentiated hairs, but its taxonomic delimitation and infrafamilial classification remain ambiguous. This study performed molecular phylogenetic analyses using multiple genes including the ITS-5.8S rDNA, the D1–D2 region of large subunit of rDNA, RNA polymerase II subunit 2, and the mitochondrial small subunit. The primary objective was to evaluate the phylogenetic utility of morphological characters traditionally used in the taxonomy of Hyaloscyphaceae through reassessment of the monophyly of this family and its genera. The phylogenetic analyses inferred Hyaloscyphaceae as being a heterogeneous assemblage of a diverse group of fungi and not supported as monophyletic. Among the three tribes of Hyaloscyphaceae only Lachneae formed a monophyletic lineage. The presence of hairs is rejected as a synapomorphy, since morphologically diversified hairs have originated independently during the evolution of Helotiales. The true- and false-subiculum in Arachnopezizeae are hypothesized to have evolved through different evolutionary processes; the true-subiculum is likely the product of a single evolutionary origin, while the false-subiculum is hypothesized to have originated multiple times. Since Hyaloscyphaceae sensu lato was not resolved as monophyletic, Hyaloscyphaceae sensu stricto is redefined and only applied to the genus Hyaloscypha.     

Revision of the paraphyletic genus Koerneria Meyl, 1960 and resurrection of two other genera of Diplogastridae (Nematoda)

Recent inferences of phylogeny from molecular characters, as well as a reexamination of morphological and biological characters, reject the monophyly of the nematode genus Koerneria Meyl, 1960 (Diplogastridae). Here, Koerneria sensu lato is revised. The genus, which previously consisted of 40 species, is separated into three genera. Almost all of the transferred species are moved to the resurrected genus Allodiplogaster Paramonov & Sobolev in Skrjabin et al. (1954). Koerneria and Allodiplogaster are distinguished from each other by a weakly vs. clearly striated body surface, an undivided vs. divided stomatal cheilostom, and arrangement of the terminal ventral triplet of male genital papillae, namely in that v5 and v6 are paired and separated from v7 vs. v5–v7 being close to each other. Allodiplogaster is further divided into two groups of species, herein called the henrichae and striata groups, based on both morphological and life-history traits. The henrichae group is characterized by papilliform labial sensilla and male genital papillae, a conical tail in both males and females, and an association with terrestrial habitats and insects, whereas the striata group is characterized by setiform labial sensilla and male genital papillae, an elongated conical tail in both sexes, and an association with aquatic habitats. A second genus, Anchidiplogaster Paramonov, 1952, is resurrected to include a single species that is characterized by its miniscule stoma and teeth, unreflexed testis, and a distinct lack of male genital papillae or stomatal apodemes. Lastly, one further species that was previously included in Koerneria sensu lato is transferred to the genus Pristionchus Kreis, 1932. The revision of Koerneria sensu lato is necessitated by the great variability in its subordinate taxa, which occupy a variety of habitats, in addition to the increased attention to Diplogastridae as a model system for comparative mechanistic biology.     

<Emphasis Type="Italic">Taenia arctos</Emphasis> n. sp. (Cestoda: Cyclophyllidea: Taeniidae) from its definitive (brown bear <Emphasis Type="Italic">Ursus arctos</Emphasis> Linnaeus) and intermediate (moose/elk <Emphasis Type="Italic">Alces</Emphasis> spp.) hosts

Taenia arctos n. sp. (Cestoda: Cyclophyllidea: Taeniidae) is described from the brown bear Ursus arctos Linnaeus (definitive host) and moose/elk Alces spp. (intermediate hosts) from Finland (type-locality) and Alaska, USA. The independent status of the new species and the conspecificity of its adults and metacestodes have been recently confirmed by the mtDNA sequence data of Lavikainen et al. (2011; Parasitology International, 60, 289–295). Special reference is given to morphological differences between the new species and T. krabbei Moniez, 1879 (definitive hosts primarily canines for the latter), both of which use the moose/elk (Alces spp.) as intermediate hosts (the latter also uses Rangifer and perhaps other northern ruminants), and between the new species and T. ursina Linstow, 1893, both of which use the brown bear U. arctos as a definitive host. New morphological data are also provided for adults and cysticerci of T. krabbei. The analysis includes potentially useful morphometric features that have not been previously applied to Taenia spp.     

Identification of a New Borrelia Species among Small Mammals in Areas of Northern Spain Where Lyme Disease Is Endemic

The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several areas where Lyme disease is endemic in northern Spain. A low rate of infestation by Ixodes ricinus nymphs was found in the small mammal populations studied that correlated with the near-absence of B. burgdorferi sensu lato in 184 animals tested and with the lack of transmission of B. burgdorferi sensu lato to I. ricinus larvae that fed on them. In contrast, questing ticks collected at the same time and in the same areas were found to carry a highly variable B. burgdorferi sensu lato repertoire (B. burgdorferi sensu stricto, Borrelia garinii, Borrelia valaisiana, and Borrelia afzelii). Interestingly, the only isolate obtained from small mammals (R57, isolated from a bank vole) grouped by phylogenetic analyses with other Borrelia species but in a separate clade from the Lyme disease and relapsing fever organisms, suggesting that it is a new species. This new agent was widely distributed among small mammals, with infection rates of 8.5 to 12% by PCR. Moreover, a high seroprevalence to B. burgdorferi sensu lato was found in the animal sera, suggesting cross-reactivity between B. burgdorferi sensu lato and R57. Although small mammals do not seem to play an important role as reservoirs for B. burgdorferi sensu lato in the study area, they seem to be implicated in the maintenance of spirochetes similar to R57.     

Comparative morphology of Ostertagia mossi and Ostertagia dikmansi (trichostrongylidae) from Odocoileus virginianus and comments on other Ostertagia spp. from the Cervidae

The synlophes of Ostertagia mossi Dikmans, 1931 and O. dikmansi Becklund & Walker, 1968 were found to be identical supporting recognition of these as the major and minor morphotypes, respectively, of a putative species pair within the Ostertagiinae. The cervical synlophe included three narrowly spaced, continuous, parallel ridges laterally and three parallel but more widely spaced ridges in the dorsal and ventral fields (Type II and Type A patterns, respectively). The disposition of the synlophe in conjunction with a long oesophageo-intestinal valve, and attributes of the genital cone and spicules are characteristics shared among other Ostertagia spp. from cervids in the Holarctic region that allow recognition of a species group morphologically distinct from the Ostertagiinae of domestic ruminants. The morphological similarity of O. mossi/O. dikmansi, O. leptospicularis Assadov, 1953/O. kolchida Popova, 1937 and O. gruehneri Skrjabin, 1929/O. arctica Mitzkewitsch, 1929 is substantial, but these species can be differentiated by small but constant differences in the synlophe and genital complex of males; females cannot be easily distinguished. The morphological similarity of this cryptic complex of species and their relatively restricted distribution among cervids suggests a coevolutionary history for hosts and parasites in the Holarctic region.     

Phylogenetic taxonomy of Hymenochaete and related genera (Hymenochaetales)

Species delimitation in Hymenochaete sensu lato was evaluated based on partial nucLSU rDNA sequences, isoenzyme analyses and morphological data. Analyses of LSU data revealed Hymenochaete sensu stricto and Pseudochaete as distinct monophyletic genera. Transfer of Hydnochaete olivacea into genus Pseudochaete and Dichochaete setosa into genus Hymenochaete (as H. resupinata) was supported by the results of molecular analyses. A new species Hymenochaete koeljalgii from Tanzania was described. The species of genus Hymenochaete sensu stricto were divided into four well-supported clades possessing no distinctive morphological characters.     

Peroxide metabolism in the cestodes Hymenolepis diminuta and Moniezia expansa

The enzymes of hydrogen peroxide metabolism have been investigated in the cestodes H. diminuta and M. expansa. Neither catalase, lipoxygenase, glutathione peroxidase, NADH peroxidase nor NADPH peroxidase could be detected in homogenates of either species. However, both H. diminuta and M. expansa possessed a peroxidase which had a high affinity for reduced cytochrome c. The peroxidase was characterized by substrate and inhibitor studies and cell fractionation showed the enzyme to be located in the mitochondrial membrane fraction. The peroxidase could act as a substitute for catalase, by destroying metabolic hydrogen peroxide. Appreciable superoxide dismutase activity was found in M. expansa and H. diminuta and it is possible that this enzyme is the source of helminth hydrogen peroxide.     

Experimental studies on the cadmium accumulation in the cestode Moniezia expansa (Cestoda: Anoplocephalidae) and its final host (Ovis aries)

The tapeworm Moniezia expansa and naturally infected sheep were investigated with respect to their cadmium accumulation. Cadmium chloride (CdCl₂, 0.2 g) was added to 10 ml of distilled water and administered orally to the sheep every day for a period of 1 week. The cadmium content of M. expansa was lower than that in the liver tissues of sheep, although this difference was not significant. The highest mean cadmium concentrations were found in the liver of sheep infected with M. expansa (24.5 ± 11.5 mg kg⁻¹ dry weight). The mean cadmium concentration measured in M. expansa was 21.5 ± 19.2 mg kg⁻¹ dry weight, which was 31 and 1.5 times higher than levels determined in the muscle and kidney of the host, respectively, but 0.9 times lower than levels determined in the liver of host. Sheeps with M. expansa infection always had higher cadmium concentrations in the tissues (with the exception of the blood) than their uninfected conspecifics.     

Morphological studies on two marine colepid ciliates from Qingdao, China, Nolandia orientalis spec. nov. and Pinacocoleps similis (Kahl, 1933) comb. nov. (Ciliophora, Colepidae)

Recent studies have revealed that the “lower” marine ciliates are far more diverse than previously suspected. During a survey on the ciliate fauna in coastal waters of Qingdao, northern China, we have isolated about 30 new or poorly known taxa. In the present study two colepid species are investigated, Nolandia orientalis spec. nov. and Pinacocoleps similis (Kahl, 1933) comb. nov. (basionym: Coleps similis Kahl, 1933). Their morphology and infraciliature are documented based on living observations and silver impregnations. The new species Nolandia orientalis differs from the type species N. nolandi mainly in the structure of tier plates. The structure of the tier plates was also the basis for transferring Coleps similis Kahl, 1933 to the genus Pinacocoleps and for three further new combinations: Pinacocoleps heteracanthus (Noland, 1937) comb. nov. (basionym: Coleps heteracanthus Noland, 1937), P. spiralis (Noland, 1937) comb. nov. (basionym Coleps spiralis Noland, 1937) and Pinacocoleps arenarius (Bock, 1952) comb. nov. (basionym: Coleps arenarius Bock, 1952).     

A taxonomic revision of the Neoserica (sensu lato) pilosula group (Coleoptera,Scarabaeidae, Sericini)

Nine new species of the Neoserica (sensu lato) pilosula Moser, 1915, group are described from China: Neoserica curvipenis sp. n., N. emeishanensis sp. n., N. lincangensis sp. n., N. ludingensis sp. n., N. lushuiana sp. n., N. rangshuiensis sp. n., N. shennongjiaensis sp. n., N. tianeana sp. n., and N. weibaoshanica sp. n. The lectotype of Neoserica pilosula Moser, 1915, is designated. Habitus and male genitalia are illustrated, a key to the species of the group and a map of species distribution are given.     

Effects of leaf litter and its fungal colonization on the diet of Limnomysis benedeni (Crustacea: Mysida)

The strong invasive freshwater mysid Limnomysis benedeni, a detritivorous–herbivorous feeder, has a preference for small food particles, but also feeds on leaf litter. Here, we tested whether leaf litter consumption by L. benedeni depends on the tree species and leaf conditioning (two types of physical and biological leaf conditioning). At the physical leaf conditioning, L. benedeni was fed with shortly leached or extensively leached leaves of five tree species in laboratory food assays. The mysid consumed shortly leached leaves of Copper Beech, Lombardy Poplar, Common Oak, and especially White Willow, and did not feed on shortly leached Black Alder leaves. The consumption of extensively leached leaves by L. benedeni did not depend on the tree species. Overall, 74% of the variation of the leaf consumption by L. benedeni was explained by the significant interaction of the factors carbon content and polyphenol content of the leaves, caused the feeding strategy of L. benedeni. For the biological leaf conditioning, the mysids consumed to a high degree naturally conditioned leaves, followed by leaves colonized by one of three fungi, but oomycete-colonized leaf litter and autoclaved leaves were consumed at similar low levels. Our results indicate that L. benedeni feeds on different types of conditioned leaves to different extents, and therefore may affect leaf litter degradation in many invaded freshwaters.     

Molecular Identification and Analysis of Borrelia burgdorferi Sensu Lato in Lizards in the Southeastern United States

Lyme borreliosis (LB) group spirochetes, collectively known as Borrelia burgdorferi sensu lato, are distributed worldwide. Wild rodents are acknowledged as the most important reservoir hosts. Ixodes scapularis is the primary vector of B. burgdorferi sensu lato in the eastern United States, and in the southeastern United States, the larvae and nymphs mostly parasitize certain species of lizards. The primary aim of the present study was to determine whether wild lizards in the southeastern United States are naturally infected with Lyme borreliae. Blood samples obtained from lizards in Florida and South Carolina were tested for the presence of LB spirochetes primarily by using B. burgdorferi sensu lato-specific PCR assays that amplify portions of the flagellin (flaB), outer surface protein A (ospA), and 66-kDa protein (p66) genes. Attempts to isolate spirochetes from a small number of PCR-positive lizards failed. However, PCR amplification and sequence analysis of partial flaB, ospA, and p66 gene fragments confirmed numerous strains of B. burgdorferi sensu lato, including Borrelia andersonii, Borrelia bissettii, and B. burgdorferi sensu stricto, in blood from lizards from both states. B. burgdorferi sensu lato DNA was identified in 86 of 160 (54%) lizards representing nine species and six genera. The high infection prevalence and broad distribution of infection among different lizard species at different sites and at different times of the year suggest that LB spirochetes are established in lizards in the southeastern United States.     

New parasite identified in Blennius sanguinolentus Pallas, 1811: Schikhobalotrema longivesiculatum sp. n (Haplosplanchnidae Poche, 1925)]

Schikhobalotrema longivesiculatum sp.n from Blennius sanguinolentus Pallas, 1811 at Gulf of Gaeta (Italy) is described, figured and compared with other species of the genus Schikhobalotrema Skrjabin et Guschanskaja, 1955. Schikhobalotrema longivesiculatum differs from Schikhobalotrema obtusa (Linton, 1916) Skrjabin et Guschanskaja, 1955, especially for sucker ratio, lobed ovary, seminal vesicle coiled and in larger size of eggs; from Schikhobalotrema pomacentri (Manter, 1937) Skrjabin et Guschanskaja, 1955 for lobed ovary, elongated seminal receptacle, smaller vitelline follicles and coiled seminal vesicle; from Schikhobalotrema sparisomae (Manter, 1937) Skrjabin et Guschanskaja, 1955 for elongated seminal receptacle, coiled seminal vesicle and extent of escretory vesicle.     

Three new species of Lentinus from northern Thailand

There have been few studies on the taxonomy and biodiversity of the genus Lentinus in Thailand, which is a genus of edible mushrooms. Recently, collections from 17 sites in northern Thailand yielded 47 specimens of Lentinus sensu lato. Three were shown to be new species of Lentinus sensu stricto and Lentinus roseus, L. concentricus and L. megacystidiatus are introduced in this paper. The new species are described and illustrated with line drawings and are justified and compared with similar taxa. Furthermore, ITS sequence data do not match closely with any species presently lodged in GenBank.