Unique genetic structure of the human tapeworm Dibothriocephalus latus from the Alpine lakes region - a successful adaptation?

Dibothriocephalus latus is the most frequent causative agent of fish-borne zoonosis (diphyllobothriosis) in Europe, where it is currently circulating mainly in the Alpine lakes region (ALR) and Russia. Three mitochondrial genes (cox1, cob and nad3) and 6 microsatellite loci were analysed to determine how is the recently detected triploidy/parthenogenesis in tapeworms from ALR displayed at the DNA level. A geographically distant population from the Krasnoyarsk Reservoir in Russia (RU-KR) was analysed as a comparative population. One or 2 alleles of each microsatellite locus was detected in plerocercoids from RU-KR, corresponding to the microsatellite pattern of a diploid organism. In contrast, 1­3 alleles were observed in tapeworms from ALR, in accordance with their triploidy. The high diversity of mitochondrial haplotypes in D. latus from RU-KR implied an original and relatively stable population, but the identical structure of mitochondrial genes of tapeworms from ALR was probably a consequence of a bottleneck typical of introduced populations. These results indicated that the diploid/sexually reproducing population from RU-KR was ancestral, located within the centre of the distribution of the species, and the triploid/parthenogenetically reproducing subalpine population was at the margin of the distribution. The current study revealed the allelic structure of the microsatellite loci in the triploid tapeworm for the first time.

Mitochondrial and microsatellite data show close genetic relationships between Dibothriocephalus latus from South America (Argentina) and Europe (the Alpine lakes region).

The most frequent causative agent of diphyllobothriosis, a fish-borne parasitic zoonosis, is the broad fish tapeworm Dibothriocephalus latus distributed mainly throughout the Holarctic region. The larval stages of the tapeworm were also detected in native and introduced freshwater fish in several lakes in South America, particularly in the north-western Patagonia in Argentina. The main objective of the present study was to determine the genetic structure of D. latus from rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta), and brook trout (Salvelinus fontinalis) from Gutiérrez Lake and Alicura Reservoir in Argentina using the sequences of three mitochondrial genes (cox1, cob, and nad3) and six polymorphic microsatellite loci. The results were compared with the corresponding molecular data of D. latus from Europe (Switzerland and Italy; the Alpine lakes region) and Asia (Siberia, Russia). Only one concatenated haplotype identical with the con-Ha1 specific for D. latus from the Alpine lakes region was detected in all individuals from Argentina. Three different alleles were detected in five out of six analysed microsatellite loci, indicating the presence of three sets of chromosomes. The same structure of microsatellite loci was recently observed in D. latus from Switzerland and Italy, in which triploidy was previously confirmed by cytogenetic study. The data on the mitochondrial genes, the allelic structure of microsatellite loci, and the principal coordinate analysis showed close genetic relationships between D. latus from Argentina and the Alpine lakes region, thus supporting the hypothesis of the European origin of the broad fish tapeworm from South America.

Molecular cytogenetic analysis of a triploid population of the human broad tapeworm, Dibothriocephalus latus (Diphyllobothriidea).

The large-sized tapeworm Dibothriocephalus latus is known as the broad or fish-borne cestode of mammals that is capable to infect humans and cause diphyllobothriosis. Recently, molecular data on D. latus has been accumulating in the literature and a complete genome sequence has been published; however, little is known about the karyotype and chromosome architecture. In this study, an in-depth karyological analysis of 2 D. latus specimens was carried out. The plerocercoids originated from a perch caught in subalpine Lake Iseo (Italy) and the tapeworms were reared in hamsters. Both specimens contained cells with a highly variable number of chromosomes ranging from18 to 27. Nevertheless, the largest portion of mitotic figures (47%) showed a number corresponding to the triploid set, 3n = 27. Accordingly, the karyotype of the analyzed specimens consisted of 9 triplets of metacentric chromosomes. Fluorescence in situ hybridization (FISH) with the 18S rDNA probe clearly demonstrated the presence of 3 clusters of hybridization signals on the triplet of chromosome 7, thus confirming the triploid status of the specimens. FISH with a telomeric (TTAGGG)n probe confined hybridization signals exclusively to the terminal chromosomal regions, supporting the earlier findings that this repetitive motif is a conserved feature of tapeworm telomeres.

The first records of Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidae), a causative agent of human sparganosis, in Latvian wildlife.

Diphyllobothriid tapeworms of the genus Spirometra are causative agents of sparganosis, food-borne zoonotic parasitic disease. They have been recorded in broad spectrum of hosts, including humans, in all continents except Antarctica. Spirometra tapeworms have been intensively studied in several Asian countries; however, they have been rather neglected in Europe. The aim of this study was to provide a pilot screening of Spirometra spp. in Latvia, where data on sparganosis are not available. Tapeworms morphologically identified as diphyllobothriid species were isolated from grey wolves Canis lupus and Eurasian lynxes Lynx lynx from Latvia during the hunting periods 2013-2019. The parasites were subjected to molecular genotyping using sequences of the partial large (LSU rDNA; 615 bp) and small (SSU rDNA; 720 bp) subunits of the nuclear ribosomal RNA gene and complete (1566 bp) cytochrome c oxidase subunit I gene of the mitochondrial DNA (cox1 mtDNA). Analyses of both ribosomal subunits of 13 tapeworms revealed no intraspecific variation within the respective rDNA subunits. On the other hand, sequence analysis of mitochondrial cox1 revealed intraspecific polymorphism displayed by 12 cox1 haplotypes. Comparison of the current data with sequences of the corresponding DNA regions deposited in the GenBank revealed 99.3-99.5% (LSU rDNA), 99.2% (SSU rDNA) and 99.6-100% (cox1 mtDNA) identity of studied tapeworms with Spirometra erinaceieuropaei, which provided the first confirmation of this diphyllobothriid tapeworm in Latvia. Since S. erinaceieuropaei is probably prevalent in Latvian wildlife and may also occur in other potential host species, further studies are needed in order to acquire complex data on its geographic distribution and transmission in the natural environment of Latvia, as well as on the spectrum of its intermediate, paratenic, and definitive hosts.

Host Switching of Zoonotic Broad Fish Tapeworm (Dibothriocephalus latus) to Salmonids, Patagonia.

Diphyllobothriosis is a reemerging zoonotic disease because of global trade and increased popularity of eating raw fish. We present molecular evidence of host switching of a human-infecting broad fish tapeworm, Dibothriocephalus latus, and use of salmonids as intermediate or paratenic hosts and thus a source of human infection in South America.

Ups and downs of infections with the broad fish tapeworm Dibothriocephalus latus in Europe (Part II) and Asia from 1900 to 2020.

The broad fish tapeworm, Dibothriocephalus latus (Diphyllobothriidea), is the most important causative agent of diphyllobothriosis, a fish-borne zoonosis, in Europe. Part I of this review focused on the occurrence of D. latus in northwestern and central Europe, particularly in Fennoscandia, the Baltic, the Alpine lakes and Danube River regions during 1900-2020. Part II summarises data on D. latus from the European and Asian parts of Russia and from Asian countries. The tapeworm has occurred throughout Russia, with the most important foci in (i) the Republic of Karelia in the northwest of European Russia, (ii) the Volga River basin in the central and southern parts of European Russia, (iii) the Ob-Irtysh rivers region in the Ural region, (iv) the Yenisei-Lena rivers region in Siberia, and (v) the Lake Baikal basin in Siberia. The incidence of diphyllobothriosis has declined in recent decades, especially in European Russia, but zoonosis is still prevalent in some regions of Siberia. Cases reported from Arctic regions, the region around Lake Baikal, and the Pacific coast, including the Amur basin, however, were probably misidentifications with D. dendriticus and/or D. nihonkaiensis. No other Asian country where D. latus findings represented either imported cases or misidentifications had natural focus of diphyllobothriosis. Patterns of distribution of D. latus occurrence were similar in all Eurasian foci between 1900 and 2020. The numbers of records were associated with historical and epidemiological milestones of particular time periods.

Bird Louse Flies Ornithomya spp. (Diptera: Hippoboscidae) as Potential Vectors of Mammalian Babesia and Other Pathogens.

Background: Birds and mammals share various ectoparasites, which are responsible for the transmission of a wide range of pathogens. The louse flies (family Hippoboscidae) are ectoparasitic dipterans feeding strictly on the blood of mammals and birds. Both sexes of the louse flies are obligatory hematophagous and are known to act as the vectors of infectious agents. Materials and Methods: A total of 20 specimens of Ornithomya sp. were collected by hand on birds caught in nets or by hand from humans in two localities in Eastern Slovakia in 2021. The DNA samples were individually screened by species-specific PCRs for the presence of selected vector-borne pathogens. Results: Taxonomic identification folowed by molecular analyses revealed two louse fly species of Ornithomya spp. (O. avicularia and O. biloba). The molecular screening provided negative PCR results for Anaplasma phagocytophilum, Borrelia burgdorferi s.l., Rickettsia spp., Bartonella spp., and Hepatozoon canis. In contrast, positive PCR results were obtained for Babesia spp., Wolbachia spp., and Trypanosoma corvi. Conclusions: Of epidemiological importance is that the louse flies can presumably spread Babesia and other pathogens by host switching which facilitates the transmission and spread of numerous pathogens.

Genetic interrelationships of Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidea), the causative agent of sparganosis in Europe.

The geographic distribution of Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidea), the causative agent of food/water-borne sparganosis, is restricted to Europe, where infected canids, felids, mustelids, suids, and reptiles have been documented from Poland, Ukraine, Belarus, Russia, Serbia, Estonia, Latvia, and Finland. The main objective of the current study was to map the molecular divergence of S. erinaceieuropaei from Finland using the complete sequences of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1 mtDNA). Seven cox1 haplotypes were determined in 15 tapeworms from Eurasian lynx (Lynx lynx) from three localities in southern Finland. In addition, the first inter-population study of S. erinaceieuropaei based on currently obtained data on cox1 from Finland and previously published data from Finland, Latvia, Ukraine, and Poland, was performed. The haplotype network showed a star-like pattern without specific subdivision of lineages according to the locality. Samples from Finland, Latvia, and Poland shared several haplotypes and formed the common Baltic lineage. The haplotype of S. erinaceieuropaei from Ukraine was unique and placed on a separate mutational pathway, suggesting a different lineage of the parasite.

TITLE: Interrelations génétiques de Spirometra erinaceieuropaei (Cestoda, Diphyllobothriidea), l'agent causal de la sparganose en Europe. ABSTRACT: La distribution géographique de Spirometra erinaceieuropaei (Cestoda : Diphyllobothriidea), l'agent causal de la sparganose d'origine alimentaire/hydrique, est limitée à l'Europe, où des canidés, félidés, mustélidés, suidés et reptiles infectés ont été documentés en Pologne, Ukraine, Biélorussie, Russie, Serbie, Estonie, Lettonie et Finlande. L'objectif principal de la présente étude était de cartographier la divergence moléculaire de S. erinaceieuropaei de Finlande à l'aide des séquences complètes du gène mitochondrial de la sous-unité 1 de la cytochrome c oxydase (ADNmt cox1). Sept haplotypes cox1 ont été déterminés chez quinze cestodes du Lynx d'Eurasie (Lynx lynx) de trois localités du sud de la Finlande. En outre, la première étude inter-populationnelle de S. erinaceieuropaei basée sur les données actuellement obtenues sur cox1 de Finlande et sur des données précédemment publiées de Finlande, Lettonie, Ukraine et Pologne, a été réalisée. Le réseau d'haplotypes a montré un motif en étoile sans subdivision spécifique des lignées selon la localité. Des échantillons de Finlande, Lettonie et Pologne partagent plusieurs haplotypes et forment la lignée commune de la Baltique. L'haplotype de S. erinaceieuropaei d'Ukraine est unique et placé sur une voie de mutation distincte suggérant une lignée différente du parasite.

Ups and downs of infections with the broad fish tapeworm Dibothriocephalus latus in Europe from 1900 to 2020: Part I.

The broad fish tapeworm, Dibothriocephalus latus (Diphyllobothriidea), is the most frequent causative agent of diphyllobothriosis, a fish-borne zoonosis, in Europe. Diphyllobothriosis is characterized by the transmission of D. latus larvae to humans via the consumption of raw, marinated, smoked or inadequately cooked fish products. The most important European foci of diphyllobothriosis have been Fennoscandia, the Baltic region, the Alpine lakes region, the Danube River region, and several endemic regions in Russia. This review provides basic data on the biology, life cycle, host specificity, methods of identification of D. latus, and a detailed summary of its occurrence in intermediate and definitive hosts in Fennoscandia and the Baltic, Alpine, and Danube regions during the last 120 years (1900-2020). Deeper insight into the unique pattern of distribution of D. latus in endemic regions is provided. The numbers of records are associated with several milestones of particular time periods. The first milestone (historical), which influenced studies on D. latus in Europe, was the period during and after World War II (1941-1950). The second milestone (epidemiological) was the decade 1981-1990, when previous massive health campaigns led to a marked decline of diphyllobothriosis in Europe and less published data on D. latus. Based on recent data, the broad fish tapeworm is either absent or present at very low prevalences in Fennoscandia and the Baltic and Danube regions, but the Alpine lakes region represents a continuous ongoing circulation of the parasite in the natural environment and humans.

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.

A study of the endohelminths of the European perch Perca fluviatilis L. from the central region of the Danube river basin in Slovakia.

The European perch Perca fluviatilis L. serves as a host of different endohelminths of Trematoda, Cestoda, Nematoda, and Acanthocephala. Its natural range covers freshwater basins throughout much of Europe, including the Danube. Since information about endohelminths of European perch from this international river basin has been rather sporadic, the parasitological examinations of 700 perch from the central region of the Danube river basin in Slovakia were performed in October 2017 and April 2018. The larval stages of Triaenophorus nodulosus (Cestoda) were found in cysts located in the perch liver and adults of Proteocephalus percae (Cestoda) were isolated from the intestine. The larval stages of Eustrongylides sp. (Nematoda) and metacercariae of Clinostomum complanatum (Trematoda), both potential causative agents of fish-borne zoonoses, were found in the musculature. Spatial and seasonal differences in the occurrence of currently detected helminths were discussed with data on biological and environmental conditions of particular sampling site.

Endohelminths of European Perch (Perca fluviatilis) from Selected Localities in Poland with an Emphasis on Search of the Broad Fish Tapeworm Dibothriocephalus latus.

INTRODUCTION: A piscivorous fish European perch (Perca fluviatilis) is present in all types of lakes and brackish waters in Poland. Previous ichthyoparasitological surveys revealed broad spectrum of endohelminths in perch from different aquatic environments. Among them, detection of Diphyllobothrium sp. and Dibothriocephalus latus (syn. Diphyllobothrium latum) in the Pomeranian Bay is of particular interest, since D. latus is one of the causative agents of diphyllobothriasis, fish-borne parasitic zoonosis. Besides, D. latus eggs were previously detected in coprological samples of otter, wolf and lynx from the Bialowieza Primeval Forest. AIM: To conduct parasitological examinations of European perch from the Pomeranian Bay in order to detect the spectrum of its endohelminths and to provide a pilot study on helminths of perch from different water bodies in the Bialowieza Primeval Forest. Due to zoonotic character of D. latus, we have focused our attention to this tapeworm. RESULTS: The larvae of tapeworm Triaenophorus nodulosus and thorny-headed worm Acanthocephalus lucii were detected in perch from the Pomeranian Bay. In perch from different localities in the Bialowieza Primeval Forest, T. nodulosus, A. lucii and tapeworm Proteocephalus percae were detected. D. latus plerocercoids were found neither in musculature nor in peritoneal cavity and other internal organs of any of the fish examined from both studied localities in Poland. CONCLUSION: Future screening implementing morphological and molecular markers is needed in order to understand the current distribution of D. latus in Europe.