Phylogenetic relationships of Echinostoma Rudolphi, 1809 (Digenea: Echinostomatidae) and related genera re-assessed via DNA and morphological analyses

In order to investigate the relationships within the Echinostomatidae two data sets of gene sequences were analysed. The first consisted of all previously published ND1 sequences (20) together with 17 new sequences. The latter represented six species from the cosmopolitan genera Echinostoma, Echinoparyphium, Hypoderaeum and Isthmiophora. The second data-set of ITS sequences again included all previously published sequences (12) and three new sequences from species of Echinostoma, Echinoparyphium and Isthmiophora. All new isolates, as well as voucher material from five previously sequenced isolates, were identified on the basis of morphological characters. The phylogenetic trees inferred from the ND1 data set helped to clarify the generic affiliation of all isolates and confirmed the morphological identifications. The only exception was Echinoparyphium aconiatum, whose current position in the genus Echinoparyphium was not supported by the sequence data. Although the ITS data provided insufficient resolution for an unequivocal solution to the relationships within the genus Echinostoma, it supported the identification of Echinoparyphium ellisi and the distinct species status of three isolates of Echinostoma revolutum as predicted from the ND1 data.     

The life-cycle of Echinostoma friedi n. sp. (Trematoda: Echinostomatidae) in Spain and a discussion on the relationships within the `revolutum' group based on cercarial chaetotaxy

The morphology of the different stages and life-cycle of Echinostoma friedi n. sp. are described and figured. The freshwater snail Lymnaea peregra (Gastropoda: Lymnaeidae) serves as the natural and experimental first intermediate host and L. corvus and Gyraulus chinensis (Gastropoda: Planorbidae) as experimental first intermediate hosts. These, and Physella acuta (Gastropoda: Physidae), also serve as second intermediate hosts. Adult worms, possessing 37 collar spines, were obtained from naturally infected Rattus norvegicus and experimentally from albino rats, golden hamsters and chickens. Mice were not suitable experimental definitive hosts. E. friedi differs from the most closely related species in the `revolutum' group mainly in terms of several morphological and biological features of the life-cycle stages and in its cercarial chaetotaxy. The chaetotaxy patterns of the species of the `revolutum' group are analyzed and the results show that a taxonomic comparison of these species may be carried out on the basis of the number of sensilla in the clusters CIII V₁, CIII V₂ (or CIII V₁ + CIII V₂), CIV DL and UV_b. These clusters appear adequate to establish taxonomic relationships between different species within the `revolutum' group.     

Production and chronobiology of emergence of the cercariae of Euparyphium albuferensis (Trematoda: Echinostomatidae)

The production and the chronobiology of emergence of the cercariae of Euparyphium albuferensis from Gyraulus chinensis experimentally infected with a single miracidium were established during 28 consecutive days from the first day of cercarial shedding. Moreover, the effect of a sudden change in light-dark cycling was investigated. Although the daily cercarial shedding rates show great variability, a progressive increase in cercarial production was observed in the first weeks of the cercarial shedding periods, probably in relation to the demography of intramolluscan larval stages. Under 12:12 light-dark cycling conditions, E. albuferensis cercariae emerged in the light, and the rhythm was circadian. The sudden change in the light-dark cycle resulted in corresponding alterations in emergence patterns, which shows that cercarial emergence was correlated to light-dark alternation. The ecological consequences of daily emergence by mobile cercariae whose target hosts are organisms that regularly occur in the same habitat with the molluscan emitting host, such as those of E. albuferensis, are discussed with reference to the hypotheses proposed to date, and an alternative hypothesis is proposed.     

Characterization of bacterial communities from activated sludge: Culture-dependent numerical identification versus in situ identification using group- and genus-specific rRNA-targeted oligonucleotide probes

The structures of bacterial communities were studied in activated sludge samples obtained from the aerobic and anaerobic zones of a wastewater treatment plant showing enhanced phosphorous removal. Samples were analyzed by in situ hybridization with oligonucleotide probes complementary to selected regions of the 16S and 23S ribosomal RNA (rRNA) characteristic for defined phylogenetic entities (genera and larger groups). The microbial community structures revealed by molecular techniques were compared with the compositions of culturable bacterial communities, obtained from the characterization of 255 isolates from tryptone-soy (TS) agar and R2A agar. These isolates were characterized by 89 physiological tests and their cellular fatty acid patterns, and identified. Culture-dependent techniques indicated the following distribution: different Aeromonas spp. (2.7–8.3% on R2A agar; 45.0–63.7% on TS agar), Acinetobacter spp. (5.4–9.0% on R2A agar; 5.0–9.1% on TS agar), Pseudomonas spp. (up to 10% on R2A agar) and Shewanella putrefaciens (up to 3.0% on R2A agar), all members of the gamma subclass of Proteobacteria, were isolated most frequently. The relatively rare isolates of the beta subclass were identified as Acidovorax spp., Alcaligenes spp., and Comamonas spp., The Gram-positive bacteria (high DNA G+C) were assigned mainly to Arthrobacter spp., Microbacterium spp., and Mycobacterium phlei. In order to assess the in situ abundance of the most frequently isolated genus, Aeromonas, two rRNA-targeted oligonucleotide probes were developed. The two gamma proteobacterial genera Aeromonas and Acinetobacter constituted less than 5% of all bacteria. In situ, Proteobacteria belonging to the beta subclass and high G+C Gram-positive bacteria were dominant. From filamentous bacteria, Sphaerotilus spp. and Leptothrix spp. could be detected occasionally. In addition, one sample contained a high proportion of the morphologically distinct filaments of Microthrix parvicella.As for the genus Acinetobacter, the relative abundance of the most frequently gamma-proteobacterial genus Aeromonas was overestimated by the intrinsic selectivity of cultivation. Cultivation on nutrient-rich medium (TS-agar) especially supported an enhanced isolation of bacteria belonging to these two genera. Correspondence to: P. Kämpfer.     

Chaetotaxy of members of the Gyrodactylidae (Monogenea), with comments upon their systematic relationships with the Monopisthocotylea and Polyopisthocotylea

This study investigates intra- and inter-generic variation in the chaetotaxy pattern of members of the Gyrodactylidae and compares these data with those from other monogenean groups elucidated by other workers. The staining of surface sensillary structures has revealed areas of homology between the two genera Gyrodactylus and Ooegyrodactylus, which have markedly contrasting modes of reproduction. Although species specific maps have been elucidated for members of the genus Gyrodactylus, no evolutionary trends in their sensillary patterns could be determined. Members of the family Gyrodactylidae are shown to possess a cluster of 3 × 2 sensilla, similar in their size and arrangement to the post-ocular (spo) cluster of sensilla in polyopisthocotyleans, although on the opposite surface. A comparable cluster of sensilla is lacking in monopisthocotyleans. This information gathered from chaetotaxy supports the proposal that the order Gyrodactylidea is close to the polyopisthocotylean line of monogenean evolution.     

Microsporidian Parasites of Trematode Larvae from Aquatic Snails in West Malaysia

In a survey of microsporidian parasites of trematode larvae of Malaysian fresh-water snails, species of Nose-matidae were found 7 times in rediae from 425 Lymnaea rubiginosa, 6 times in rediae from 97 Indoplanorbis exustus and not at all in other snails. There were 3 species of Microsporida in all: one, found in Echinoparyphium dunni, Echinostoma hystricosum and Echinostoma malayanum was identified as Nosema eurytremae Canning; another, which was found in E. dunni and Echinostoma audyi, had paired nuclei (diplokaryon form) in sporogony only and was named a new species, Nosema vasicola sp.n.; the 3rd, from E. hystricosum, lacked diplokaryon nuclei throughout development, was considered to belong to a new genus and was named Unikaryon piriformis gen.n., sp.n. Nosema eurytremae was transmitted experimentally to Fasciola hepatica rediae in Lymnaea truncatula.     

Phylogenetic relationships of Heterovesicula cowani, a microsporidian pathogen of Mormon crickets, Anabrus simplex (Orthoptera: Tettigoniidae), based on SSU rDNA-sequence analyses

The microsporidium Heterovesicula cowani, discovered in 1985, was initially identified as Vairimorpha sp. because it produces two types of spores: Nosema-like diplokaryotic spores and Thelohania-like mononuclear meiospores. However, light and electron microscopy studies revealed characters that did not fit any known microsporidian genera, and a new monotypic genus Heterovesicula was erected. The goal of this study was to test the validity of the genus Heterovesicula by molecular characterization of H. cowani and to assess its phylogenetic relationships to other microsporidia from insects. DNA from spores stored at −32 °C since 1992 was isolated and PCR-amplified with V1-1492 primers to obtain a partial small subunit ribosomal RNA gene sequence of 1165 bp, which was submitted to GenBank (Accession No. EU275200). Neighbor joining, maximum parsimony and maximum likelihood analyses performed against 18 microsporidia sequences, placed H. cowani as a sister taxon to the Nosema–Vairimorpha clade. The consensus of these analyses suggests that the Heterovesicula–Nosema–Vairimorpha group forms a dichotomy with the Encephalitozoon spp. branch. Other microsporidia parasitizing Orthoptera fell into two unrelated (or distantly related) lineages of terrestrial microsporidia: the Liebermannia spp. branch forms a dichotomy with Orthosomella operophterae within the Endoreticulatus–Orthosomella–Liebermannia group; and the Paranosema spp. branch clusters together with the Tubulinosema–Systenostrema lineage. The minimum pairwise distance in Kimura-2-Parameter analysis among 18 analyzed sequences was 0.37, which supports well the generic status for Heterovesicula. The obtained phylogenetic trees suggest that H. cowani is related to the Vairimorpha necatrix group, but not to other insect microsporidia producing octospores.     

Schistosoma bovis: Variability of cercarial production as related to the snail hosts: Bulinus truncatus, B. wrighti and Planorbarius metidjensis

Development of Schistosoma bovis from Spain in different species or genus of intermediate hosts (Bulinus truncatus, B. wrighti and Planorbarius metidjensis) modifies cercarial productivity and its dynamics. From B. truncatus to B. wrighti and to P. metidjensis, cercarial productivity decreases while the length of the production period is increased. Variations in the dynamics are less obvious between the two species of Bulinus than between Bulinus spp. and P. metidjensis. In the latter the emission pattern is characterized by a 45–48-day production rhythm. These differences are explained in terms of larval demographic strategies and biotic capacities of the hosts. The validity of employing cercarial production as an indicator of host-parasite compatibility is discussed.     

Intra- and Interspecific Comparisons of Bacterial Diversity and Community Structure Support Coevolution of Gut Microbiota and Termite Host

We investigated the bacterial gut microbiota from 32 colonies of wood-feeding termites, comprising four Microcerotermes species (Termitidae) and four Reticulitermes species (Rhinotermitidae), using terminal restriction fragment length polymorphism analysis and clonal analysis of 16S rRNA. The obtained molecular community profiles were compared statistically between individuals, colonies, locations, and species of termites. Both analyses revealed that the bacterial community structure was remarkably similar within each termite genus, with small but significant differences between sampling sites and/or termite species. In contrast, considerable differences were found between the two termite genera. Only one bacterial phylotype (defined with 97% sequence identity) was shared between the two termite genera, while 18% and 50% of the phylotypes were shared between two congeneric species in the genera Microcerotermes and Reticulitermes, respectively. Nevertheless, a phylogenetic analysis of 228 phylotypes from Microcerotermes spp. and 367 phylotypes from Reticulitermes spp. with other termite gut clones available in public databases demonstrated the monophyly of many phylotypes from distantly related termites. The monophyletic “termite clusters” comprised of phylotypes from more than one termite species were distributed among 15 bacterial phyla, including the novel candidate phyla TG2 and TG3. These termite clusters accounted for 95% of the 960 clones analyzed in this study. Moreover, the clusters in 12 phyla comprised phylotypes from more than one termite (sub)family, accounting for 75% of the analyzed clones. Our results suggest that the majority of gut bacteria are not allochthonous but are specific symbionts that have coevolved with termites and that their community structure is basically consistent within a genus of termites.     

Tree diversity and distribution in undisturbed and human-impacted sites of tropical wet evergreen forest in southern Western Ghats, India

The composition, abundance, population structure and distribution patterns of all woody species ( 30 cm gbh) were investigated in an undisturbed and two adjacent human-impacted sites of a tropical wet evergreen forest in Kalakad National Park, Western Ghats, south India. Three 1-ha plots were established, one each in (i) an undisturbed site (named site UD), (ii) in a site selectively felled 35 years ago (site SF – small stems felled leaving the large trees (as shade) for developing it into a cardamom estate, on the failure of which the site was abandoned) and (iii) a frequently disturbed site (site FD – round woods logged for use in ovens for curing cardamom). These sites are 1 to 3 km apart in the same wet evergreen forest. In the three study plots a total of 2150 stems (mean density 716 ha^–1) covering 122 species in 89 genera and 41 families were enumerated. Species richness was greatest (85 species ha^–1) in the undisturbed site UD, intermediate (83) in SF and lowest (80) in FD. Tree density was greatest (855 stems ha^–1) in site SF, intermediate (720) in UD and lowest (575) in FD. The forest stand was exceptionally voluminous in site UD (basal area 94.64 m² ha^–1), intermediate (66.9 m²) in SF and least (61.7 m²) in FD, due to tree removal for fuel in the latter sites. Species composition and abundance patterns markedly varied between the three sites. In UD and SF, primary forest species (Cryptocarya bourdillonii , Cullenia exarillata Myristica dactyloides etc.) occurred in greater density. In FD heliophilic secondary forest species (Elaeocarpus venustus, Litsea wightiana, Viburnum punctatum and Vitex altissima) were abundant, while these were absent in UD and SF. The species–area curve did not reach an asymptote in any of the sites on the 1-ha scale. The stand population structure was clearly reverse J shaped in UD and SF, while small stems were 2- to 3-fold fewer in FD. Most trees exhibited clumped distribution of individuals on the 1-ha scale. Variation in the kind and richness of species and their abundance is related to human interference and the need for forest conservation is emphasized.     

Durettechina beveridgei n. g., n. sp. (Nematoda: Seuratidae) from Antechinus spp. (Dasyuridae: Marsupialia) from Australia

Durettechina beveridgei n. g., n. sp. (Nematoda: Seuratidae) is described from Antechinus flavipes (Dasyuridae) from Victoria and New South Wales. A single female from A. bellus from the Northern Territory may also be D. beveridgei. This new genus is compared with other genera of the Echinonematinae, to which it has been assigned. The genus has a unique body armature and most closely resembles Chabaudechina, in the armature of the cephalic bulb, but has four rather than five rows of hooks, and Linstowinema, in having body hooks on the cuticle of the anterior region, but has 18–22 hooks in each row rather than 14–16. The hooks of Durettechina are also smaller and have a less complex root morphology than those of Linstowinema. Durettechina resembles Seurechina and Chabaudechina in having caudal alae into which papillae extend, but differs from both these genera in the number and arrangement of the caudal papillae, as well as in the body armature. Durettechina, is most different from Bainechina, which has neither hooks on a cephalic bulb nor body hooks on the anterior region nor caudal alae.     

Survival and infectivity of Hypoderaeum conoideum and Euparyphium albuferensis cercariae under laboratory conditions.

The survival characteristics of the cercariae of Hypoderaeum conoideum and Euparyphium albuferensis (Trematoda: Echinostomatidae) at 20 degrees C and 30 degrees C are described, and the age dependency of their infectivity at 20 degrees C is studied to determine their respective transmission efficiencies. Cercarial survival was found to be age-dependent and was higher at 20 degrees C. For both cercariae, the maximum life-span was 26 h at 20 degrees C and 16 h at 30 degrees C, and their respective times to 50% mortality were similar at each temperature. Both cercariae seem to be well adapted to transmission in their natural habitat, though cercarial infectivity of H. conoideum was higher than that of E. albuferensis, this being correlated with their prevalences in nature. The age-dependency of cercarial survival may be related to steadily diminishing endogenous energy levels, though the delay in attaining maximum infectivity suggests that other factors not related to energy considerations are involved in the delimitation of the cercarial infective period. However, this latter observation may constitute an adaptative mechanism allowing cercarial dissemination.     

Delayed luminescence of Prorocentrum minimum under controlled conditions

Delayed luminescence (DL), also termed delayed fluorescence or delayed light emission, is the phenomenon of long-lived light emission by plants and cyanobacteria after being illuminated with light and put into darkness. Culture growth of three Prorocentrum minimum strains was studied with DL measurements. DL decay kinetics was measured from 1–60 s after a pulse of white light. The strains used were from the Adriatic Sea (PmK), from Chesapeake Bay, USA (D5), and from the Baltic Sea (BAL), cultured at salinity of 32, 16, and 8 (practical salinity scale), respectively. The strains differed in cell size and chlorophyll a content (PmK > D5 > BAL), as well as in DL parameters. The DL results were compared to standard measurements of culture density and carbon content (calculated from biovolumes). DL decay curves had a specific peak, which changed with culture growth and showed more similarities between the strains PmK and D5. The DL intensity increased with cell density and carbon content in a two-stage process, corresponding to the lag and exponential phases of growth. DL intensity was best correlated with carbon content irrespective of strain and is proposed as an estimate of biomass and for differentiating between lag and exponential phases of growth.     

A high-density genetic map of hexaploid wheat (Triticum aestivum L.) from the cross Chinese Spring × SQ1 and its use to compare QTLs for grain yield across a range of environments

A population of 96 doubled haploid lines (DHLs) was prepared from F₁ plants of the hexaploid wheat cross Chinese Spring × SQ1 (a high abscisic acid-expressing breeding line) and was mapped with 567 RFLP, AFLP, SSR, morphological and biochemical markers covering all 21 chromosomes, with a total map length of 3,522 cM. Although the map lengths for each genome were very similar, the D genome had only half the markers of the other two genomes. The map was used to identify quantitative trait loci (QTLs) for yield and yield components from a combination of 24 site × treatment × year combinations, including nutrient stress, drought stress and salt stress treatments. Although yield QTLs were widely distributed around the genome, 17 clusters of yield QTLs from five or more trials were identified: two on group 1 chromosomes, one each on group 2 and group 3, five on group 4, four on group 5, one on group 6 and three on group 7. The strongest yield QTL effects were on chromosomes 7AL and 7BL, due mainly to variation in grain numbers per ear. Three of the yield QTL clusters were largely site-specific, while four clusters were largely associated with one or other of the stress treatments. Three of the yield QTL clusters were coincident with the dwarfing gene Rht-B1 on 4BS and with the vernalisation genes Vrn-A1 on 5AL and Vrn-D1 on 5DL. Yields of each DHL were calculated for trial mean yields of 6 g plant^–1 and 2 g plant^–1 (equivalent to about 8 t ha^–1 and 2.5 t ha^–1, respectively), representing optimum and moderately stressed conditions. Analyses of these yield estimates using interval mapping confirmed the group-7 effects on yield and, at 2 g plant^–1, identified two additional major yield QTLs on chromosomes 1D and 5A. Many of the yield QTL clusters corresponded with QTLs already reported in wheat and, on the basis of comparative genetics, also in rice. The implications of these results for improving wheat yield stability are discussed.     

Relationships between occurrences of toxic Dinophysis species (Dinophyceae) and small phytoplanktons in Japanese coastal waters

Fluctuations of the genus Dinophysis, which contained several toxic species of diarrhetic shellfish poisoning (DSP), were investigated during blooms in Hiroshima Bay, Mutsu Bay and Ise Bay, Japan. The co-occurrences of small phytoplanktons (cryptophytes, other nanophytoplanktons, cyanobacteria and eukaryotic picophytoplanktons) were investigated to search for relationships with mixotrophic Dinophysis. Cryptophytes were divided into three size-groups based on length of their chloroplasts (>10, 5–10 and <5 μm) during counting by epifluorescence microscopy. Clear relationships were not found between the occurrences of Dinophysis spp. and nanophytoplanktons, cyanobacteria and eukaryotic picophytoplanktons. However, the fluctuations of small-sized cryptophytes (<5 μm) showed a close relationship with that of D. acuminata in Hiroshima Bay. In Mutsu Bay, small-sized cryptophytes also accompanied the first occurrence peak of Dinophysis spp. In Ise Bay, peaks of the occurrences of middle- and small-sized cryptophytes were observed 2–3 weeks before the peak of D. acuminata. These cryptophytes decreased rapidly with increase in D. acuminata. These results suggest the possibility that small-sized cryptophytes may be food organisms for mixotrophic Dinophysis, with the abundance of Dinophysis dependent on these cryptophytes.     

Evolution of Lycopodiaceae Inferred from Spacer Sequencing of Chloroplast rRNA Genes

Nucleotide sequences of a chloroplast rDNA region including 8 bp from the 3" end of 23S rDNA–ITS2–4.5S rDNA–ITS3–5S rDNA–ITS4 (approximately 800 bp) were determined in 25 species of Lycopodiaceae and two species of the genus Isoetes. The rate of molecular evolution of spacers significantly varied in different Lycopsida taxa. A phylogenetic analysis by the neighbor-joining (NJ) method revealed that the family Lycopodiaceae is monophyletic. The topology of phylogenetic trees suggests the isolation of four or probably five genera in family Lycopodiaceae. For these genera, synapomorphic indels were detected. The obtained data were compared with the results of phylogenetic analysis of Lycopsida with regard to other sequences. The relationships of taxa within the family Lycopodiaceae is discussed.     

Diversification of Lupinus (Leguminosae) in the western New World: derived evolution of perennial life history and colonization of montane habitats

Previous phylogenetic studies of Lupinus (Leguminosae) based on nuclear DNA have shown that the western New World taxa form a monophyletic group representing the majority of species in the genus, with evidence for high rates of recent diversification in South America following final uplift of the Andes 2–4 million years ago (Mya). For this study, three regions of rapidly evolving non-coding chloroplast DNA (trnL intron, trnS–trnG, and trnT–trnL) were examined to estimate the timing and rates of diversification in the western New World, and to infer ancestral states for geographic range, life history, and maximum elevation. The western New World species (5.0–9.3 Mya, 0.6–1.1 spp./My) comprise a basally branching assemblage of annual plants endemic to the lower elevations of western North America, from which two species-rich clades are recently derived: (i) the western North American perennials from the Rocky Mountains, Great Basin, and Pacific Slope (0.7–2.1 Mya, 2.0–5.9 spp./My) and (ii) the predominantly perennial species from the Andes Mountains of South America and highlands of Mexico (0.8–3.4 Mya, 1.4–5.7 spp./My). Bayesian posterior predictive tests for association between life history and maximum elevation demonstrate that perennials are positively correlated with higher elevations. These results are consistent with a series of one or more recent radiations in the western New World, and indicate that rapid diversification of Lupinus coincides with the derived evolution of perennial life history, colonization of montane habitats, and range expansion from North America to South America.     

Molecular and morphometric systematics of Diapterus (Perciformes,Gerreidae)

The fish belonging to Gerreidae are common representatives of the coastal ichthyofauna of the tropics and subtropics of the world. The genus Diapterus has a neotropical distribution and currently includes four species: two in the Atlantic Ocean [Irish mojarra D. auratus Ranzani 1842 and rhombic mojarra D. rhombeus (Cuvier 1829)] and two in the Pacific Ocean [short‐beaked mojarra D. brevirostris (Sauvage 1879) and golden mojarra D. aureolus (Jordan & Gilbert 1882)]. This genus has uncertain taxonomy based on the remarkable and dissimilar morphology of D. aureolus. We evaluated the taxonomy and phylogeny of Diapterus with statistical (canonical discriminant analysis) and phylogenetic analyses (maximum parsimony, maximum likelihood and Bayesian inference) of geometric morphometric data (relative warps) and molecular data (12Sr RNA, 16Sr RNA, COI and RAG1 sequences). Our results indicate that the genus Diapterus as we currently conceive it is a polyphyletic group: [(Gerres cinereus – Eucinostomus spp. (D. aureolus – (Eugerres spp. (D. auratus – (D. brevirostris – D. rhombeus)))))]. Therefore, a taxonomic rearrangement is needed. To preserve the monophyly of the group, our proposal is to remove D. aureolus and place it in a new monotypic genus. A neotype is designated for D. aureolus as well as an artificial identification key for the genera and the species treated in this article.     

Molecular characterisation of four echinostomes (Digenea: Echinostomatidae) from birds in New Zealand,with descriptions of <Emphasis Type="Italic">Echinostoma novaezealandense</Emphasis> n. sp. and <Emphasis Type="Italic">Echinoparyphium poulini</Emphasis> n. sp.

Morphological and molecular characterisation of echinostome specimens (Digenea: Echinostomatidae) recovered in one Anas platyrhynchos L. and one Cygnus atratus (Latham) (Anseriformes: Anatidae) from New Zealand revealed the presence of two known species, Echinostoma miyagawai Ishii, 1932 and Echinoparyphium ellisi (Johnston & Simpson, 1944) and two species new to science. Comparative morphological and phylogenetic analyses supported the distinct species status of Echinostoma novaezealandense n. sp. ex Branta canadensis (L.), A. platyrhynchos and C. atratus, and Echinoparyphium poulini n. sp. ex C. atratus. Echinostoma novaezealandense n. sp., a species of the “revolutum” species complex characterised by the possession of a head collar armed with 37 spines, keyed down to E. revolutum but was distinguished from the latter in having a much narrower body with almost parallel margins, longer oesophagus, wider cirrus-sac, larger seminal vesicle, much smaller ventral sucker, ovary, Mehlis’ gland and testes, more anteriorly located ovary and testes, and distinctly smaller eggs (81–87 × 42–53 vs 106–136 × 55–70 µm). This new species appears similar to Echinostoma acuticauda Nicoll, 1914 described in Australia but differs in having a longer forebody, more posteriorly located ovary and testes, and much smaller eggs (81–87 × 42–53 vs 112–126 × 63–75 µm). Echinoparyphium poulini n. sp. is differentiated from the four species of Echinoparyphium possessing 37 collar spines considered valid as follows: from E. chinensis Ku, Li & Chu, 1964 in having a much smaller body, four (vs five) angle spines and simple seminal vesicle (vs bipartite); from E. schulzi Matevosyan, 1951 in having a less robust body at a comparable body length, much smaller ventral sucker, ovary and testes, and longer but narrower eggs (87–109 × 50–59 vs 70–85 × 60–84 µm); and from the two smaller forms, E. serratum Howell, 1968 and E. aconiatum Dietz, 1909, in a number of additional metrical features correlated with body size and especially in the possession of much larger collar spines. Partial fragments of the mitochondrial nad1 and 28S rRNA genes were amplified for representative isolates of the four species and analysed together with sequences for Echinostoma spp. and Echinoparyphium spp. available on GenBank. Phylogenetic analyses based on the mitochondrial nad1 gene revealed congruence between the molecular data and species identification/delineation based on morphology; this was corroborated by the 28S rDNA sequence data.     

Helminth parasites of wild foxes (Vulpes vulpes L.) in The Netherlands

To study the helminth fauna of wild foxes (Vulpes vulpes) in The Netherlands, material was collected from 139 foxes. The following parasites were found. Cestodes: Taenia spp. 53.3%, Hymenolepis spp. 1.5%; Trematodes: Alaria alata 10.9%, Cryptocotyle lingua 3.6%, Euparyphium melis 1.5%, Apophallus donicus 0.7%; Nematodes: Toxocara canis 73.7%, Uncinaria stenocephala 59.9%, Capillaria aerophila 46.8%, C. plica 23.5%, Molineus patens 5.1%, Crenosoma vulpis 4.5%, Strongyloides spp. 0.7%. The tapeworms Mesocestoides spp. and Echinococcus spp. were not seen. No relationship was observed between worm burden and sex, time of year or place of origin.