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The cestode Echinococcus multilocularis is the causative agent of alveolar echinococcosis, a fatal zoonotic parasitic disease of the northern hemisphere. Red foxes are the main reservoir hosts and, likely, the main drivers of the geographic spread of the disease in Europe. Knowledge of genetic relationships among E. multilocularis isolates at a European scale is key to understanding the dispersal characteristics of E. multilocularis. Hence, the present study aimed to describe the genetic diversity of E. multilocularis isolates obtained from different host species in 19 European countries. Based on the analysis of complete nucleotide sequences of the cob, atp6, nad2, nad1 and cox1 mitochondrial genes (4,968 bp), 43 haplotypes were inferred. Four haplotypes represented 62.56 % of the examined isolates (142/227), and one of these four haplotypes was found in each country investigated, except Svalbard, Norway. While the haplotypes from Svalbard were markedly different from all the others, mainland Europe appeared to be dominated by two main clusters, represented by most western, central and eastern European countries, and the Baltic countries and northeastern Poland, respectively. Moreover, one Asian-like haplotype was identified in Latvia and northeastern Poland. To better elucidate the presence of Asian genetic variants of E. multilocularis in Europe, and to obtain a more comprehensive Europe-wide coverage, further studies, including samples from endemic regions not investigated in the present study, especially some eastern European countries, are needed. Further, the present work proposes historical causes that may have contributed to shaping the current genetic variability of E. multilocularis in Europe., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Human alveolar echinococcosis (AE) is a serious parasitic disease caused by larval stages of Echinococcus multilocularis. Between January 2000 and October 2023, 137 AE cases were confirmed in Slovakia. The average annual incidence increased from 0.031 per 100,000 inhabitants between 2000 and 2011, to an average of 0.187 since 2012, i.e. about six times. Among patients, 45.3% were men and 54.7% were women; the mean age at the time of diagnosis was 52.8 years. Most cases were diagnosed in the age groups 51-60 years and 61-70 years (33 cases each), and eight patients fell into the age category ≤ 20 years. To better recognize the gene diversity in clinical samples, metacestodes from 21 patients collected between 2013 and 2021 were subjected to DNA sequencing of four mitochondrial genes. Using concatenated sequences of cob (603 bp), nad2 (882 bp) and cox1 (789 bp) gene fragments, 14 isolates (66.7%) were assigned to the European E5 profile of E. multilocularis, two isolates (9.5%) to the E5a subtype, four isolates (19%) to the E4 profile, and one isolate (4.8%) to haplogroup E1/E2. The E5-type profiles and E4 profiles were distributed throughout the country, whereas the E1/E2 profile was found in the patient from western Slovakia. According to the data obtained and GenBank sequences, the E5-type dispersal is so far limited to central-eastern Europe and the variant seems to be indigenous to that region. The admixture with the haplotypes E4 and E1/E2 could have taken place from a historical endemic focus during the fox expansion in the last decades. By employing the nad1 fragment, a typical European haplotype was observed in all 21 resolved Slovak samples. The acceleration in the AE incidence in the last decade suggests the emergence of the disease and the need for further research on human and animal isolates., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Antolová et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Alveolar echinococcosis, caused by the larval stages of the tapeworm Echinococcus multilocularis (Leuckart, 1863), is of increasing concern in the northern hemisphere. Most cases of alveolar echinococcosis (excluding Alaska) appear to be linked with European and Asian genotypes that highlight the need for a more precise delimiting of their actual distribution and tracing historical episodes of their translocations and introductions into new areas. We have herein summarized previous available research studies, which mentioned firstly described geographic M1/M2 variants of E. multilocularis using molecular tools (established by sequencing of mitochondrial genes cox1, 366 bp and nad1, 471 bp), in an attempt to consolidate their correct affiliations with the geographic origin in sense of the original description from the early 1990´s. Since 2009, inverted designations (M1 named as M2 and vice versa) are being prevailing in research literature (we found ten erroneous vs. three correct classifications) that might bias genetic interpretation of comparative data in specific cases. When comparing M1/M2 profiles to those obtained from mitochondrial evidences over the last decades, the phylogenetic analysis revealed that the M1 strain (originally described from China, Alaska, North America) grouped with the Asian clade of E. multilocularis more recently established, whereas the M2 strain (described from the German vole) had a specific structure, in cox1 clustering with the North American clade. It is presumed that events of intercontinental expansion and isolation covering glacial and interglacial periods during the late Pleistocene have likely accounted for the transmission of this discrete genotype from Beringia into endemic area of western and central Europe via circumpolar movements of foxes., Competing Interests: Declaration of Competing Interest The authors state that they have no conflict of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)