Matthías Eydal

PARASITES OF THE ARCTIC FOX (ALOPEX LAGOPUS) IN ICELAND

Forty-four of 50 arctic foxes (Alopex lagopus) in Iceland harbored 15 species of intestinal parasites, including Protozoa: Eimeria sp. or Isospora sp. (in 4%); Trematoda: Cryptocotyle lingua (24%), Plagiorchis elegans (4%), Brachylaemus sp. (12%), Tristriata sp. (10%), and Spelotrema sp. (8%); Cestoda: Mesocestoides canislagopodis (72%), Schistocephalus solidus (2%), and Diphyllobothrium dendriticum (4%); Nematoda: Toxascaris leonina (50%), Toxocara canis (2%), Uncinaria stenocephala (4%), and eggs of the lung worm Capillaria aerophila (6%); and Acanthocephala: Polymorphus meyeri (8%) and Corynosoma hadweni (2%). Only four of the species previously had been recorded in Iceland. Eleven species are new records in Iceland and six appear to be new host records. Two additional nematodes, Stegophorus stercorarii and Syphacia sp., probably were ingested accidentally with the prey. Foxes from coastal habitats harbored 14 parasitic species while only five species were found in foxes from inland habitats. Arctic foxes from coastal habitats generally had higher helminth burdens and harbored more parasitic species per fox than foxes from inland habitats.

Fatal infection in two Icelandic stallions caused by Halicephalobus gingivalis (Nematoda: Rhabditida)

Opportunistic infections with the free living nematode Halicephalobus gingivalis are infrequently reported in horses but the cases are widespread geographically. The nematodes are believed to penetrate wounds and subsequently reproduce within the host tissues. This paper reports two cases of a fatal disease in stallions of the Icelandic breed in Iceland. Case 1: a stallion, which sustained injuries to the mouth after an accident, developed severe neurological signs and had to be euthanatized. Histological examination revealed mild inflammation and malacia in the cerebellum associated with the presence of numerous H. gingivalis nematodes. Case 2: a stallion that started swerving to one side and lost balance was euthanatized due to lack of response to therapy and rapid deterioration. Histological examination revealed numerous H. gingivalis nematodes in the cerebellum, brain stem, cervical spinal cord and in the meninges, with minimal reactive changes. In case 1 the infection presumably was acquired by nematodes from soil penetrating through wounds in the mouth. The mode of the H. gingivalis infection in case 2 is uncertain. These are the first cases of H. gingivalis infection reported from Iceland and the second report from the Nordic countries.

Previously unknown apicomplexan species infecting Iceland scallop, Chlamys islandica (Müller, 1776), queen scallop, Aequipecten opercularis L., and king scallop, Pecten maximus L.

Examination of three scallop species from three separate locations: Iceland scallop from Icelandic waters, king scallop from Scottish waters and queen scallop from Faroese and Scottish waters, revealed infections of a previously unknown apicomplexan parasite in all three scallop species. Developmental forms observed in the shells appeared to include both sexual and asexual stages of the parasite, i.e. merogony, gametogony and sporogony, which suggests a monoxenous life cycle. Meronts, gamonts, zygotes and mature oocysts were solely found in the muscular tissue. Zoites, which could be sporozoites and/or merozoites, were observed in great numbers, most frequently in muscles, both intracellular and free in the extracellular space. Zoites were also common inside haemocytes. Examination of the ultrastructure showed that the zoites contained all the major structures characterizing apicomplexans. This apicomplexan parasite is morphologically different from other apicomplexan species previously described from bivalves. Presently, its systematic position within the phylum Apicomplexa cannot be ascertained.

Margolisiella islandica sp. nov. (Apicomplexa: Eimeridae) infecting Iceland scallop Chlamys islandica (Müller, 1776) in Icelandic waters

Wild Iceland scallops Chlamys islandica from an Icelandic bay were examined for parasites. Queen scallops Aequipecten opercularis from the Faroe Islands and king scallops Pecten maximus and queen scallops from Scottish waters were also examined. Observations revealed heavy infections of eimeriorine parasites in 95-100% of C. islandica but not the other scallop species. All life stages in the apicomplexan reproduction phases, i.e. merogony, gametogony and sporogony, were present. Trophozoites and meronts were common within endothelial cells of the hearts auricle and two generations of free merozoites were frequently seen in great numbers in the haemolymph. Gamonts at various developmental stages were also abundant, most frequently free in the haemolymph. Macrogamonts were much more numerous than microgamonts. Oocysts were exclusively in the haemolymph; live mature oocysts contained numerous (>500) densely packed pairs of sporozoites forming sporocysts. Analysis of the 18S ribosomal DNA revealed that the parasite from C. islandica is most similar (97.7% identity) to an unidentified apicomplexan isolated from the haemolymph of the giant clam, Tridacna crocea, from Japan. Phylogenetic analyses showed that the novel sequence consistently grouped with the Tridacna sequence which formed a robust sister clade to the rhytidocystid group. We propose the name Margolisiella islandica sp. nov., referring to both type host and type locality.

Molecular identification and transmission studies of X-cell parasites from Atlantic cod Gadus morhua (Gadiformes: Gadidae) and the northern black flounder Pseudopleuronectes obscurus (Pleuronectiformes: Pleuronectidae)

BackgroundEpidermal pseudotumours from Hippoglossoides dubius and Acanthogobius flavimanus in Japan and gill lesions in Limanda limanda from the UK have been shown to be caused by phylogenetically related protozoan parasites, known collectively as X-cells. However, the phylogenetic position of the X-cell group is not well supported within any of the existing protozoan phyla and they are currently thought to be members of the Alveolata.Ultrastructural features of X-cells in fish pseudotumours are somewhat limited and no typical environmental stages, such as spores or flagellated cells, have been observed. The life cycles for these parasites have not been demonstrated and it remains unknown how transmission to a new host occurs.In the present study, pseudobranchial pseudotumours from Atlantic cod, Gadus morhua, in Iceland and epidermal pseudotumours from the northern black flounder, Pseudopleuronectes obscurus, in Japan were used in experimental transmission studies to establish whether direct transmission of the parasite is achievable. In addition, X-cells from Atlantic cod were sequenced to confirm whether they are phylogenetically related to other X-cells and epidermal pseudotumours from the northern black flounder were analysed to establish whether the same parasite is responsible for infecting different flatfish species in Japan.ResultsPhylogenetic analyses of small subunit ribosomal DNA (SSU rDNA) sequence data from Atlantic cod X-cells show that they are a related parasite that occupies a basal position to the clade containing other X-cell parasites. The X-cell parasite causing epidermal pseudotumours in P. obscurus is the same parasite that causes pseudotumours in H. dubius. Direct, fish to fish, transmission of the X-cell parasites used in this study, via oral feeding or injection, was not achieved. Non-amoeboid X-cells are contained within discrete sac-like structures that are loosely attached to epidermal pseudotumours in flatfish; these X-cells are able to tolerate exposure to seawater.A sensitive nested PCR assay was developed for the sub clinical detection of both parasites and to assist in future life cycle studies. PCR revealed that the parasite in P. obscurus was detectable in non-pseudotumourous areas of fish that had pseudotumours present in other areas of the body.ConclusionsThe inability to successfully transmit both parasites in this study suggests that either host detachment combined with a period of independent development or an alternate host is required to complete the life cycle for X-cell parasites. Phylogenetic analyses of SSU rDNA confirm a monophyletic grouping for all sequenced X-cell parasites, but do not robustly support their placement within any established protist phylum. Analysis of SSU rDNA from X-cells in Japanese flatfish reveals that the same parasite can infect more than one species of fish.

Prosorhynchoides borealis Bartoli, Gibson & Bray, 2006 (Digenea: Bucephalidae) cercariae from Abra prismatica (Mollusca: Bivalvia) in Icelandic waters

This paper reports the adult stage of Prosorhynchoides borealis (Digenea) from Lophius piscatorius in Icelandic waters and infections with the larval stages (sporocysts and cercariae) found for the first time in the bivalve Abra prismatica (Semelidae). The previously known first intermediate host was Abra alba (Semelidae). Ribosomal DNA sequencing studies on all three life stages of the parasite (cercariae, metacercariae, adults) were performed to confirm their identites. Morphometric measurements confirmed that the adult worms belong to the newly described species P. borealis. Prosorhynchoides borealis sporocysts filled with cercariae were found in 16% of A. prismatica bivalves sampled at depths between 34 and 93 m off South Iceland. Prevalence ranged from 0 to 44% between different localities. The parasite was found only in the larger bivalves. Extensive sporocyst infection in the haemocoel of the foot caused mechanical muscle damage with subsequent degeneration and necrosis. Other tissues, including the digestive gland, nephridia, gills and intestine, were less heavily infected. Only focal necrosis was observed in the digestive gland, nephridia and gills, and local atrophy in the intestine. Cercariae were also observed in the lumen of both the stomach and intestine. This is the first report of A. prismatica as an alternative first intermediate host for P. borealis. Ribosomal DNA sequence data reveals 100% homology in the data between cercariae, metacercariae and adult digeneans, supporting the morphological data suggesting that all stages belong to the same species.

Pseudobranchial X-cell pseudotumors in young wild and farmed Atlantic cod Gadus morhua in Iceland

This study investigated the prevalence of pseudobranchial X-cell pseudotumors in 2 year classes of wild juvenile cod and to a lesser extent in older cod in NW Iceland. The disease was also monitored in farmed wild-caught juvenile cod. The youngest wild cod detected that had X-cell infections were 6.5 to 13.0 cm in length, with prevalence reaching a maximum level of 7%. The highest prevalence (23%) was recorded in 18.5 to 27.0 cm fish. The mean prevalence in older cod (25.0 to 76.0 cm in length) was 7% but decreased with increasing age. The mean prevalence of X-cell pseudotumors in farmed juvenile cod was < or =1% during winter rearing in land-based tanks. Higher prevalence (2 to 15%) was noted in dead and moribund fish. Diseased farmed fish were usually emaciated, and mortality due to the disease was noticeable.

A Recent Whole-Genome Duplication Divides Populations of a Globally Distributed Microsporidian

The Microsporidia are a major group of intracellular fungi and important parasites of animals including insects, fish, and immunocompromised humans. Microsporidian genomes have undergone extreme reductive evolution but there are major differences in genome size and structure within the group: some are prokaryote-like in size and organisation (<3 Mb of gene-dense sequence) while others have more typically eukaryotic genome architectures. To gain fine-scale, population-level insight into the evolutionary dynamics of these tiny eukaryotic genomes, we performed the broadest microsporidian population genomic study to date, sequencing geographically isolated strains of Spraguea, a marine microsporidian infecting goosefish worldwide. Our analysis revealed that population structure across the Atlantic Ocean is associated with a conserved difference in ploidy, with American and Canadian isolates sharing an ancestral whole genome duplication that was followed by widespread pseudogenisation and sorting-out of paralogue pairs. While past analyses have suggested de novo gene formation of microsporidian-specific genes, we found evidence for the origin of new genes from noncoding sequence since the divergence of these populations. Some of these genes experience selective constraint, suggesting the evolution of new functions and local host adaptation. Combining our data with published microsporidian genomes, we show that nucleotide composition across the phylum is shaped by a mutational bias favoring A and T nucleotides, which is opposed by an evolutionary force favoring an increase in genomic GC content. This study reveals ongoing dramatic reorganization of genome structure and the evolution of new gene functions in modern microsporidians despite extensive genomic streamlining in their common ancestor.

Strongyloides stercoralis found in imported dogs, household dogs and kennel dogs in Iceland

The aim of this paper is to report cases of the intestinal parasitic nematode Strongyloides stercoralis in dogs in Iceland. The nematode was diagnosed in 20 (0.6%) of imported dogs in quaratine in Iceland during 19892016. Household dogs: The first case of S. stercoralis infection diagnosed outside quaratine in Iceland was in an unhealthy household puppy purchased from an Icelandic breeding kennel (Kennel A) in 2012. A total of nine puppies purchased from Kennel A, and two dogs which had contact with dogs from the kennel, were diagnosed with S. stercoralis. Kennel dogs: In 2012 S. stercoralis was confirmed in dozens of dogs in Kennel A. Follow-up examinations after anthelmintic treatments indicated a successful removal of worms in imported and household dogs. In spite of more than a dozen anthelmintic treatment actions and other arrangements in Kennel A since 2012, recurrent infections have repeatedly been confirmed, the last one in 2015. The nematode is believed to have been introduced to the breeding kennel with an imported dog, in spite of anthelmintic treatments in quarantine. YFIRLIT Sníkjuþráðormurinn Strongyloides stercoralis staðfestur á Íslandi í innfluttum hundum, heimilishundum og í hundaræktunarstöð. Markmið þessa yfirlits er að greina frá tilfellum sem greinst hafa af sníkjuþráðorminum Strongyloides stercoralis í hundum á Íslandi. Ormurinn hefur greinst í saursýnum úr 20 (0.6%) hundum sem fluttir voru til landins um einungrunarstöðvar á árunum 1989-2016. Heimilishundar: Fyrsta tilfelli utan einangrunarstöðva greindist í veikum heimilishvolpi sem keyptur var á íslenskri hundaræktunarstöð 2012. Ormurinn greindist í alls níu hvolpum sem keyptir voru á stöðinni og í tveimur heimilishundum sem höfðu haft samgang við hunda frá stöðinni. Hundaræktunarstöðin: S. stercoralis greindist í tugum hunda í stöðinni í byrjun árs 2012, bæði í hvolpum og fullorðnum hundum. Innfluttu hundarnir og heimilishundarnir fengu ormalyfjameðferð og eftirfylgnirannsóknir bentu til að tekist hefði að uppræta ormana. Margendurteknar ormalyfjagjafir á árunum 2012 -2016 og aðrar aðgerðir í hundaræktunarstöðinni virðast hafa borið umtalsverðan árangur en ormurinn hefur þó greinst í stökum saursýnum á undanförnum árum, síðast í nóvember 2015. Talið er að þráðormurinn hafi borist inn í hundaræktunarstöðina með innfluttum hundi, þrátt fyrir endurteknar ormalyfjagjafir í einangrunarstöð, smitast milli hunda í ræktunarstöðinni og borist þaðan út með seldum hundum.

Integrative Approach for the Reliable Detection and Specific Identification of the Microsporidium Loma morhua in Atlantic Cod (Gadus morhua)

Microsporidia are fungal parasites that infect diverse invertebrate and vertebrate hosts. Finfish aquaculture supports epizootics due to high host density and the high biotic potential of these parasites. Reliable methods for parasite detection and identification are a necessary precursor to empirical assessment of strategies to mitigate the effects of these pathogens during aquaculture. We developed an integrative approach to detect and identify Loma morhua infecting Atlantic cod. We show that the spleen is more reliable than the commonly presumed gills as best organ for parasite detection in spite of substantial morphological plasticity in xenoma complexes. We developed rDNA primers with 100% sensitivity in detecting L. morhua and with utility in distinguishing some congeneric Loma species. ITS sequencing is necessary to distinguish L. morhua from other congeneric microsporidia due to intraspecific nucleotide variation. 64% of L. morhua ITS variants from Atlantic cod have a 9‐nucleotide motif that distinguishes it from Loma spp. infecting non‐Gadus hosts. The remaining 36% of ITS variants from Atlantic cod are distinguished from currently represented Loma spp., particularly those infecting Gadus hosts, based on a 14‐nucleotide motif. This research approach is amenable to developing templates in support of reliable detection and identification of other microsporidian parasites in fishes.