Bank voles (Myodes glareolus) and house mice (Mus musculus musculus; M. m. domesticus) in Europe are each parasitized by their own distinct species of Aspiculuris (Nematoda, Oxyurida).

The molecular phylogeny and morphology of the oxyuroid nematode genus Aspiculuris from voles and house mice has been examined. Worms collected from Myodes glareolus in Poland, Eire and the UK are identified as Aspiculuris tianjinensis, previously known only from China, while worms from Mus musculus from a range of locations in Europe and from laboratory mice, all conformed to the description of Aspiculuris tetraptera. Worms from voles and house mice are not closely related and are not derived from each other, with A. tianjinensis being most closely related to Aspiculuris dinniki from snow voles and to an isolate from Microtus longicaudus in the Nearctic. Both A. tianjinensis and A. tetraptera appear to represent recent radiations within their host groups; in voles, this radiation cannot be more than 2 million years old, while in commensal house mice it is likely to be less than 10,000 years old. The potential of Aspiculuris spp. as markers of host evolution is highlighted.

The Status of Heligmosomoides americanus, Representative of an American Clade of Vole-Infecting Nematodes.

Heligmosomoides americanus is shown by molecular phylogenetic analysis of 3 nuclear (28S, ITS1, and ITS2) and 2 mitochondrial (cytochrome oxidase 1 and cytochrome b) loci to be a distinct species of heligmosomid nematode with a long-independent evolutionary history, and not a subspecies of Heligmosomoides polygyrus . Rather than being a recent arrival in North America, the species probably originated as a Beringian immigrant with the host vole Phenacomys, approximately 2 million years ago (MYA).

Identification of a new species of digenean Notocotylus malhamensis n. sp. (Digenea: Notocotylidae) from the bank vole (Myodes glareolus) and the field vole (Microtus agrestis).

Notocotylus malhamensis n. sp. is described from the caecum of the bank vole (Myodes glareolus) and the field vole (Microtus agrestis) from Malham Tarn Nature Reserve in North Yorkshire, UK. In total, 581 specimens were collected from rodents trapped at a wetland site (Tarn Fen) between July 2010 and October 2011 with a prevalence of 66·7% and mean intensity of 94·6 in the bank vole and 50% prevalence and a mean intensity of 4·3 in the field vole. This species appears to be most closely related to other previously described Notocotylus species infecting rodents in Europe but differs principally by the metraterm to cirrus sac ratio (1:1·5-1:1·2) in combination with a densely spinulated cirrus, simple caeca and a greater number of ventral glands in the lateral rows (14-17). The use of molecular differentiation was of limited use in this study due to a paucity of relevant information in the DNA sequence databases. However, the complete ITS1-5.8S rDNA-ITS2 and partial 28S gene sequences have been generated to provide a definitive tool for identification of this species in future studies. As far as we know this is the first report of a notocotylid infection in M. glareolus in the UK.

Differences in the ecology of Bartonella infections of Apodemus flavicollis and Myodes glareolus in a boreal forest.

The epidemiology of Bartonella species infecting Apodemus flavicollis and Myodes glareolus in a forest in Eastern Poland was followed for 2 years using mark-recapture. Infections could be acquired in any month, but prevalence, and probability of infection, peaked in the summer. There were significant differences in the pattern of infections between the two species. Both hosts were primarily infected as juveniles, but the probability of infection was highest for A. flavicollis, which, evidence suggests, experienced longer-lasting infections with a wider range of Bartonella genotypes. There was no evidence of increased host mortality associated with Bartonella, although the infection did affect the probability of recapture. Animals could become re-infected, generally by different Bartonella genotypes. Several longer lasting, poorly resolved infections of A. flavicollis involved more than 1 genotype, and may have resulted from sequential infections. Of 22 Bartonella gltA genotypes collected, only 2 (both B. grahamii) were shared between mice and voles; all others were specific either to A. flavicollis or to M. glareolus, and had their nearest relatives infecting Microtus species in neighbouring fields. This heterogeneity in the patterns of Bartonella infections in wild rodents emphasizes the need to consider variation between both, host species and Bartonella genotypes in ecological and epidemiological studies.

Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea).

Despite routine screening requirements for the notifiable fish pathogen Gyrodactylus salaris, no standard operating procedure exists for its rapid identification and discrimination from other species of Gyrodactylus. This study assessed screening and identification efficiencies under real-world conditions for the most commonly employed identification methodologies: visual, morphometric and molecular analyses. Obtained data were used to design a best-practice processing and decision-making protocol allowing rapid specimen throughput and maximal classification accuracy. True specimen identities were established using a consensus from all three identification methods, coupled with the use of host and location information. The most experienced salmonid gyrodactylid expert correctly identified 95.1% of G. salaris specimens. Statistical methods of classification identified 66.7% of the G. salaris, demonstrating the need for much wider training. Molecular techniques (internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP)/cytochrome c oxidase I (COI) sequencing) conducted in the diagnostic laboratory most experienced in the analysis of gyrodactylid material, identified 100% of the true G. salaris specimens. Taking into account causes of potential specimen loss, the probabilities of a specimen being accurately identified were 95%, 87% and 92% for visual, morphometric and molecular techniques, respectively, and the probabilities of correctly identifying a specimen of G. salaris by each method were 81%, 58% and 92%. Inter-analyst agreement for 189 gyrodactylids assessed by all three methods using Fleiss' Kappa suggested substantial agreement in identification between the methods. During routine surveillance periods when low numbers of specimens are analysed, we recommend that specimens be analysed using the ITS-RFLP approach followed by sequencing of specimens with a "G. salaris-like" (i.e. G. salaris, Gyrodactylus thymalli) banding pattern. During periods of suspected outbreaks, where a high volume of specimens is expected, we recommended that specimens be identified using visual identification, as the fastest processing method, to select "G. salaris-like" specimens, which are subsequently identified by molecular-based techniques.

A redescription of Protospirura muricola Gedoelst, 1916 (Nematoda: Spiruridae), a parasite of murid rodents.

The spirurid nematode Protospirura muricola Gedoelst, 1916 is redescribed from Acomys dimidiatus (Desmarest) from the St Katherine Protectorate, Sinai, Egypt. Egyptian material closely resembled specimens of P. muricola from African mammals re-examined in this study, as well as conforming to published reports of this species. P. muricola with two denticles on each lateral lobe of the pseudolabia and six pairs of postanal papillae is closest to P. pseudomuris Yokohata & Abe, 1989, but can be readily distinguished in having the right spicule shorter than the left. The significance of the characteristics of the head and mouth, and of the male spicules, in characterising Protospirura Seurat, 1914 is evaluated. P. muricola, an African parasite of rodents, appears to have spread globally with synanthropic rat final hosts and possibly with the cosmopolitan dermapteran intermediate host Leucophaea maderae (Fabr.).

Temporal and between-site variation in helminth communities of bank voles ( Myodes glareolus) from N.E. Poland. 1. Regional fauna and component community levels.

Helminth infections were studied in bank voles (Myodes glareolus) from 3 woodland sites in N.E. Poland in the late summers of 1999 and 2002, to assess the temporal stability of derived statistics describing the regional helminth fauna and component community structure, and spatial influence on the latter. Regional helminth fauna changed dramatically between the two years, primarily due to a fall in the abundance of Syphacia petrusewiczi but was partially compensated for by an increase in Mesocestoides lineatus and Cladotaenia globifera. It was dominated by nematodes overall, but more so in 1999 than in 2002 when larval cestodes were more frequent. Most derived parameters for component community structure varied considerably between sites and the two surveys, the hierarchical order for sites not being maintained between surveys. They were susceptible to the disproportionate influence of three relatively rare, unpredictable species with the greatest overall aggregated distribution among hosts. Jaccard's similarity index was less influenced by the rare species, showing greater stability between sites and across years. In conclusion, temporal variation confounded any site-specific characteristics of the summary measures quantified in this study and their usefulness is therefore restricted to the years in which the surveys were conducted.

Temporal and between-site variation in helminth communities of bank voles ( Myodes glareolus) from N.E. Poland. 2. The infracommunity level.

The relative importance of temporal and spatial effects was assessed in helminth communities of bank voles (Myodes glareolus) in 3 woodland sites in N.E. Poland in the late summers of 1999 and 2002. Among common species the rank order of sites in relation to prevalence and abundance of infection was maintained between surveys. Site effects accounted for most of the deviance (in statistical models), and time was less important, so the exact location from which voles were sampled was of critical importance. The only exception was Syphacia petrusewiczi. In contrast, for derived measures such as species richness and diversity, most deviance was accounted for by host age, and the interaction between site and year was significant, implying that rank order of sites changed between years. Temporal effects on derived measures were generated primarily by a combination of relatively small changes in prevalence and abundance of the common, rather than the rare, species between the years of the study. In the medium-term, therefore, helminth communities of bank voles in N.E. Poland had a stable core, suggesting a substantial strong element of predictability.

The uses of helium and xenon in current clinical practice.

The noble gases have always been an enigma. Discovered late in the history of chemistry and in seemingly small quantities in our atmosphere, they are some of the most unreactive elements known. However, despite being extremely inert, the noble gases (helium, neon, argon, krypton, xenon and radon) have found diverse and ever expanding applications in medicine. Of all of them, the gases that have found the greatest number of uses in the field of anaesthesia and related specialties are helium and xenon. This review focuses on the history of the discovery of both gases, their unique physicochemical properties and describes their uses in clinical practice with particular emphasis on those applicable to anaesthesia.

The biology of gyrodactylid monogeneans: the "Russian-doll killers".

This article reviews the history of gyrodactylid research focussing on the unique anatomy, behaviour, ecology and evolution of the viviparous forms while identifying gaps in our knowledge and directions for future research. We provide the first summary of research on the oviparous gyrodactylids from South American catfish, and highlight the plesiomorphic characters shared by gyrodactylids and other primitive monogeneans. Of these, the most important are the crawling, unciliated larva and the spike sensilla of the cephalic lobes. These characters allow gyrodactylids to transfer between hosts at any stage of the life cycle, without a specific transmission stage. We emphasise the importance of progenesis in shaping the evolution of the viviparous genera and discuss the relative extent of progenesis in the different genera. The validity of the familial classification is discussed and we conclude that the most significant division within the family is between the oviparous and the viviparous genera. The older divisions into Isancistrinae and Polyclithrinae should be allowed to lapse. We discuss approaches to the taxonomy of gyrodactylids, and we emphasise the importance of adequate morphological and molecular data in new descriptions. Host specificity patterns in gyrodactylids are discussed extensively and we note the importance of host shifts, revealed by molecular data, in the evolution of gyrodactylids. To date, the most closely related gyrodactylids have not been found on closely related hosts, demonstrating the importance of host shifts in their evolution. The most closely related species pair is that of G. salaris and G. thymalli, and we provide an account of the patterns of evolution taking place in different mitochondrial clades of this species complex. The host specificity of these clades is reviewed, demonstrating that, although each clade has its preferred host, there is a range of specificity to different salmonids, providing opportunities for complex patterns of survival and interbreeding in Scandinavia. At the same time, we identify trends in systematics and phylogeny relevant to the G. salaris epidemics on Atlantic salmon in Norway, which can be applied more generally to parasite epidemiology and evolution. Although much of gyrodactylid research in the last 30 years has been directed towards salmonid parasites, there is great potential in using other experimental systems, such as the gyrodactylids of poeciliids and sticklebacks. We also highlight the role of glacial lakes and modified river systems during the ice ages in gyrodactylid speciation, and suggest that salmon infecting clades of G. salaris first arose from G. thymalli in such lakes, but failed to spread fully across Scandinavia before further dispersal was ended by rising sea levels. This dispersal has been continued by human activity, leading to the appearance of G. salaris as a pathogen in Norway. We review the history and current status of the epidemic, and current strategies for elimination of the parasite from Norway. Finally, we consider opportunities for further spread of the parasite within and beyond Europe.

A case of mistaken identity--reappraisal of the species of canid and felid hookworms (Ancylostoma) present in Australia and India.

This study serves to clarify the current status of canid and felid Ancylostoma species present in Australia. The morphological identification of A. ceylanicum from cats for the first time in Townsville, Australia, appears to be in error, together with the genetic markers provided for the species. Morphological and genetic data presented herein provide strong evidence that the hookworms from cats in Towsville are not A. ceylanicum as previously identified (i.e. the first report of this species in Australia), but are A. braziliense. Therefore the subsequent genetic markers established for A. ceylanicum in subsequent molecular studies based on these Townsville specimens should also be attributed to A. braziliense. Based on this information, a study of canine hookworm species present in northern India is also in error and it is apparent that the hookworms found in this region are those of A. ceylanicum. The distribution of A. braziliense and A. ceylanicum in the Americas and Asia Pacific region is discussed together with the importance of combining parasite morphology with genetic data for parasite diagnosis in epidemiological studies.

Molecular evidence that Heligmosomoides polygyrus from laboratory mice and wood mice are separate species.

The gastro-intestinal (GI) nematode Heligmosomoides polygyrus is an important experimental model in laboratory mice and a well-studied parasite of wood mice in the field. Despite an extensive literature, the taxonomy of this parasite in different hosts is confused, and it is unclear whether laboratory and field systems represent the same or different Operational Taxonomic Units (OTUs). Molecular analyses reveal high sequence divergence between H. p. bakeri (laboratory) and H. p. polygyrus (field); 3% difference in the ribosomal DNA Internal Transcribed Spacers (ITS) and 8.6% variation in the more rapidly evolving mitochondrial cytochrome c oxidase I (COI) gene. The COI sequence of U.K. H. p. polygyrus is more similar to H. glareoli from voles than to H. p. bakeri, while a single isolate of H. p. polygyrus from Guernsey confirms the extent of genetic variation between H. p. polygyrus populations. Analysis of molecular variance demonstrated that mtCOI sequence variation is associated primarily with groups with distinct ITS2 sequences, and with host identity, but is not partitioned significantly with a single combined taxon H. polygyrus incorporating European and North American isolates. We conclude therefore that the laboratory OTU should be raised to the level of a distinct species, as H. bakeri from the laboratory mouse Mus musculus, and we reject the hypothesis that H. bakeri has diverged from H. polygyrus in the recent past following introduction into America. However, we are unable to reject the hypothesis that H. polygyrus and H. bakeri are sister taxa, and it may be that H. polygyrus is polyphyletic or paraphyletic.

Gyrodactylus pictae n. sp. (Monogenea: Gyrodactylidae) from the Trinidadian swamp guppy Poecilia picta Regan, with a discussion on species of Gyrodactylus von Nordmann, 1832 and their poeciliid hosts.

Gyrodactylus pictae n. sp. is recorded from Poecilia picta in heterospecific shoals with the guppy P. reticulata in Northern Trinidad. G. pictae is morphologically similar to G. turnbulli Harris, 1986, but the hamuli and marginal hooks are slightly smaller and more gracile. The toe and the point of the marginal hook have a distinctly different shape, providing the best morphological characters for distinguishing the two species. The rDNA ITS1 and ITS2 sequences differ from those of G. turnbulli (the closest relative) by >5, suggesting that these two taxa are not sibling species. The origin of the two species on poeciliids of the subgenus Micropoecilia is discussed, and it is suggested that this may represent a case of host-parasite co-evolution.

Variation in the helminth community structure in spiny mice (Acomys dimidiatus) from four montane wadis in the St Katherine region of the Sinai Peninsula in Egypt.

We compared helminth communities in spiny mice (Acomys dimidiatus) from 4 wadis in the arid montane region of the southern Sinai in Egypt, in a 4-week period in late summer. Total helminth species richness was 14 (8 nematodes, 5 cestodes and 1 acanthocephalan) with 94% of mice carrying at least 1 species and an overall mean species richness of 1.85. The most prevalent parasites were Protospirura muricola (47.8 %) and Dentostomella kuntzi (46.3%). One larval cestode, Joyeuxiella rossicum, represents a new host record. The helminth community was dominated by intestinal nematodes (88.7%) of which 58.2% were arthropod-transmitted heteroxenic species. At the component community level, 70% of the worms were recovered from mice in just two wadis (Gharaba and Tlah) and 48.6 % of intestinal nematodes were from Wadi Gharaba. Although only 7 species of helminths were recorded from Wadi Gharaba, this site gave the highest Berger-Parker dominance index because of P. muricola. P. muricola was also dominant in Wadi El Arbaein whilst Syphacia minuta was the dominant species in Wadis Gebal and Tlah. At the infracommunity level, mean species richness and Brillouin's index of diversity were highest in Wadi Tlah and lowest in Wadi Gebal, and the former was age dependent. Whilst mice from different wadis differed in the nematodes that were most common, those from Wadi Gharaba carried the highest mean number of worms/mouse. The abundance of P. muricola in particular varied markedly between sites: Wadi Gharaba was distinct as the site showing the highest mean worm burden whereas mice from Wadi Gebal were uninfected. None of the directly transmitted oxyuroid nematodes showed significant variation in abundance between wadis, or host sex or age classes. Overall, the single extrinsic factor in the study, site of capture, was more important than the intrinsic factors in explaining variation in helminth communities in the region. We conclude that in the high mountains of southern Sinai, each wadi is distinct in terms of its rodent parasites, and hence we expect spatially different coevolutionary pressures on their hosts, with resultant variation in life-histories.

Nominal species of the genus Gyrodactylus von Nordmann 1832 (Monogenea: Gyrodactylidae), with a list of principal host species.

The total diversity of the monogenean genus Gyrodactylus is evaluated. There are 409 potentially valid species names within the genus, recorded from c. 400 host species. Five species have been placed within Fundulotrema and an additional 51 Gyrodactylus species names represent synonyms, nomina nuda or have been reassigned to other non-viviparous monogenean genera. While the majority of Gyrodactylus species (59%) are recorded from single hosts, some have a much broader broad range.

Susceptibility of Baltic and East Atlantic salmon Salmo salar stocks to Gyrodactylus salaris (Monogenea).

The susceptibility of a Baltic salmon stock Salmo salar (Indalsälv, central Sweden) to Norwegian Gyrodactylus salaris (Figga strain, central Norway) was experimentally tested and compared with previously obtained results on East Atlantic salmon (Lierelva, SE Norway). Contrary to expectation, the Baltic salmon, which had no prior exposure to this parasite strain, appeared almost as susceptible as the Norwegian salmon parr that naturally experience G. salaris-induced mortality. Individually isolated salmon of both stocks sustained G. salaris infections with little evidence of innate resistance. A few individuals of the Indalsälv stock controlled their infection from the beginning, but overall there was considerable heterogeneity in the course of infection in both stocks. On individual hosts, G. salaris growth rates declined steadily throughout the infection, a trend which was particularly marked amongst the Lierelva stock. On shoaling Lierelva fish, there was some evidence of reduced parasite population growth towards the end of the infection; this was not apparent in Indalsälv fishes. These results reflect a growing awareness that not all Baltic salmon may be resistant to Norwegian G. salaris, and that Norwegian and Baltic G. salaris strains may differ in virulence. Consequently, management decisions concerning this parasite-host system should be based upon the actual, and tested, susceptibility of stocks under consideration and not upon identification of stocks as either Atlantic or Baltic.

Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds.

BACKGROUND: Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes. OBJECTIVES: To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components. METHODS: Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8-9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4-amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the 'chymotrypsin-like' serine proteinase. CONCLUSIONS: These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin-like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components.

Use of ITS rDNA for discrimination of European green- and brown-banded sporocysts within the genus Leucochloridium Carus, 1835 (Digenea: Leucochloriidae).

Transmission of Leucochloridium species to their definitive avian hosts may be facilitated by the rhythmic movement of coloured sporocyst broodsacs in the ocular tentacles of infected snails. These broodsacs resemble caterpillars and by enticement increase the probability of predation by birds. Broodsac banding pattern and colour (green, yellow to red/brown) have traditionally formed part of the taxonomic criteria for the genus. In this study, sequence divergence of the 5.8S rDNA gene and associated internal transcribed spacers (ITS1 and ITS2) in two of the most frequently observed Leucochloridium taxa from Europe is related to broodsac type based on colour and banding pattern. The present green-banded broodsac (L. paradoxum Carus, 1835) and brown-banded broodsac forms (L. variae McIntosh, 1932) differ in ITS sequence by 6.8%, confirming their distinctness. No intraspecific differences were noted within each colour morph in specimens collected from Poland, Denmark or Norway, indicating that a single taxon of each type occurs in Europe. The significance of these findings to our understanding of metapopulation dynamics and evolutionary ecology of Leucochloridium is discussed.