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.

Molecular Characterization of Mesocestoides litteratus (Cestoda: Cyclophyllidea: Mesocestoididae) Tetrathyridium Isolated from Two Species of Rodents from Iran.

PURPOSE: Mesocestoides spp. are Cyclophyllidean tapeworms with zoonotic importance. The current study aimed to investigate the molecular characteristics of Mesocestoides larvae (tetrathyridium) isolated from the abdominal cavity of persion jird, Meriones persicus, and from the liver of grey hamster, Cricetulus migratorius, in Ardabil Province, northwest Iran. METHODS: Genomic DNA of the isolates of Mesocestoides tetrathyridium were extracted, and mitochondrial gene of cytochrome-c oxidase subunit1 (cox1) was amplified. Sequencing of PCR products were performed and phylogenic analysis was run using MEGA 6.0 software. RESULTS: Both isolates were identified as Mesocestoides litteratus, showing high identity with M. litteratus sequences available in GenBank. Also, they had 100% homology to each other. Intra-species variation within isolates of M. litteratus were 0-2.4%. The phylogenetic reconstruction based on the partial sequence of the cox1 gene showed that our sequences of M. litteratus were clustered with M. litteratus isolates from Slovakia, Netherlands, Germany and Italy. CONCLUSION: This is the first molecular description of M. litteratus from M. persicus and C. migratorius. Phylogenetic analysis illustrated that M. litteratus isolates of the current study had very high identities with the isolates of this species from other countries.

Cell repertoire and proliferation of germinative cells of the model cestode Mesocestoides corti.

The phylum Platyhelminthes shares a unique population of undifferentiated cells responsible for the proliferation capacity needed for cell renewal, growth, tissue repair and regeneration. These cells have been extensively studied in free-living flatworms, whereas in cestodes the presence of a set of undifferentiated cells, known as germinative cells, has been demonstrated in classical morphology studies, but poorly characterized with molecular biology approaches. Furthermore, several genes have been identified as neoblast markers in free-living flatworms that deserve study in cestode models. Here, different cell types of the model cestode Mesocestoides corti were characterized, identifying differentiated and germinative cells. Muscle cells, tegumental cells, calcareous corpuscle precursor cells and excretory system cells were identified, all of which are non-proliferative, differentiated cell types. Besides those, germinative cells were identified as a population of small cells with proliferative capacity in vivo. Primary cell culture experiments in Dulbecco's Modified Eagle Medium (DMEM), Echinococcus hydatid fluid and hepatocyte conditioned media in non-reductive or reductive conditions confirmed that the germinative cells were the only ones with proliferative capacity. Since several genes have been identified as markers of undifferentiated neoblast cells in free-living flatworms, the expression of pumilio and pL10 genes was analysed by qPCR and in situ hybridization, showing that the expression of these genes was stronger in germinative cells but not restricted to this cell type. This study provides the first tools to analyse and further characterise undifferentiated cells in a model cestode.

Global prevalence of Mesocestoides infections in animals - A systematic review and meta-analysis.

Mesocestoides spp. are zoonotic cestodes found as adults in carnivorous domestic and wild definitive hosts and as metacestodes in several taxa of intermediate hosts. Although several regional studies record its occurrence in different host populations, the global prevalence and patterns of occurrence of Mesocestoides spp. are not fully understood. The objective of this study was to conduct a systematic review and meta-analysis of published literature to estimate the global prevalence of Mesocestoides spp. in major definitive and intermediate host taxa. Records published in English were collected from NCBI PubMed, Science Direct, Web of Science and Google Scholar databases, with 364 papers being included in the meta-analysis. The overall pooled prevalence estimates show that 21.72 % (95 % CI: 18.49-25.14) of terrestrial carnivore definitive hosts and 7.09 % (95 % CI: 5.79-8.51) of intermediate hosts are infected. Among definitive hosts, opossums and foxes were most commonly infected with pooled global prevalence of 48.16 % (95 % CI: 14.62 - 82.69) and 35.97 % (295 % CI: 9.54 - 42.66) respectively. Pooled global prevalence in domestic dogs and cats were 7.97 % (95 % CI: 5.67 - 10.63) and 8.32 % (95 % CI: 3.78 - 14.41) respectively. Among intermediate hosts, birds and snakes were most commonly infected with pooled global prevalence of 16.19 % (95 %CI: 5.9 - 30.31) and 15.74 % (95 % CI: 10.59 - 21.69) respectively. Our analysis demonstrates that prevalence of Mesocestoides spp. is variable across the world. The sylvatic cycle in wild hosts is likely to be more important than the domestic cycle for the maintenance of Mesocestoides spp. globally. Currently available genetic data at the mitochondrial COI locus was also phylogenetically analyzed. The genetic data supports the taxonomic distinctiveness of only a few of the numerous morphologically described Mesocestoides spp.

First description of peritoneal and pleural metacestodosis caused by Mesocestoides vogae in a European wild cat (Felis silvestris silvestris).

Tapeworms of the genus Mesocestoides (Cestoda: Cyclophyllidea: Mesocestoididae) are still enigmatic to scientists, due to their high morphological variability, low host specificity, and unknown details of their life cycle. They are found worldwide, with carnivorous mammals as the main definitive hosts, and the disease is potentially zoonotic. After ingestion by a definitive host, the tetrathyridium can occasionally migrate through the intestinal wall and reach the peritoneal cavity or abdominal organs causing peritoneal metacestodosis. Here, we report on a case of metacestodosis of a European wild cat (Felis silvestris silvestris) found dead in Croatia. At necropsy, a large number of white, rice-like structures were found free in the abdominal and thoracic cavities, as well as along the serous surfaces and in the lungs. DNA isolated from the nodules was genotyped and based on a 320-base pair long 12S fragment classified as Mesocestoides vogae. Although post-mortem changes were advanced, severe emaciation due to the severe parasitic infection and gastrointestinal bleeding was diagnosed as the likely cause of death. Intestinal cestodosis was previously reported in wild cats, but according to our knowledge, this is the first description of peritoneal and pleural metacestodosis caused by M. vogae tetrathyridia (metacestodes) in any wild carnivore species.

Severe Pleural Effusion in a Dog Affected by Larval Mesocestodiasis.

A 4-year-old, 20 kg, female mixed breed dog was referred to our Veterinary Teaching Hospital for severe respiratory distress. Thoracic ultrasonography revealed severe pleural effusion with multiple anechoic cystic structures within the fluid. Abdominal cavity presented few small and irregular cystic structures. Diagnosis of infection by larval stages of Mesocestoides spp. on the basis of a PCR approach was performed and therapy with oral fenbendazole was started. Due to an incomplete clinical response, the dog underwent to remove metacestodes cysts by surgical debridement. The dog showed no recurrence of clinical signs for 12 months after the surgery. Pleural larval cestodiasis should be added as a differential diagnosis in the list of diseases causing pleural effusion in dogs.

Rodents as intermediate hosts of cestode parasites of mammalian carnivores and birds of prey in Poland, with the first data on the life-cycle of Mesocestoides melesi.

BACKGROUND: Rodents constitute an important part of the diet of many carnivore species. This predator-prey food chain is exploited by helminth parasites, such as cestodes, whose larval stages develop in rodents and then mature to the adult stage in predators. The main aim of our study was to use molecular techniques for identification of cestode species recovered from both intermediate and definitive hosts, with a particular focus on the genus Mesocestoides. METHODS: Larval cestodes were obtained during our long-term studies on rodent helminth communities in the Mazury Lake District in the north-east Poland in 2000-2018. Cestode larvae/cysts were collected from body cavities or internal organs (e.g. liver) during autopsies. Adult tapeworms were derived from nine red foxes, three Eurasian badgers and one Eurasian lynx. PCR amplification, sequencing and phylogenetic analyses were conducted employing three genetic markers: 18S rDNA, mitochondrial (mt) 12S rDNA and the mt cytochrome c oxydase subunit 1 (cox1) gene fragment. RESULTS: Altogether 19 Mesocestoides samples were analyzed, including 13 adult tapeworms from definitive hosts and six larval samples from 4 bank voles and 2 yellow-necked mice. Phylogenetic analyses revealed three well-supported trees of similar topology. In each case the Mesocestoides samples formed two separate clades. All isolates from foxes, the lynx isolate and two isolates from rodents grouped with Mesocestoides litteratus. Four isolates from rodents and all three isolates from Eurasian badgers were resolved in a separate clade, most similar to North American M. vogae (syn. M. corti). Examination of fixed, stained adult specimens from Eurasian badgers revealed consistency with the morphology of Mesocestoides melesi. Therefore, this clade is likely to represent M. melesi, a species first described in 1985 from the Eurasian badger Meles meles. Molecular analysis allowed also the identification of Taenia crassiceps, Hydatigera kamiyai and Cladotaenia globifera among larvae derived from rodents. CONCLUSIONS: Molecular and phylogenetic analyses support the recognition of M. melesi as a valid species. Our data represent the first record of the larvae of this species in rodents. This is the first report on the occurrence of H. kamiyai in rodents from Poland.

First molecular identification of the tapeworm Mesocestoides litteratus from an Italian wall lizard (Podarcis siculus).

The cyclophyllidean cestodes of the genus Mesocestoides are parasites infecting a variety of carnivorous vertebrates. The complete life cycle of these tapeworms is not clear yet, but some reptiles, amphibians, birds and micro-mammals have been indicated as intermediate hosts. Although Mesocestoides spp. are considered endemic in Europe, infections by the metacestode form, the tetrathyridium, are reported less commonly than are infections by adult worms. In the present study, a peritoneal infection by Mesocestoides tetrathyridia in a wall lizard (Podarcis siculus) collected in central Italy is described. The phylogenetic position of the metacestodes was assessed using the barcode region COI and species delimitation methods, i.e. Automatic Barcode Gap Discovery (ABGD) and Bayesian implementation of the Poisson Tree Processes model (bPTP). COI sequence analysis assigned the isolates from the wall lizard to Mesocestoides litteratus, as confirmed by species delimitation and phylogenetic analyses. M. litteratus has been observed in Europe mainly in the definitive hosts (dogs, cats and foxes), and never identified in the Mediterranean region. This study represents the first evidence of parasitic infection by tetrathyridia of M. litteratus in P. siculus in Italy.

Massive infection of a song thrush by Mesocestoides sp. (Cestoda) tetrathyridia that genetically match acephalic metacestodes causing lethal peritoneal larval cestodiasis in domesticated mammals.

BACKGROUND: Peritoneal larval cestodiasis induced by Mesocestoides Vaillant, 1863 (Cyclophyllidea: Mesocestoididae) is a common cause of severe infections in domestic dogs and cats, reported also from other mammals and less frequently from birds. However, there is a limited knowledge on the taxonomy of causative agents of this disease. RESULTS: In the present study, we investigated a massive, likely lethal, infection of a song thrush Turdus philomelos (Passeriformes: Turdidae) by Mesocestoides sp. tetrathyridia. We performed combined morphological and phylogenetic analysis of the tetrathyridia and compared them with the materials obtained previously from other birds and mammals. The metrical data fitted within the wide range reported by previous authors but confirmed the limited value of morphological data for species identification of tetrathyridia of Mesocestoides spp. The molecular analyses suggested that the isolates represented an unidentified Mesocestoides sp. that was previously repeatedly isolated and sequenced in larval and adult forms from domestic dogs and cats in Europe, the Middle East and North Africa. In contrast to the present study, which found encysted tetrathyridia, four of the five previous studies that identified the same species described infections by acephalic metacestodes only. CONCLUSIONS: The tetrathyridia of the examined Mesocestoides sp. are described in the present study for the first time. However, the possible match with the species that were previously reported to infect birds remains uncertain. The phylogenetic analyses also suggested the rejection of two cases that were previously identified as Mesocestoides corti as they were likely caused by the same species as in the presently reported infection case. The newly provided DNA sequences should allow the assignment to species in the future, when adults of the genus Mesocestoides are more thoroughly sequenced.

Transcriptomic profile of two developmental stages of the cestode parasite Mesocestoides corti.

Cestode development involves complex morphological and physiological changes. Here, we performed a differential expression analysis of gene transcripts between two developmental stages of the model cestode Mesocestoides corti. A RNA-seq-based approach was used to compare the transcriptomes of the tetrathyridium (larval, TT) and strobilated worm (ST) stages of the parasite. We found 19,053 transcripts, from which ∼45% were complete matches to genes previously annotated in the available M. corti draft genome sequence, ∼24% were considered novel isoforms, and ∼24% were considered potential novel transcripts. Stage-specific transcripts were found for both TTs (66) and STs (136), along with shared transcripts significantly overrepresented in one stage (342 in TTs, and 559 in STs). Differential expression and Gene Ontology term enrichment analyzes provided evidence of upregulation of different sets of transcripts associated with 'cytoskeleton', 'metabolism' and 'oxidation-reduction' processes in each stage, suggesting functional involvement of the corresponding genes with stage-specific features. Transcripts and processes enriched in the TT reflect typical larval processes that occur with the parasite in the intermediate host, such as asexual reproduction and budding, as well as active migration from the peritoneum to the liver and vice versa. In STs, transcripts associated with 'development', 'cell growth', and 'morphogenesis' were enriched, along with processes related to sexual reproduction, represented by the upregulation of numerous transcription factors, protein kinases, and histones. Overall, our results contributed to significantly increase the knowledge on the M. corti gene repertoire and expression profile in two developmental stages. Functional implications for the biology of larval and adult cestode parasites and for host-parasite interactions are discussed.

Species delimitation based on mtDNA genes suggests the occurrence of new species of Mesocestoides in the Mediterranean region.

BACKGROUND: This study is the first contribution to the molecular taxonomy of Mesocestoides spp. from domestic and wild carnivores in the Mediterranean area. A total of 13 adult worms and 13 larval stages of Mesocestoides spp. were collected from domestic and wild carnivore hosts in Italy and Tunisia. Samples collected in the Slovak Republic were used as comparative samples from outside the Mediterranean. The genes cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) of the mitochondrial genome were used as molecular markers to investigate the presence of cryptic Mesocestoides species in the area analysed. RESULTS: Results were consistent in showing three well-supported clusters of Mesocestoides spp. in southern Italy and Tunisia, which were strongly divergent from Mesocestoides litteratus, M. corti and M. lineatus. High levels of genetic variation and no evidence of geographical structuring was found between the clusters. CONCLUSIONS: Considering the low dispersal capability of the intermediate hosts of Mesocestoides spp., the lack of geographical structuring among the Mediterranean regions could be due to a high potential for dispersion of the definitive hosts. This study provides a foundation for future formal descriptions of new species of the genus Mesocestoides in the Mediterranean area.

Molecular identification of Mesocestoides sp. metacestodes in a captive gold-handed tamarin (Saguinus midas).

Mesocestoides spp. are tapeworms harbored in the intestine of many domestic and wild carnivores, birds of prey and, rarely, of humans. The life cycle is assumed to involve three hosts and the metacestode juvenile stage (tetrathyridium) may reproduce asexually, provoking severe systemic infections. In the present study, a case of a peritoneal infection by Mesocestoides sp. occurred in a captive gold-handed tamarin (Saguinus midas) is described. Phylogenetic positions based on CO1 and 12S loci were analysed to describe in details this unusual finding of Mesocestoides in a callitrichid monkey. The phylogenetic analyses has not clearly resolve relationships between the metacestode forms from the tamarin and any of the described Mesocestoides species available so far, including those described from non-human primates, thus supporting the hypothesis of a new taxon within the genus.

Gastrointestinal helminths of gray wolves (Canis lupus lupus) from Sweden.

As the Scandinavian wolf population is limited in size, it is only rarely subject to systematic studies on its disease biology, especially gastrointestinal parasites. Therefore, this study aims to describe the prevalence of gastrointestinal helminths of gray wolves hunted on a limited license as a part of a wildlife management program. Helminths of 20 wolves were examined post mortem by macroscopy and coprology. Intestinal worms of five species were recovered from 18 wolves (90%): Uncinaria stenocephala (90%), Taenia spp. (45%), Alaria alata (25%), and Mesocestoides spp. (5%). Of the taeniid specimens typed by multiplex PCR and sequencing of the cox1 gene, 25% belonged to Taenia hydatigena and 25% to Taenia krabbei. The overall species diversity was low compared to findings from wolves of the northern hemisphere. Fecal eggs of Eucoleus boehmi were detected in 12 wolves (60%). Fecal metastrongylid larvae were found in seven individuals (39%), but PCR analyses specific for Angiostrongylus vasorum were negative. The wolves were in good body condition suggesting that the parasite infestation had no negative impact on the general health of the examined wolves. Although some of the recovered parasite species have zoonotic or veterinary impact, it is not likely that the spare wolf population pose substantial threat to human or veterinary health.

Mesocestoides sp. in Wild Northern Bobwhite ( Colinus virginianus) and Scaled Quail ( Callipepla squamata).

Mesocestoides spp. have a cosmopolitan distribution with zoonotic potential. Mesocestoides tetrathyridia were found under the pericardial sac, on the surface of the crop, and in the peritoneal cavity of a hunter-harvested Northern Bobwhite ( Colinus virginianus) and a Scaled Quail ( Callipepla squamata) collected during the 2016-17 quail hunting season in northwest and southern Texas, US respectively. Molecular analysis indicated that the tetrathyridia from the birds likely belonged to an undescribed species and are identical to pretetrathyridium stages recently found in Scincella lateralis skinks in Oklahoma. This is the first report of Mesocestoides from North American birds.

Unique pharmacological properties of serotoninergic G-protein coupled receptors from cestodes.

BACKGROUND: Cestodes are a diverse group of parasites, some of them being agents of neglected diseases. In cestodes, little is known about the functional properties of G protein coupled receptors (GPCRs) which have proved to be highly druggable targets in other organisms. Notably, serotoninergic G-protein coupled receptors (5-HT GPCRs) play major roles in key functions like movement, development and reproduction in parasites. METHODOLOGY/PRINCIPAL FINDINGS: Three 5-HT GPCRs from Echinococcus granulosus and Mesocestoides corti were cloned, sequenced, bioinformatically analyzed and functionally characterized. Multiple sequence alignment with other GPCRs showed the presence of seven transmembrane segments and conserved motifs but interesting differences were also observed. Phylogenetic analysis grouped these new sequences within the 5-HT7 clade of GPCRs. Molecular modeling showed a striking resemblance in the spatial localization of key residues with their mammalian counterparts. Expression analysis using available RNAseq data showed that both E. granulosus sequences are expressed in larval and adult stages. Localization studies performed in E. granulosus larvae with a fluorescent probe produced a punctiform pattern concentrated in suckers. E. granulosus and M. corti larvae showed an increase in motility in response to serotonin. Heterologous expression revealed elevated levels of cAMP production in response to 5-HT and two of the GPCRs showed extremely high sensitivity to 5-HT (picomolar range). While each of these GPCRs was activated by 5-HT, they exhibit distinct pharmacological properties (5-HT sensitivity, differential responsiveness to ligands). CONCLUSIONS/SIGNIFICANCE: These data provide the first functional report of GPCRs in parasitic cestodes. The serotoninergic GPCRs characterized here may represent novel druggable targets for antiparasitic intervention.

Morphological and Molecular Characterization of Post-Larval Pre-Tetrathyridia of Mesocestoides sp. (Cestoda: Cyclophyllidea) from Ground Skink, Scincella lateralis (SAURIA: SCINCIDAE), FROM SOUTHEASTERN OKLAHOMA.

Free pre-tetrathyridia of Mesocestoides sp. are described, for the first time, from samples obtained from the coelomic cavity of a ground skink, Scincella lateralis, from Oklahoma. Closer examination of these early-stage tapeworms revealed they were transitional metamorphosis stages between a post-hexacanth procercoid form to the full metacestode of Mesocestoides. A series of transitional stages was found that span the full period of sucker and apical organ development. However, we did not see any fully developed tetrathyridia, i.e., having classic Mesocestoides morphology but with the apical sucker absent following developmental atrophy. This is the first time that metamorphic pre-tetrathyridial stages of a Mesocestoides sp. have been reported in vivo from a natural infection. These observations corroborate earlier reports of such stages of Mesocestoides vogae developed in vitro, though the previously reported isolate of M. vogae is asexually proliferative, and the species from the present study showed no sign of asexual proliferation. The fact that these immediately post-hexacanth stages can occur in a single lizard intermediate host may suggest that Mesocestoides spp. might develop through a simple 2-host life cycle rather than an obligate 3-host cycle that has been speculated to occur by most previous authors. DNA sequence comparisons and phylogenetic analyses based on mitochondrial 12S, cox1, and nad1 genes have demonstrated that our specimens from S. lateralis represent a species clearly distinct from all previously sequenced Mesocestoides and closely related to 2 forms from domestic dogs and Channel Island fox in California previously published as Mesocestoides sp. C.

Small rodents as paratenic or intermediate hosts of carnivore parasites in Berlin, Germany.

Rodents are important intermediate and paratenic hosts for carnivore parasites, including the important zoonotic agents Toxoplasma, Echinococcus and Toxocara. Monitoring of such parasites in rodents can be used to detect increasing risks for human and veterinary public health. Rodents were trapped at four sites in Berlin, two near the city center, two at the periphery. PCRs were conducted to detect Coccidia (target ITS-1) and specifically Toxoplasma gondii (repetitive element) in brain and ascarids (ITS-2) in muscle or brain tissue. During necropsies, metacestodes were collected and identified using ITS-2 and 12S rRNA PCRs. An ELISA to detect antibodies against Toxocara canis ES antigens was performed. Within the 257 examined rodents, the most frequently observed parasite was Frenkelia glareoli predominantly found in Myodes glareolus. T. gondii was only detected in 12 rodents and Microtus spp. (although strongly underrepresented) had a significantly increased chance of being positive. Neither Echinococcus nor typical Taenia parasites of dogs and cats were found but Mesocestoides litteratus and Taenia martis metacestodes were identified which can cause severe peritoneal or ocular cysticercosis in dogs, primates and humans. Using PCR, the ascarids T. canis (n = 8), Toxocara cati (4) and Parascaris sp. (1) were detected predominantly in muscles. Seroprevalence of T. canis was 14.2% and ELISA was thus more sensitive than PCR to detect infection with this parasite. Non-parametric multidimensional scaling and cluster analysis revealed that parasite communities could be grouped into an urban and a peri-urban cluster with high frequency of ascarid-positive rodents in urban and high frequency of F. glareoli in peri-urban sites. Prevalence rates of parasites in rodents with potential impact for human or veterinary public health are considerable and the monitoring of transmission cycles of carnivore parasites in intermediate rodent hosts is recommended to estimate the health risks arising from wild and domesticated carnivores.

Proteomic Analysis of Excretory-Secretory Products of Mesocestoides corti Metacestodes Reveals Potential Suppressors of Dendritic Cell Functions.

Accumulating evidences have assigned a central role to parasite-derived proteins in immunomodulation. Here, we report on the proteomic identification and characterization of immunomodulatory excretory-secretory (ES) products from the metacestode larva (tetrathyridium) of the tapeworm Mesocestoides corti (syn. M. vogae). We demonstrate that ES products but not larval homogenates inhibit the stimuli-driven release of the pro-inflammatory, Th1-inducing cytokine IL-12p70 by murine bone marrow-derived dendritic cells (BMDCs). Within the ES fraction, we biochemically narrowed down the immunosuppressive activity to glycoproteins since active components were lipid-free, but sensitive to heat- and carbohydrate-treatment. Finally, using bioassay-guided chromatographic analyses assisted by comparative proteomics of active and inactive fractions of the ES products, we defined a comprehensive list of candidate proteins released by M. corti tetrathyridia as potential suppressors of DC functions. Our study provides a comprehensive library of somatic and ES products and highlight some candidate parasite factors that might drive the subversion of DC functions to facilitate the persistence of M. corti tetrathyridia in their hosts.

Identification and profiling of microRNAs in two developmental stages of the model cestode parasite Mesocestoides corti.

MicroRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in fundamental biological processes such as metabolism and development. The particular developmental characteristics of cestode parasites highlight the importance of studying miRNA gene regulation in these organisms. Here, we performed a comprehensive analysis of miRNAs in two developmental stages of the model cestode Mesocestoides corti. Using a high-throughput sequencing approach, we found transcriptional evidence of 42 miRNA loci in tetrathyridia larvae and strobilated worms. Tetrathyridium and strobilated worm-specific miRNAs were found, as well as differentialy expressed miRNAs between these developmental stages, suggesting miRNA regulation of stage-specific features. Moreover, it was shown that uridylation is a differential mechanism of post-transcriptional modification of M. corti miRNAs. The whole set of M. corti miRNAs represent 33 unique miRNA families, and confirm the remarkable loss of conserved miRNA families within platyhelminth parasites, reflecting their relatively low morphological complexity and high adaptation to parasitism. Overall, the presented results provide a valuable platform to studies aiming to identify and characterize novel miRNA-based molecular mechanisms of post-transcriptional gene regulation in cestodes, necessary for the elucidation of developmental aspects of the complex biology of these parasites.

Occurrence of Mesocestoides canislagopodis (Rudolphi, 1810) (Krabbe, 1865) in mammals and birds in Iceland and its molecular discrimination within the Mesocestoides species complex.

The life cycle of Mesocestoides tapeworms (Cestoda: Cyclophyllidea: Mesocestoididae) requires three hosts. The first intermediate host is unknown but believed to be an arthropod. The second intermediate host is a vertebrate. The primary definitive host is a carnivore mammal, or a bird of prey, that eats the tetrathyridium-infected second intermediate host. One representative of the genus, Mesocestoides canislagopodis, has been reported from Iceland. It is common in the arctic fox (Vulpes lagopus) and has also been detected in domestic dogs (Canis familiaris) and cats (Felis domestica). Recently, scolices of a non-maturing Mesocestoides sp. have also been detected in gyrfalcon (Falco rusticolus) intestines, and tetrathyridia in the body cavity of rock ptarmigan (Lagopus muta). We examined the taxonomic relationship of Mesocestoides from arctic fox, gyrfalcon, and rock ptarmigan using molecular methods, both at the generic level (D1 domain LSU ribosomal DNA) and at the specific level (cytochrome c oxidase subunit I (COI) and 12S mitochondrial DNA). All stages belonged to Mesocestoides canislagopodis. Phylogenetic analysis of the combined 12S-COI at the specific level confirmed that M. canislagopodis forms a distinct clade, well separated from three other recognized representatives of the genus, M. litteratus, M. lineatus, and M. corti/vogae. This is the first molecular description of this species. The rock ptarmigan is a new second intermediate host record, and the gyrfalcon a new primary definitive host record. However, the adult stage seemed not to be able to mature in the gyrfalcon, and successful development is probably restricted to mammalian hosts.