Comparative molecular analyses of Eimeria Schneider (Apicomplexa: Eimeriidae) species from rock ptarmigan in Iceland, Svalbard-Norway, and Japan.

The rock ptarmigan (Lagopus muta) has a Holarctic breeding distribution and is found in arctic and sub-arctic regions. Isolated populations and glacial relicts occur in alpine areas south of the main range, like the Pyrenees in Europe, the Pamir mountains in Central Asia, and the Japanese Alps. In recent decades considerable effort has been made to clarify parasite infections in the rock ptarmigan. Seven Eimeria spp. have been reported parasitizing rock ptarmigan. Two of those species, E. uekii and E. raichoi parasitizing rock ptarmigan (L. m. japonica) in Japan, have been identified genetically. Here we compare partial sequences of nuclear (18S rRNA) and mitochondrial (COI) genes and we detail the morphology of sporulated oocysts of E. uekii and E. raichoi from Japan, E. muta and E. rjupa, from the rock ptarmigan (L. m. islandorum) in Iceland, and two undescribed eimerian morphotypes, Eimeria sp. A, and Eimeria sp. B, from rock ptarmigan (L. m. hyperborea) in Norway (Svalbard in the Norwegian Archipelago). Two morphotypes, ellipsoidal and spheroidal, are recognized for each of the three host subspecies. Our phylogenetic analysis suggests that the ellipsoidal oocyst types, E. uekii, E. muta, and Eimeria sp. A (Svalbard-Norway) are identical and infects rock ptarmigan in Japan, Iceland, and Svalbard-Norway, respectively. Eimeria uekii was first described in Japan in 1981 so that E. muta, described in Iceland in 2007, and Eimeria sp. A in Svalbard-Norway are junior synonyms of E. uekii. Also, phylogenetic analysis shows that the spheroidal oocyst types, E. rjupa and Eimeria sp. B (Svalbard-Norway), are identical, indicating that rock ptarmigan in Iceland and Svalbard-Norway are infected by the same Eimeria species and differ from E. raichoi in Japan.

First species record of Strigea falconis Szidat, 1928 (Trematoda, Strigeidae) from gyrfalcon Falco rusticolus in Iceland-pros and cons of a complex life cycle.

Strigea falconis is a common parasite of birds of prey and owls widely distributed in the Holarctic. We aimed to characterise S. falconis from Iceland via integrative taxonomic approach and to contribute to the understanding of its circulation in the Holarctic. We recovered adult S. falconis from two gyrfalcons (Falco rusticolus) collected in 2011 and 2012 in Iceland (Reykjanes Peninsula, Westfjords) and characterised them by morphological and molecular genetic (D2 of rDNA, cox1, ND1 of the mDNA) methods. We provide the first species record of S. falconis in Iceland which to the best of our knowledge is its northernmost distributional range. The presence of S. falconis in Iceland is surprising, as there are no suitable intermediate hosts allowing completion of its life cycle. Gyrfalcons are fully sedentary in Iceland; thus, the only plausible explanation is that they acquired their infection by preying upon migratory birds arriving from Europe. Our data indicate that the most likely candidates are Anseriformes and Charadriiformes. Also, we corroborate the wide geographical distribution of S. falconis, as we found a high degree of similarity between our haplotypes and sequences of mesocercariae from frogs in France and of a metacercaria from Turdus naumanni in Japan, and adults from Buteo buteo and Circus aeruginosus from the Czech Republic. The case of Strigea falconis shows the advantages of a complex life cycle and also depicts its pitfalls when a parasite is introduced to a new area with no suitable intermediate hosts. In Iceland, gyrfalcons are apparently dead-end hosts for S. falconis.

Wood mouse (Apodemus sylvaticus L.) as intermediate host for Mesocestoides canislagopodis (Rudolphi, 1810) (Krabbe 1865) in Iceland.

Mesocestoides canislagopodis is a common parasite of the arctic fox (Vulpes lagopus) in Iceland. In the past, household dogs (Canis familiaris) and cats (Felis catus) were also reported in Iceland to be infected. Recently, scolices of a non-maturing Mesocestoides sp. were detected in the intestines of the gyrfalcon (Falco rusticolus), and tetrathyridia were isolated in the body cavity of rock ptarmigan (Lagopus muta) and subsequently described. All stages were confirmed, using both morphological and molecular methods, to belong to the same species, M. canislagopodis. In the present study, post-mortem examination of wood mice (Apodemus sylvaticus), sampled in autumn 2014 on a farm in Northeast Iceland, revealed the presence of tetrathyridia in the peritoneal cavity and in the liver. Most tetrathyridia in the peritoneal cavity were free, but some were encapsulated in a thin connective tissue stroma and loosely attached to the inner organs. They appear as whitish, heart-shaped, flattened, unsegmented bodies with a slightly pointed posterior end. In the liver, tetrathyridia were seen as pale-tanned nodules embedded in the parenchyma. Comparative molecular analysis, both at the generic level (D1 domain LSU ribosomal DNA), and at the specific level (cytochrome c oxidase subunit I (cox1) and 12S mitochondrial DNA), revealed that the tetrathyridia belonged to M. canislagopodis. A. sylvaticus represents a new second intermediate host record in Iceland, and the first description of a rodent as intermediate host for this species, thus participating in the life cycle of the parasite.

Cystic echinococcosis in Iceland: a brief history and genetic analysis of a 46-year-old Echinococcus isolate collected prior to the eradication of this zoonotic disease.

Cystic echinococcosis (CE) is considered the most severe parasitic disease that ever affected the human population in Iceland. Before the start of eradication campaign in the 1860s, Iceland was a country with very high prevalence of human CE, with approximately every fifth person infected. Eradication of CE from Iceland by 1979 was a huge success story and served as a leading example for other countries on how to combat such a severe One Health problem. However, there is no genetic information on Echinococcus parasites before eradication. Here, we reveal the genetic identity for one of the last Echinococcus isolates in Iceland, obtained from a sheep 46 years ago (1977). We sequenced a large portion of the mitochondrial genome (8141 bp) and identified the isolate as Echinococcus granulosus sensu stricto genotype G1. As G1 is known to be highly infective genotype to humans, it may partly explain why such a large proportion of human population in Iceland was infected at a time . The study demonstrates that decades-old samples hold significant potential to uncover genetic identities of parasites in the past.

Prey-mimetism in cercariae of Apatemon (Digenea, Strigeidae) in freshwater in northern latitudes.

Cercariae, the free-living larval stages of trematodes, have adopted an amazing variety of transmission strategies. One of them is prey-mimetism, i.e. cercariae mimicking prey to attract motile hosts to be eaten. In a period between 2002 and 2019, we examined small planorbid snails, Bathyomphalus contortus, Gyraulus parvus and Planorbis planorbis from lakes in Finland and Iceland and from the Curonian Lagoon in Lithuania. Cercariae with conspicuously enlarged tails and unusual swimming behaviour, likely mimicking invertebrate prey, were detected and studied by the use of morphological and molecular (cox1, ITS1-5.8S-ITS2 and 28S rDNA) methods. Cercariae of two species belonging to the genus Apatemon (Strigeidae) were recognised. We consider Apatemon sp. 5 ex P. planorbis from the Curonian Lagoon identical to Cercaria globocaudata U. Szidat, 1940. Cercariae ex G. parvus from Iceland and ex B. contortus from Finland were conspecific, and we named them Apatemon sp. 6; these cercariae could not be associated with any known species. For the first time, we verified that cercariae of the Bulbocauda group belong to the genus Apatemon. We provide a mini-review on records of furcocercariae of the family Strigeidae with enlarged tails reported in freshwaters of the northern hemisphere and reveal that it is not only Apatemon but also Australapatemon and most likely Strigea which belong to the Bulbocauda group, rendering it a purely ecological assemblage. Understanding which invertebrate swimming behaviour these cercariae are mimicking will enhance our knowledge of the processes behind trematode transmission and will help to assess evolutionary pathways of host-finding strategies in trematodes.

Overview of renicolid digeneans (Digenea, Renicolidae) from marine gulls of northern Holarctic with remarks on their species statuses, phylogeny and phylogeography.

Renicolid digeneans parasitize aquatic birds. Their intramolluscan stages develop in marine and brackish-water gastropods, while metacercariae develop in molluscs and fishes. The systematics of renicolids is poorly developed, their life cycles are mostly unknown, and the statuses of many species require revision. Here, we establish based on integrated morphological and molecular data that adult renicolids from gulls Larus argentatus and Larus schistisagus and sporocysts and cercariae of Cercaria parvicaudata from marine snails Littorina spp. are life-cycle stages of the same species. We name it Renicola parvicaudatus and synonymized with it Renicola roscovitus. An analysis of the cox1 gene of R. parvicaudatus from Europe, North America and North Asia demonstrates a low genetic divergence, suggesting that this species has formed quite recently (perhaps during last glacial maximum) and that interregional gene flow is high. In Littorina saxatilis and L. obtusata from the Barents Sea, molecular analysis has revealed intramolluscan stages of Cercaria littorinae saxatilis VIII, a cryptic species relative to R. parvicaudatus. In the molecular trees, Renicola keimahuri from L. schistisagus belongs to another clade than R. parvicaudatus. We show that the species of this clade have cercariae of Rhodometopa group and outline morphological and behavioural transformations leading from xiphidiocercariae to these larvae. Molecular analysis has revealed 3 main phylogenetic branches of renicolids, differing in structure of adults, type of cercariae and host range. Our results elucidate the patterns of host colonization and geographical expansion of renicolids and pave the way to the solution of some long-standing problems of their classification.

Identifying sources of variation in parasite aggregation.

Aggregation of macroparasites among hosts is a near-universal pattern, and has important consequences for the stability of host-parasite associations and the impacts of disease. Identifying which potential drivers are contributing to levels of aggregation observed in parasite-host associations is challenging, particularly for observational studies. We apply beta regressions in a Bayesian framework to determine predictors of aggregation, quantified using Poulin's index of discrepancy (D), for 13 species of parasites infecting Icelandic Rock Ptarmigan (Lagopus muta) collected over 12 years. 1,140 ptarmigan were collected using sampling protocols maximizing consistency of sample sizes and of composition of host ages and sexes represented across years from 2006-2017. Parasite species, taxonomic group (insect, mite, coccidian, or nematode), and whether the parasite was an ecto- or endoparasite were tested as predictors of aggregation, either alone or by modulating an effect of parasite mean abundance on D. Parasite species was an important predictor of aggregation in models. Despite variation in D across samples and years, relatively consistent aggregation was demonstrated for each specific host-parasite association, but not for broader taxonomic groups, after taking sample mean abundance into account. Furthermore, sample mean abundance was consistently and inversely related to aggregation among the nine ectoparasites, however no relationship between mean abundance and aggregation was observed among the four endoparasites. We discuss sources of variation in observed aggregation, sources both statistical and biological in nature, and show that aggregation is predictable, and distinguishable, among infecting species. We propose explanations for observed patterns and call for the review and re-analysis of parasite and other symbiont distributions using beta regression to identify important drivers of aggregation-both broad and association-specific.

[Human Pseudoterranova and Anisakis cases in Iceland 2004-2020].

During 2004-2020 in total 18 anisakid larvae (Nematoda) were sent in to the Laboratory of Parasitology at Keldur for investigation and species identification. Fourteen had temporarily lived within the human body and were alive when detected, three were noticed alive in food just before being consumed, one larva was found dead. Pseudoterranova decipiens was found í 16 instances (89%), Anisakis simplex in two (11%). The one Anisakis case was a wriggling larva detected in the diaper of a baby that was believed to have ingested the larva with undercooked fish three days earlier in the kindergarten. In the other case a dead larva was found entangled in fish chew, spit out by a baby being fed with boiled haddock. Pseudoterranova larvae in humans (n=13) were most frequently detected in the mouth (11 persons). In one instance winding movements of larva in vomit of a baby attracted the attention of the mother, in another case a person detected tickling movements of a larva when cleaning the anal area after defecation. Length of the 13 Pseudoterranova larvae varied between 30 and 47 mm. They were believed to have lived in their hosts from one up to nine days. Nine larvae had already developed to the L4, stage, four were still in the L3 stage. Cod was the most frequently mentioned source of infection (5 of 14 cases), two persons regarded catfish to be the culprit, one named both fish species. In one case either sushi or plaice was believed to be the infection source, one person presumably got the larva participating in a sushi feast. In four cases the fish source remained unknown. Most often the larva was consumed in private homes, three persons believed to have gotten the larva when dining in a restaurant, a harbour worker got the worm when eating raw fish and the same baby got a larva on two different occasions in the kindergarten.

Dirofilaria spp. and Angiostrongylus vasorum: Current Risk of Spreading in Central and Northern Europe.

In the past few decades, the relevance of Dirofilaria immitis and Dirofilaria repens, causing cardiopulmonary and subcutaneous dirofilariosis in dogs and cats, and of Angiostrongylus vasorum, causing canine angiostrongylosis, has steadily increased in Central and Northern Europe. In this review, a summary of published articles and additional reports dealing with imported or autochthonous cases of these parasites is provided for Central (Austria, Czechia, Germany, Hungary, Luxemburg, Poland, Slovakia, Slovenia, and Switzerland) and Northern (Denmark, Finland, Iceland, Norway, and Sweden) Europe. Research efforts focusing on Dirofilaria spp. and A. vasorum have varied by country, and cross-border studies are few. The housing conditions of dogs, pet movements, the spread of competent vectors, and climate change are important factors in the spread of these nematodes. Dogs kept outside overnight are a major factor for the establishment of Dirofilaria spp. However, the establishment of invasive, diurnal, synanthropic, competent mosquito vectors such as Aedes albopictus may also influence the establishment of Dirofilaria spp. The drivers of the spread of A. vasorum remain not fully understood, but it seems to be influenced by habitats shared with wild canids, dog relocation, and possibly climatic changes; its pattern of spreading appears to be similar in different countries. Both Dirofilaria spp. and A. vasorum merit further monitoring and research focus in Europe.

Diversity of echinostomes (Digenea: Echinostomatidae) in their snail hosts at high latitudes.

The biodiversity of freshwater ecosystems globally still leaves much to be discovered, not least in the trematode parasite fauna they support. Echinostome trematode parasites have complex, multiple-host life-cycles, often involving migratory bird definitive hosts, thus leading to widespread distributions. Here, we examined the echinostome diversity in freshwater ecosystems at high latitude locations in Iceland, Finland, Ireland and Alaska (USA). We report 14 echinostome species identified morphologically and molecularly from analyses of nad1 and 28S rDNA sequence data. We found echinostomes parasitising snails of 11 species from the families Lymnaeidae, Planorbidae, Physidae and Valvatidae. The number of echinostome species in different hosts did not vary greatly and ranged from one to three species. Of these 14 trematode species, we discovered four species (Echinoparyphium sp. 1, Echinoparyphium sp. 2, Neopetasiger sp. 5, and Echinostomatidae gen. sp.) as novel in Europe; we provide descriptions for the newly recorded species and those not previously associated with DNA sequences. Two species from Iceland (Neopetasiger islandicus and Echinoparyphium sp. 2) were recorded in both Iceland and North America. All species found in Ireland are new records for this country. Via an integrative taxonomic approach taken, both morphological and molecular data are provided for comparison with future studies to elucidate many of the unknown parasite life cycles and transmission routes. Our reports of species distributions spanning Europe and North America highlight the need for parasite biodiversity assessments across large geographical areas.

TITLE: Diversité des Échinostomes (Digenea, Echinostomatidae) chez leurs hôtes mollusques aux latitudes élevées. ABSTRACT: La biodiversité des écosystèmes d'eau douce à l'échelle mondiale laisse encore beaucoup à découvrir, notamment dans la faune parasitaire des trématodes qu'ils abritent. Les parasites trématodes Échinostomes ont des cycles de vie complexes à hôtes multiples impliquant souvent des oiseaux migrateurs comme hôtes définitifs, conduisant ainsi à des distributions étendues. Ici, nous avons examiné la diversité des échinostomes dans les écosystèmes d'eau douce à des latitudes élevées en Islande, Finlande, Irlande et en Alaska (États-Unis). Nous rapportons de séquences de nad1 et d'ADNr 28S morphologiquement et moléculairement à partir d'analyses de données de séquence d'ADNr nad1 et 28S. Nous avons trouvé des échinostomes parasitant les mollusques de 11 espèces des familles Lymnaeidae, Planorbidae, Physidae et Valvatidae. Le nombre d'espèces d'échinostomes dans différents hôtes ne variait pas beaucoup et allait d'une à trois espèces. Sur ces 14 espèces de trématodes, nous avons découvert quatre espèces (Echinoparyphium sp. 1, Echinoparyphium sp. 2, Neopetasiger sp. 5, Echinostomatidae gen. sp.) comme nouvelles pour l'Europe; nous fournissons des descriptions pour les espèces nouvellement signalées et celles qui n'étaient pas précédemment associées à des séquences d'ADN. Deux espèces d'Islande (Neopetasiger islandicus et Echinoparyphium sp. 2) ont été signalées en Islande et en Amérique du Nord. Toutes les espèces trouvées en Irlande sont de nouveaux signalements pour ce pays. Grâce à une approche taxonomique intégrative, des données morphologiques et moléculaires sont fournies à des fins de comparaison avec des études futures afin d'élucider les nombreux cycles de vie et voies de transmission des parasites, qui sont inconnus. Nos données sur la répartition des espèces en Europe et en Amérique du Nord soulignent la nécessité d'évaluer la biodiversité des parasites dans de vastes zones géographiques.

Post-Translational Protein Deimination Signatures in Plasma and Plasma EVs of Reindeer (Rangifer tarandus).

The reindeer (caribou) Rangifer tarandus is a Cervidae in the order Artiodactyla. Reindeer are sedentary and migratory populations with circumpolar distribution in the Arctic, Northern Europe, Siberia and North America. Reindeer are an important wild and domesticated species, and have developed various adaptive strategies to extreme environments. Importantly, deer have also been identified to be putative zoonotic carriers, including for parasites, prions and coronavirus. Therefore, novel insights into immune-related markers are of considerable interest. Peptidylarginine deiminases (PADs) are a phylogenetically conserved enzyme family which causes post-translational protein deimination by converting arginine into citrulline in target proteins. This affects protein function in health and disease. Extracellular vesicles (EVs) participate in cellular communication, in physiological and pathological processes, via transfer of cargo material, and their release is partly regulated by PADs. This study assessed deiminated protein and EV profile signatures in plasma from sixteen healthy wild female reindeer, collected in Iceland during screening for parasites and chronic wasting disease. Reindeer plasma EV profiles showed a poly-dispersed distribution from 30 to 400 nm and were positive for phylogenetically conserved EV-specific markers. Deiminated proteins were isolated from whole plasma and plasma EVs, identified by proteomic analysis and protein interaction networks assessed by KEGG and GO analysis. This revealed a large number of deimination-enriched pathways for immunity and metabolism, with some differences between whole plasma and EVs. While shared KEGG pathways for whole plasma and plasma EVs included complement and coagulation pathways, KEGG pathways specific for EVs were for protein digestion and absorption, platelet activation, amoebiasis, the AGE-RAGE signaling pathway in diabetic complications, ECM receptor interaction, the relaxin signaling pathway and the estrogen signaling pathway. KEGG pathways specific for whole plasma were pertussis, ferroptosis, SLE, thyroid hormone synthesis, phagosome, Staphylococcus aureus infection, vitamin digestion and absorption, and prion disease. Further differences were also found between molecular function and biological processes GO pathways when comparing functional STRING networks for deiminated proteins in EVs, compared with deiminated proteins in whole plasma. This study highlights deiminated proteins and EVs as candidate biomarkers for reindeer health and may provide information on regulation of immune pathways in physiological and pathological processes, including neurodegenerative (prion) disease and zoonosis.

Development of 14 Microsatellite Markers for Zoonotic Tapeworm Dibothriocephalus dendriticus (Cestoda: Diphyllobothriidea).

Dibothriocephalus dendriticus is one of the causative agents of the fish-borne zoonosis diphyllobothriosis. Polymorphic microsatellite markers were originally developed for future genetic studies using microsatellite library screening and next-generation sequencing (NGS). Out of 128 microsatellite candidates selected after NGS analysis, 126 yielded PCR products of the expected size. A declared repetitive motif was confirmed in 92 loci by Sanger sequencing. The level of polymorphism was tested by fragment analysis. Statistical tests for observed and expected heterozygosities and deviations from Hardy-Weinberg equilibrium revealed 14 polymorphic microsatellite loci suitable for studies on the finer genetic structure of global populations of D. dendriticus.

Presence of eimerid oocysts in faeces of a quarantined dog in Iceland is explained by coprophagic behaviour prior to its importation. Case report.

BACKGROUND: All dogs imported into Iceland must undergo mandatory quarantine in a special station before introduction into the country. A faecal sample is collected from the first stool passed by the dog in this station and subsequently examined for the presence of intestinal parasite stages. CASE PRESENTATION: In May 2019 unsporulated oocysts were detected in faeces from a 7-year-old household dog that had been imported from Sweden. Most of the oocysts studied strongly resembled those of Eimeria canis Wenyon, 1923. As this species is not valid, the purpose of the present article was to identify the correct species and examine their possible origin. Studies confirmed the presence of two distinct unsporulated oocyst morphotypes in the faeces; measurements and photomicrographs confirmed their identification as Eimeria magna Pérard, 1925 and Eimeria stiedai (Lindemann, 1865) Kisskalt and Hartmann, 1907, both common parasites of European rabbits, Oryctolagus cuniculus (L., 1758). When the owner of the dog was questioned about the food administrated to the dog prior to its import to Iceland, it turned out that it had exclusively been fed dry dog food pellets. However, the owner also reported that on the morning prior to transportation to Iceland, the dog was allowed to move freely in a grassland area where rabbits are common and heaps of their faeces are present. Furthermore, the owner confirmed that the dog consumed rabbit faeces that morning. CONCLUSION: It is believed that this coprophagic behaviour can explain the detection of rabbit eimerids in the dog's faeces, and that such behaviour must be taken into consideration by veterinarians and other diagnostic personnel when they detect atypical cysts or eggs during coprological examinations.

Deiminated proteins and extracellular vesicles as novel biomarkers in pinnipeds: Grey seal (Halichoerus gryptus) and harbour seal (Phoca vitulina).

Peptidylarginine deiminases (PADs) are phylogenetically conserved calcium-dependent enzymes which post-translationally convert arginine into citrulline in target proteins in an irreversible manner, leading to functional and structural changes in target proteins. Protein deimination can cause the generation of neo-epitopes, affect gene regulation and also allow for protein moonlighting and therefore facilitate multifaceted functions of the same protein. PADs are furthermore a key regulator of cellular release of extracellular vesicle (EVs), which are found in most body fluids and participate in cellular communication via transfer of cargo proteins and genetic material. In this study, post-translationally deiminated proteins and EVs were assessed in sera of two seal species, grey seal and harbour seal. We report a poly-dispersed population of serum-EVs, which were positive for phylogenetically conserved EV-specific markers and characterised by transmission electron microscopy. A number of deiminated proteins critical for immune and metabolic functions were identified in the seal sera and varied somewhat between the two species under study, while some targets were in common. EV profiles of the seal sera further revealed that key microRNAs for inflammation, immunity and hypoxia also vary between the two species. Protein deimination and EVs profiles may be useful biomarkers for assessing health status of sea mammals, which face environmental challenges, including opportunistic infection, pollution and shifting habitat due to global warming.

Quill mite infestation of rock ptarmigan Lagopus muta (Aves: Phasianidae) in relation to year and host age, sex, body condition, and density.

A total of 1209 ptarmigan were examined for Mironovia lagopus, including 721 juvenile birds (ca. 3 months old) and 488 adult birds (15 months or older). A total of 88 birds or 7.3% (n = 1209, 95% cl 5.9-8.9%) were infested with M. lagopus. There was an age difference in prevalence of infection, and more adults (10.7%, n = 488, 95% cl 8.2-13.7%) than juveniles (5.0%, n = 721, 95% cl 3.6-6.8%) were infested. There was a significant age effect in the mean intensity index, and adult birds had more advanced infestations compared with juvenile birds. There were no significant changes in either the interannual prevalence of infection or the mean intensity index of infection. Of the feather types inspected, there was no age-related difference in selection of feathers, nor was there any preference of mites for any one of the inspected feather types. Body dispersants were all adult females. There was an age-related difference in mean intensities of infection of body dispersants; it was higher in adult birds. The methods used to determine presence or absence of M. lagopus were not 100% accurate especially for juvenile hosts, and this at least partly explains the difference in prevalence among age groups. There was no relationship between host body condition or host density and infection by M. lagopus.

Transatlantic discovery of Notocotylus atlanticus (Digenea: Notocotylidae) based on life cycle data.

Digenean parasites feature a series of stages with a distinct appearance, reproduction mode, and lifestyle that together constitute their well-known, complex life cycle. Species descriptions of Digenea have always been based on one of these stages-the marita, or sexually reproducing adult in the final host. However, in some cases, data on the life cycle are essential for the differential diagnosis of closely related species. Here, we present the case of Notocotylus atlanticus, where different stages of its life cycle were discovered for the first time since the species description, and across the Atlantic. We used a material from a naturally infected intertidal marine snail, Ecrobia ventrosa, and several waterfowl species and also carried out infection experiments. For morphological studies, we employed light microscopy, SEM, and CLSM; molecular data obtained include sequences of ITS1 and 28S rRNA gene. We demonstrate that N. atlanticus adult worm morphology is barely sufficient to distinguish it from several other species. Cercariae morphology and identity of the first intermediate hosts provide crucial additional information. According to our preliminary phylogenetic reconstructions, two notocotylid lineages are associated with two major gastropod lineages-the Caenogastropoda and the Heterobranchia. The traditional character to identify notocotylid genera (structure of ventral organs) fails to explain the phylogeny and thus requires reassessment. Further reliable morphological, life cycle and molecular data on other species are likely to reveal more patterns in notocotylid systematics, host specificity, and evolution.

Analysis and Characterization of Polychlorinated Hydroxybornanes as Metabolites of Toxaphene Using a Polar Bear Model.

Abiotic and biotic transformation of toxaphene (camphechlor) results in the selective enrichment of recalcitrant congeners while other, less persistent compounds of technical toxaphene (CTTs) are degraded. Until now, there has been little knowledge on oxidation transformation of toxaphene. For instance, the existence of hydroxylated CTTs (OH-CTTs) in authentic environmental and food samples has not been proven. For this reason, we synthesized a mixture consisting of tetra- to heptachlorinated OH-CTTs and simplified it by countercurrent chromatography (CCC). Thus, 227 OH-CTTs were detected in the CCC fractions (12 tetra-, 117 penta-, 81 hexa-, and 17 heptachlorinated OH-CTTs), which was >50% more than detected before the fractionation. One CCC fraction consisting of only 18 OH-CTTs was used to develop a sample cleanup method which aimed to remove CTTs, isobaric PCBs, and sample matrix. The final cleanup procedure consisted of (i) gel permeation chromatography (GPC) and adsorption chromatography using (ii) deactivated and (iii) activated silica gel. Hence, up to 320 and 4350 µg/kg lipid weight of octa- and nonachlorinated CTTs were detected in four liver samples and adipose tissue of polar bears, respectively. Furthermore, the presence of one hexachlorinated OH-CTT isomer could be verified in the samples, which was about 1% of the octachlorinated CTTs determined in the liver samples.

Combined thin layer chromatography and gas chromatography with mass spectrometric analysis of lipid classes and fatty acids in malnourished polar bears (Ursus maritimus) which swam to Iceland.

Between 2008 and 2011, four polar bears (Ursus maritimus) from the Greenland population swam and/or drifted on ice to Iceland where they arrived in very poor body condition. Body fat resources in these animals were only between 0% and 10% of the body weight (usually 25%). Here we studied the lipid composition in different tissues (adipose tissue if available, liver, kidney and muscle). Lipid classes were determined by thin layer chromatography (TLC) and on-column gas chromatography with mass spectrometry (GC/MS). The fatty acid pattern of total lipids and free fatty acids was analyzed by GC/MS in selected ion monitoring (SIM) mode. Additionally, cholesteryl esters and native fatty acid methyl esters, initially detected as zones in thin layer chromatograms, were enriched by solid phase extraction and quantified by GC/MS. The ratio of free fatty acids to native fatty acid methyl esters could be correlated with the remained body lipids in the polar bears and thus may also serve as a marker for other starving animals or even for humans.

Host-Parasite Interactions and Population Dynamics of Rock Ptarmigan.

Populations of rock ptarmigan (Lagopus muta) in Iceland fluctuate in multiannual cycles with peak numbers c. every 10 years. We studied the ptarmigan-parasite community and how parasites relate to ptarmigan age, body condition, and population density. We collected 632 ptarmigan in northeast Iceland in early October from 2006 to 2012; 630 (99.7%) were infected with at least one parasite species, 616 (98%) with ectoparasites, and 536 (85%) with endoparasites. We analysed indices for the combined parasite community (16 species) and known pathogenic parasites, two coccidian protozoans Eimeria muta and Eimeria rjupa, two nematodes Capillaria caudinflata and Trichostrongylus tenuis, one chewing louse Amyrsidea lagopi, and one skin mite Metamicrolichus islandicus. Juveniles overall had more ectoparasites than adults, but endoparasite levels were similar in both groups. Ptarmigan population density was associated with endoparasites, and in particular prevalence of the coccidian parasite Eimeria muta. Annual aggregation level of this eimerid fluctuated inversely with prevalence, with lows at prevalence peak and vice versa. Both prevalence and aggregation of E. muta tracked ptarmigan population density with a 1.5 year time lag. The time lag could be explained by the host specificity of this eimerid, host density dependent shedding of oocysts, and their persistence in the environment from one year to the next. Ptarmigan body condition was negatively associated with E. muta prevalence, an indication of their pathogenicity, and this eimerid was also positively associated with ptarmigan mortality and marginally inversely with fecundity. There were also significant associations between fecundity and chewing louse Amyrsidea lagopi prevalence (negative), excess juvenile mortality and nematode Capillaria caudinflata prevalence (positive), and adult mortality and skin mite Metamicrolichus islandicus prevalence (negative). Though this study is correlational, it provides strong evidence that E. muta through time-lag in prevalence with respect to host population size and by showing significant relations with host body condition, mortality, and fecundity could destabilize ptarmigan population dynamics in Iceland.

Morphological characteristics of Mesocestoides canislagopodis (Krabbe 1865) tetrathyridia found in rock ptarmigan (Lagopus muta) in Iceland.

Necropsies of 1010 rock ptarmigans (Lagopus muta) sampled in autumn 2006-2015 in northeast Iceland revealed Mesocestoides canislagopodis tetrathyridia infections in six birds (0.6 %), two juvenile birds (3 month old), and four adult birds (15 months or older). Four birds had tetrathyridia in the body cavity, one bird in the liver, and one bird both in the body cavity and the liver. There were more tetrathyridia in the body cavity of the two juveniles (c. 50 in each) than in three adults (10-40), possibly indicating a host-age-related tetrathyridia mortality. Approximately, half of tetrathyridia in the body cavity were free or loosely attached to the serosa, the other half were encapsulated in a thin, loose connective tissue stroma, frequently attached to the lungs and the liver. Tetrathyridia in the liver parenchyma incited variably intense inflammation. Tetrathyridia from the juvenile hosts were whitish, heart-shaped, and flattened, with unsegmented bodies with a slightly pointed posterior end. In the adult hosts, tetrathyridia were sometimes almost rectangular-shaped, slightly wider compared to those in the juveniles, but more than twice as long as the younger-aged tetrathyridia. Tetrathyridia infections are most likely acquired during the brief insectivorous feeding phase of ptarmigan chicks, and the tetrathyridia persist throughout the lifespan of the birds.