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This study aimed to fill a gap of knowledge by providing a quantitative measure of molecularly identified species and genotypes belonging to Echinococcus granulosus sensu lato (s.l.) causing human cystic echinococcosis (CE) in Europe during the period 2000-2021. As these species and genotypes are characterized by genetic, animal host and geographical differences, studying the E. granulosus s.l. complex is epidemiologically relevant. A systematic review (SR) was conducted on the basis of both scientific and grey literature considering primary studies between 2000 and 2021 in four databases. From a total of 1643 scientific papers, 51 records were included in the SR. The main inclusion criterion for this study was the molecular confirmation of E. granulosus s.l. at the genotype/species level as a causative agent of human CE cases in selected European countries. Relevant data were obtained from 29 out of 39 eligible European countries. This SR identified 599 human molecularly confirmed echinococcal cysts: 460 (76.8%) identified as E. granulosus sensu stricto (s.s.), 130 (21.7%) as E. canadensis cluster (G6/7 and G10), 7 (1.2%) as E. ortleppi (G5), and 2 as E. vogeli (0.3%). Three geographical hotspots of human CE caused by different species of the E. granulosus s.l. complex were identified: (1) E. granulosus s.s. in Southern and South-eastern Europe (European-Mediterranean and Balkan countries); (2) E. canadensis (G6/7) in Central and Eastern Europe; (3) E. ortleppi in Central and Western Europe. This SR also identified data gaps that prevented a better definition of the geographical distribution of the Echinococcus granulosus s.l. species complex in Europe: western Balkan countries, part of Central Europe, and Baltic countries. These results mandate longitudinal, multi-centre, intersectoral and transdisciplinary studies which consider both molecular and clinical epidemiology in animals and humans. Such studies would be valuable for a better understanding of the transmission of the E. granulosus s.l. species complex and their potential clinical impact on humans.

Shengyue Wang and colleagues report the draft genome sequence and transcriptome analysis for Echinococcus granulosus , a parasitic helminth and cause of human hydatid disease. Their comparative genomic analysis identifies genes acquired by E. granulosus that are associated with host immune response, parasite survival and growth. Cystic echinococcosis (hydatid disease), caused by the tapeworm E. granulosus , is responsible for considerable human morbidity and mortality. This cosmopolitan disease is difficult to diagnose, treat and control. We present a draft genomic sequence for the worm comprising 151.6 Mb encoding 11,325 genes. Comparisons with the genome sequences from other taxa show that E. granulosus has acquired a spectrum of genes, including the EgAgB family, whose products are secreted by the parasite to interact and redirect host immune responses. We also find that genes in bile salt pathways may control the bidirectional development of E. granulosus , and sequence differences in the calcium channel subunit EgCa v β 1 may be associated with praziquantel sensitivity. Our study offers insights into host interaction, nutrient acquisition, strobilization, reproduction, immune evasion and maturation in the parasite and provides a platform to facilitate the development of new, effective treatments and interventions for echinococcosis control.

Tapeworms (Cestoda) cause neglected diseases that can be fatal and are difficult to treat, owing to inefficient drugs. Here we present an analysis of tapeworm genome sequences using the human-infective species Echinococcus multilocularis , E. granulosus, Taenia solium and the laboratory model Hymenolepis microstoma as examples. The 115- to 141-megabase genomes offer insights into the evolution of parasitism. Synteny is maintained with distantly related blood flukes but we find extreme losses of genes and pathways that are ubiquitous in other animals, including 34 homeobox families and several determinants of stem cell fate. Tapeworms have specialized detoxification pathways, metabolism that is finely tuned to rely on nutrients scavenged from their hosts, and species-specific expansions of non-canonical heat shock proteins and families of known antigens. We identify new potential drug targets, including some on which existing pharmaceuticals may act. The genomes provide a rich resource to underpin the development of urgently needed treatments and control. Genome sequences of human-infective tapeworm species reveal extreme losses of genes and pathways that are ubiquitous in other animals, species-specific expansions of non-canonical heat shock proteins and families of known antigens, specialized detoxification pathways, and metabolism that relies on host nutrients; this information is used to identify new potential drug targets. Four tapeworm genomes sequenced Tapeworms cause echinococcosis and cysticercosis, two of the most severe parasitic diseases found in humans, and both on the World Health Organization's list of neglected tropical diseases. The publication of four tapeworm genome sequences — human-infective tapeworm species Echinococcus multilocularis , E. granulosus , Taenia solium and the laboratory model Hymenolepis microstoma — and identification of potential new drug targets for treating tapeworm infections is therefore a welcome development. Analysis of the sequences provides insights into the evolution of parasitism and reveals extreme losses of genes and pathways ubiquitous in other animals on one hand and species-specific expansions of genes on the other. More than a thousand E. multilocularis proteins emerge as potential targets, and of these, close to 200 with the highest scores may be targeted with existing pharmaceuticals.

The island of Cyprus was a historical endemic area for cystic echinococcosis (CE) in the Mediterranean. During the last decades, Cyprus has been an open-air laboratory and a model for testing and implementing control measures aiming to eliminate CE as a public health problem. Despite control and surveillance measures implemented during last 50 years, molecular characterization of Echinococcus granulosus sensu lato specimens has been never provided. In February 2023, the carcass of a stray dog collected in the Nicosia district was examined by the Veterinary Services and found infected with Echinococcus spp. worms. The worms were sent to the European Union Reference Laboratory (EURLP) for species/genotype identification. The sequences analyses of nad2 and nad5 genes allowed us to identify the tapeworms as Echinococcus canadensis, genotype G7b. In November 2023, a parasitic liver cyst was observed during the post-mortem examination of a mouflon from the same area of the dog's finding. The cyst sample was also referred to EURLP for identification and comparison with tapeworms previously collected from the dog. The sequences analysis of cox1 gene allowed to identify the cyst as E. granulosus sensu stricto, genotype G1. The finding of 2 different species of E. granulosus s.l. in a limited area raises epidemiological questions on the origin of the samples: whether distinct transmission cycles are present or a recent introduction event have occurred. From a public health perspective, it will be essential to conduct further molecular epidemiology studies to clarify the recent transmission dynamics of Echinococcus species in Cyprus.

Cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus sensu lato (s.l.), remains a significant zoonotic parasitic infection worldwide. This study provides a preliminary investigation of the nuclear Hsp90 gene diversity within the E. granulosus s.l. complex to evaluate its potential use for species identification. Forty-nine DNA samples of the G1 genotype from human and animal CE cysts, two G3 genotype samples, one of Echinococcus ortleppi , four of Echinococcus canadensis (G7), and four samples of other Taenia species ( Echinococcus multilocularis , Taenia hydatigena , Taenia pisiformis , and Taenia ovis ) were analyzed. Four primer pairs were designed to amplify the Hsp90 gene, followed by PCR amplification, DNA sequencing, and phylogenetic analysis. Successful amplification and sequencing of nearly the entire Hsp90 gene revealed a single nucleotide polymorphism (SNP) at position 222 conserved across all genotypes. Notably, significant genetic variations were observed between E. ortleppi (G5 genotype) and E. canadensis (G7 genotype) compared to E. granulosus sensu stricto (G1 and G3 genotypes). Phylogenetic analysis confirmed clustering consistent with established taxonomic relationships, with G1 and G3 forming a cluster, and G5 and G7 forming a distinct group. The findings suggest that the nuclear Hsp90 gene could be used as an additional marker for species-level differentiation within the E. granulosus s.l. complex.

Background Echinococcosis is a zoonotic parasitic disease causing serious health problems in both humans and animals in different endemic regions across the world. There are two different forms of human echinococcosis: Cystic Echinococcosis (CE) and Alveolar Echinococcosis (AE). CE is caused by the larval stage of Echinococcus granulosus sensu lato and AE by the larval stage of Echinococcus multilocularis . Geographically, CE is universally distributed, while AE is prevalent in the northern hemisphere. Although the disease is endemic in neighboring countries (China, Iran and India) of Pakistan, there are limited reports from that country. Besides, there are no comprehensive data on the genotyping of Echinococcus species in humans based on sequence analysis. This study aimed to detect the presence of human CE and to identify Echinococcus spp. in human isolates through genetic characterization of hydatid cysts in the Punjab Province of Pakistan. Methods Genetic analysis was performed on 38 human hydatid cyst samples collected from patients with echinococcosis using mitochondrial cytochrome c oxidase subunit 1 ( cox 1), cytochrome b ( cytb ) and NADH subunit 1 ( nad 1). Patient data including age, epidemiological history, sex, and location were obtained from hospital records. Results According to the sequence analysis we detected E. granulosus sensu stricto ( n  = 35), E. canadensis (G6/G7) ( n  = 2), and E. multilocularis ( n  = 1). Thus, the majority of the patients (92.1%, 35/38) were infected with E. granulosus s.s. This is the first molecular confirmation of E. canadensis (G6/G7) and E. multilocularis in human subjects from Pakistan. Conclusions These findings suggested that E. granulosus s.s. is the dominant species in humans in Pakistan. In addition, E. canadensis (G6/G7) and E. multilocularis are circulating in the country. Further studies are required to explore the genetic diversity in both humans and livestock.

Tapeworms of the species complex of Echinococcus granulosus sensu lato (s. l.) are the cause of a severe zoonotic disease – cystic echinococcosis, which is listed among the most severe parasitic diseases in humans and is prioritized by the World Health Organization. A stable taxonomy of E. granulosus s. l. is essential to the medical and veterinary communities for accurate and effective communication of the role of different species in this complex on human and animal health. E. granulosus s. l. displays high genetic diversity and has been divided into different species and genotypes. Despite several decades of research, the taxonomy of E. granulosus s. l. has remained controversial, especially the species status of genotypes G6–G10. Here the Bayesian phylogeny based on six nuclear loci (7387 bp in total) demonstrated, with very high support, the clustering of G6/G7 and G8/G10 into two separate clades. According to the evolutionary species concept, G6/G7 and G8/G10 can be regarded as two distinct species. Species differentiation can be attributed to the association with distinct host species, largely separate geographical distribution and low level of cross-fertilization. These factors have limited the gene flow between genotypic groups G6/G7 and G8/G10, resulting in the formation of distinct species. We discuss ecological and epidemiological differences that support the validity of these species.

European wolf populations have expanded their distribution ranges in recent decades and their spatial behaviour may contribute to the spread of Echinococcus spp. across Europe. In this study, the occurrence of Echinococcus granulosus sensu lato was monitored in the faecal samples from grey wolves from areas covering the substantial part of their range in Slovakia. Samples were collected from several geographical areas, including Poloniny National Park in the easternmost part of the country, as well as Muráň Plateau National Park, the Vepor Mountains and the Poľana Protected Landscape Area in central Slovakia. The overall occurrence of the parasite in 112 samples collected between 2014 and 2017 was 5.4%. Echinococcus canadensis (genotype G7), the main causative agent of human cystic echinococcosis in Central Europe, was confirmed in all six positive faeces through sequence analysis of the mitochondrial 12S rRNA gene. Geographically, five positive samples were derived from the Vepor Mountains and one from the Poľana Protected Landscape Area. The occurrence of E. canadensis G7, the primary cause of human cystic echinococcosis in Central Europe, in wolves in Slovakia indicates its presence in a sylvatic cycle, which may contribute to sustaining its circulation within the country.

Echinococcus granulosus sensu stricto ( s.s. ) is the primary cause of cystic echinococcosis (CE), a parasitic disease affecting humans and livestock with significant health and economic impacts. Previous studies on this parasite relied on mitochondrial DNA to classify its genotypes and understand its genetic diversity. However, these studies cannot capture the full complexity of its evolutionary dynamics and adaptation strategies. Our research employs comprehensive genome-wide sequencing, offering a more nuanced view of its genetic landscape. We discovered that cross-fertilization appears to be a prevalent reproductive strategy in the hermaphroditic E. granulosus , underpinning the observed deep mitochondrial divergence between genotypes G1 and G3, as well as gene flow among populations. The transmission history of E. granulosus s.s. in China and its widespread genetic mixing were likely facilitated by the migrations of nomadic peoples. Furthermore, we identified genes under balancing selection, including the gene involved in the uptake of host bile acids, which play a crucial role in the parasite’s survival and development, potentially offering new targets for intervention. Our research advances the understanding of the genetic diversity and evolutionary strategies of E. granulosus , laying the foundation for improved control measures of CE.

Background Cystic echinococcosis (CE) is a serious tapeworm infection caused by Echinococcus granulosus ( sensu lato ) which infects a wide range of animals and humans worldwide. Despite the millions of livestock heads reared in Pakistan, only a few reports on CE prevalence and even fewer on the genetic diversity are available for the country. Meanwhile, the available reports on the genetic diversity are predominantly based on short sequences of the cox 1 gene. Methods To close this knowledge gap, this study was designed to investigate the genetic diversity and population structure of Echinococcus spp. in Pakistan using the complete mitochondrial cytochrome c oxidase subunit 1 ( cox 1) and NADH dehydrogenase subunit 1 ( nad 1) genes. Results Based on BLAST searches of the generated cox 1 and nad 1 gene sequences from a total of 60 hydatid cysts collected from cattle ( n  = 40) and buffalo ( n  = 20), 52 isolates were identified as E . granulosus ( s.s. ) (G1, G3) and 8 as E . ortleppi (G5). The detection of the G5 genotype represents the first in Pakistan. The phylogeny inferred by the Bayesian method using nucleotide sequences of cox 1 -nad 1 further confirmed their identity. The diversity indices indicated a high haplotype diversity and a low nucleotide diversity. The negative values of Tajima’s D and Fu’s Fs test demonstrated deviation from neutrality suggesting a recent population expansion. Conclusions To the best of our knowledge, this report described the genetic variation of E. granulosus population for the first time in Pakistan using the complete cox 1 and nad 1 mitochondrial genes and confirms E. ortleppi as one of the causative agents of CE among livestock in Pakistan. While this report will contribute to baseline information for CE control, more studies considering species diversity and distribution in different hosts across unstudied regions of Pakistan are highly needed.