Cystic Echinococcosis (CE) is a zoonotic disease caused by Echinococcus granulosus sensu lato. Diagnosing CE primarily relies on imaging techniques, and there is a crucial need for an objective laboratory test to enhance the diagnostic process. Today, cell-free DNAs (cfDNAs) have gained importance regarding their biomarker potential. This study aims to investigate the diagnostic capabilities of different cfDNA targets (Echinococcus-specific repeat sequences (mgs-4 and mgs-12) and partial fragment of repetitive sequence (EG1 Hae III)) and evaluate their diagnostic effectiveness when compared to a frequently used commercial E.granulosus-specific IgG ELISA. Seventy-six confirmed hepatic CE patients and healthy controls were included in the study. The EG1 Hae III region was assessed using nested PCR, whereas real-time PCR was employed to investigate other cfDNA targets. Analysis of the cfDNA-targeted tests indicated that mgs-4 demonstrated the highest diagnostic efficacy in distinguishing CE patients from healthy controls, achieving a sensitivity of 60.5% (p = 0.002). Combining ELISA with the mgs-4 target led to an increased sensitivity of 72.4% for distinguishing between CE patients and the control group. The sensitivity rates for ELISA and the three cfDNA targets varied among the groups. Active CE patients showed sensitivity rates of 52.9%, 52.9%, 23.5%, and 52.9% for ELISA, mgs-4, mgs-12, and EG1 Hae III assays, respectively. In contrast, inactive cyst patients displayed sensitivity rates of 21.4%, 66.7%, 19%, and 42.9% for the corresponding assays. The mgs-4, either alone or in combination with ELISA, demonstrated notably higher sensitivity values for CE diagnosis in all group comparisons compared to serology.

Cestodiases, like echinococcoses and cysticercoses, represent a global health problem. Currently available anthelmintics, as benzimidazoles and praziquantel, have limited effectiveness against these cestodiases, creating a demand for the identification of new and more effective drugs. Here, the potential of statins (simvastatin and fluvastatin), for repositioning as novel anthelmintic is explored. Statins are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, a main enzyme in the mevalonate pathway, which is vital for the synthesis of non-steroidal isoprenoids and the maintenance of normal cell functioning. A survey for HMG-CoA reductase encoding genes showed that they are present in a single copy in the genomes of parasitic helminths and their mammal hosts. Sequence alignments and phylogenetic analyses showed 20–95% overall identities among ortholog HMG-CoA reductases, with special conservation of their catalytic domains. The HMG-CoA reductase 3D-structure was predicted for orthologs from 3 cestodes of medical importance (Echinococcus multilocularis, Echinococcus granulosus sensu lato, and Taenia solium), and from a model cestode species (Mesocestoides corti). Molecular docking between cestode HMG-CoA reductase orthologs with simvastatin demonstrated that the Arg, Ser, Lys, and Glu residues in conserved positions of the active site interact with this drug, similarly to the interactions predicted for the human reference ortholog enzyme. Furthermore, in vitro assays demonstrated that simvastatin produced a significant reduction of M. corti viability, being able to reduce 100% of parasite viability at 150 μm. Fluvastatin was also assessed showing a lower, although significant anthelmintic effect. The predicted overall structures and interactions together with in vitro assays suggest that cestodes HMG-CoA reductases are inhibited by simvastatin, being a potential therapeutic target for the repurposing of simvastatin as anthelmintic drug. Furthermore, these results pave the way for the in vivo evaluation of the potential effects of simvastatin on cestode larvae.

This study presents the first detection of Taenia omissa metacestodes in guanaco (Lama guanicoe) within the Chilean Patagonia, marking the southernmost record of natural infection in an intermediate host on the continent. Taenia omissa was found in the continental part of the Magallanes region where the top predators are pumas (Puma concolor). Conversely, all metacestodes found in guanacos collected from Tierra del Fuego Island, where no pumas exist, were identified solely as Echinococcus granulosus sensu stricto. Additionally, this research highlights a tissue preference of T. omissa for liver, contrasting with E. granulosus, which predominantly affects the lungs in guanacos. We also report the infection of T. pisiformis in 1 guanaco. Our findings emphasize the need for accurate identification of metacestodes during meat inspection in an area where T. omissa and E. granulosus overlap. This research also contributes to increase the knowledge of parasite–host dynamics in wildlife and underscores the importance of considering broader spectrum intermediate hosts in the epidemiology of parasitic infections.

Cystic and alveolar echinococcosis are considered the second and third most significant foodborne parasitic diseases worldwide. The microscopic eggs excreted in the feces of the definitive host are the only source of contamination for intermediate and dead-end hosts, including humans. However, estimating the respective contribution of the environment, fomites, animals or food in the transmission of Echinococcus eggs is still challenging. Echinococcus granulosus and E. multilocularis seem to have a similar survival capacity regarding temperature under laboratory conditions. In addition, field experiments have reported that the eggs can survive several weeks to years outdoors, with confirmation of the relative susceptibility of Echinococcus eggs to desiccation. Bad weather (such as rain and wind), invertebrates and birds help scatter Echinococcus eggs in the environment and may thus impact human exposure. Contamination of food and the environment by taeniid eggs has been the subject of renewed interest in the past decade. Various matrices from endemic regions have been found to be contaminated by Echinococcus eggs. These include water, soil, vegetables and berries, with heterogeneous rates highlighting the need to acquire more robust data so as to obtain an accurate assessment of the risk of human infection. In this context, it is essential to use efficient methods of detection and to develop methods for evaluating the viability of eggs in the environment and food.

Cystic echinococcosis (CE), caused by the larval stage of the cestode Echinococcus granulosus, is one of the most widespread zoonoses in Mediterranean countries. Baiting not-owned dogs with praziquantel (PZQ), due to their key role in the maintaining the transmission of CE, currently appears to be the most effective way to limit the transmission of CE, as well as an important aspect to introduce for the control of this parasitic disease. Therefore, this study aims to test 3 types of PZQ-based baits by evaluating different parameters (integrity over time, attractiveness and palatability for dogs, and mechanical resistance after release to different altitudes) and the bait acceptance in field by target animals, i.e. not-owned dogs, by using camera traps. The double PZQ-laced baits (with a double layer of highly palatable chews) showed the greatest resistance in the environment while also preserving the attractiveness and palatability up to 10 days, also withstood heights of 25 m, thus resulting as the most suitable also for drone delivery. The results on the field showed that most of the baits were consumed by not-owned dogs (82.2%), while the remaining were consumed by wild boars (8.9%), foxes (6.7%), badgers (1.1%) and hedgehogs (1.1%), confirming the specific and high attractiveness of the double PZQ-laced baits for the target population and highlights how an anthelmintic baiting programme may be a viable tool for the management of E. granulosus among free-ranging dog populations in endemic rural areas.

Most human cystic echinococcosis (CE) cases worldwide are attributed to Echinococcus granulosus sensu stricto (s.s), followed by the G6 and G7 genotypes. While E. granulosus s.s. has a cosmopolitan distribution, the G6 genotype is restricted to areas where camels and goats are present. Goats are the primary livestock in the Neuquén province in Argentina where the G6 genotype has been reported to be responsible for a significant percentage of CE human cysts genotyped. In the present study, we genotyped 124 Echinococcus cysts infecting 90 CE-confirmed patients. Echinococcus granulosus s.s. was identified in 51 patients (56.7%) with 81 cysts and the G6 genotype in 39 patients (43.3%) harbouring 43 cysts. Most CE cases ≤18 years were male suggesting pastoral work could be a risk factor for the infection. Echinococcus granulosus s.s. was significantly found more frequently in the liver (32/51 patients) and the G6 genotype in the lungs and extrahepatic localizations (27/39). The patients infected with E. granulosus s.s., presented up to 6 cysts while patients infected with G6 presented a maximum of 2. The diameter of lung cysts attributed to E. granulosus s.s. was significantly larger compared to lung cysts from G6. Following the WHO ultrasound classification of liver cysts, we observed inactive cysts in 55.6% of G6 cysts and only 15.3% of E. granulosus s.s cysts. In conclusion, we provide evidence of differences in clinical aspects of CE caused by E. granulosus s.s. and the G6 genotype of E. granulosus s.l. complex infecting humans.

The EG95 recombinant vaccine is protective against cystic echinococcus in animal intermediate hosts. Preparation of the existing, registered EG95 vaccines involves semi-purification of the vaccine protein, adding to the cost of production. Truncation of the EG95 cDNA, shortening both the amino and carboxy-termini of the protein, leads to high levels of recombinant protein expression. The recombinant EG95 protein was prepared as a bacterin from clarified, whole bacterial lysate, and used in a vaccine trial in sheep against an experimental challenge infection with Echinococcus granulosus eggs. The EG95 bacterin was found to induce 98% protection. Use of this in a new generation EG95 vaccine would simplify production, facilitate new sources of the vaccine and potentially enhance uptake of vaccination in control of E. granulosus transmission.

Cystic echinococcosis (CE) occurs in the intermediate host's liver, assuming a bladder-like structure surrounded by the host-derived collagen capsule mainly derived from activated hepatic stellate cells (HSCs). However, the effect of CE on liver natural killer (NK) cells and the potential of transforming growth factor-β (TGF-β) signalling inhibition on alleviating CE-related liver damage remain to be explored. Here, by using the CE-mouse model, we revealed that the inhibitory receptors on the surface of liver NK cells were up-regulated, whereas the activating receptors were down-regulated over time. TGF-β1 secretion was elevated in liver tissues and mainly derived from macrophages. A combination of TGF-β signalling inhibitors SB525334 and pirfenidone could reduce the expression of TGF-β1 signalling pathway-related proteins and collagen production. Based on the secretion of TGF-β1, only the pirfenidone group showed a depressing effect. Also, the combination of SB525334 and pirfenidone exhibited a higher potential in effectively alleviating the senescence of the hepatocytes and restoring liver function. Together, TGF-β1 may be a potential target for the treatment of CE-associated liver fibrosis.

Cystic echinococcosis (CE) is a neglected tropical disease, caused by metacestode (larval) form of the Echinococcus granulosus sensu lato (sl) in humans. MicroRNAs (miRNAs) are small, stable, tissue-specific RNA molecules encoded by the genome that are not translated into proteins. Circulating miRNA expression profiles vary in health and disease. The aim of this study is to determine the altered cellular pathways in CE by comparing the miRNA profiles of controls and CE patients with active or inactive cysts. Following abdominal ultrasonography (US) examination, 20 patients diagnosed with active CE (CE1, CE2, CE3a and CE3b) or inactive CE (CE4 and CE5) and three healthy controls were included in the study. The expression profiles of 372 biologically relevant human miRNAs were investigated in serum samples from CE patients and healthy controls with miScript miRNA HC PCR Array. Compared with the control group, expression of 6 miRNAs (hsa-miR-4659a-5p, hsa-miR-4518, hsa-miR-3977, hsa-miR-4692, hsa-miR-181b-3p, hsa-miR-4491) and one miRNA (hsa-miR-4687-5p) were found to be downregulated in CE patients with active and inactive cysts, respectively (p < 0.05). For downregulated miRNAs in this study, predicted targets were found to be associated mainly with cell proliferation, apoptosis, cell-cell interactions and cell cycle regulation. Further studies in this direction may elucidate the pathogenesis of human CE and the relationship between CE and other pathologies.

Hydatidosis is a potential zoonotic helminthic disease affecting a broad spectrum of mammals, including humans, worldwide. The current review was conducted to investigate the genotypic status and prevalence of hydatid disease in camels across the world. For the purpose of the study, the articles addressing the worldwide prevalence of hydatidosis in camels were searched in several English language databases. The search process resulted in the inclusion of 122 papers. Based on the data presented in the reviewed articles, the pooled prevalence of hydatid disease in camels across the world was measured at 23.75% (95% CI 20.15–27.55). Moreover, the subgroup analysis demonstrated significant differences in the overall prevalence of hydatidosis among camels based on year, geographic area, climate parameters, camel population, gender, infected organ, fertility rate of the cyst and laboratory diagnostic technique. Furthermore, the Echinococcus granulosus genotypes identified in camels with hydatidosis included G1, G2, G3, G1–G3, G5, G6, G7, G6–G7 and G6–G10, with G6 being the most common genotype throughout the world. The data obtained from the current study are central to the better conceptualization of the biological and epidemiological characteristics of E. granulosus s.l. genotypes around the world, which can be helpful in the planning and adoption of more comprehensive control strategies.

In this study, we evaluated the efficacy, expressed as a mean weight decrease of the whole echinococcal cyst mass, of novel benzimidazole salt formulations in a murine Echinococcus granulosus infection model. BALB/c mice were intraperitoneally infected with protoscoleces of E. granulosus (genotype G1). At 9 months post-infection, treatment with albendazole (ABZ), ricobendazole (RBZ) salt formulations, and RBZ enantiomer salts (R)-(+)-RBZ-Na and (S)-(−)-RBZ-Na formulations were initiated. Drugs were orally applied by gavage at 10 mg kg−1 body weight per day during 30 days. Experimental treatments with benzimidazole sodium salts resulted in a significant reduction of the weight of cysts compared to conventional ABZ treatment, except for the (S)-(−)-RBZ-Na enantiomer formulation. Scanning electron microscopy and histological inspection revealed that treatments impacted not only the structural integrity of the parasite tissue in the germinal layer, but also induced alterations in the laminated layer. Overall, these results demonstrate the improved efficacy of benzimidazole salt formulations compared to conventional ABZ treatment in experimental murine cystic echinococcosis.

Echinococcus granulosus is the causative agent of cystic echinococcosis, which has serious impacts on human and/or animal health, resulting in significant economic losses. Echinococcus granulosus comprises a number of intra-specific variants or strains at the genetic level. In Saudi Arabia, few studies were performed on genetic variations in Echinococcus species. Therefore, the present study aimed to investigate the phenotypic and genetic characterization of hydatid cysts harboured by sheep and camels in Al-Madinah Al-Munawarah. Samples of hydatid cysts were collected from local sheep (n = 25) and camels (n = 8). The morphological criteria of protoscoleces were investigated. To investigate the molecular characterization, random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR), single-stranded conformation polymorphism (SSCP) were carried out. DNA was extracted from individual fertile cysts and subjected to RAPD-PCR analysis (using five arbitrary primers) and PCR amplification of cytochrome c oxidase I (cox1) and 12S ribosomal ribonucleic acid (12S rRNA) genes. The PCR products were subjected to SSCP analysis for genetic discrimination in E. granulosus isolates. In addition, partially sequencing of the mitochondrial DNA cox1 genes was achieved for assessing the phylogenetic positions of collected isolates using some global published sequence data of cox1 genes. The rostellar hooks of camel and local sheep isolates show remarkable variability in their dimensions. Five distinct SSCP patterns were identified in the 12S rRNA gene, showing intraspecific variations in E. granulosus of camels and local sheep. Sequencing of (cox1) genes of both local sheep and camels exhibit high similarity with those of the same gene (E. granulosus sensu stricto) published in NCBI BLAST.

Larval stage of genus Echinococcus is the causing agent for the zoonotic infection which is life threatening known as Echinococcosis. The purpose of this study was the identification, molecular analysis and characterization of Echinococcus spp. in sheep and cattle. The sampling was done from slaughterhouse of Elazig, Turkey. A total of 85 isolates (sheep, n = 19 and cattle, n = 66) have been collected after slaughtering. Following the gDNA isolation and PCR products of mt-CO1 gene (446 bp) of all the samples were sequenced. Out of 85 isolates, 84 were recognized as Echinococcus granulosus sensu stricto and one sheep isolate was found as Echinococcus canadensis (G6/G7 ) which is identified for the first time in Turkey. However, single nucleotide polymorphism has been observed not only in samples of different animals but also in samples collected from the same cattle. Six liver and three lung hydatid cysts have been detected in cattle. Although no nucleotide differences have been observed in the liver samples, there was single nucleotide polymorphism (C→T) in 40th nucleotide of two lung cysts. As a result of haplotype analysis, 16 haplotypes of E. granulosus s.s. were detected in 66 cattle isolates whereas 7 haplotypes of E. granulosus s.s. were identified in 19 sheep samples.

Cystic echinococcosis (CE) is a zoonotic disease caused by a complex of species known as Echinococcus granulosus sensu lato. CE is endemic in Argentina, Chile, Peru, Uruguay and the South part of Brazil. In contrast, little is known regarding the presence of CE in Bolivia. In this study, 35 cysts isolated from livestock (mostly from the Department of La Paz) and 3 from humans (La Paz, Oruro and Potosi) were genetically characterized analysing the sequence of the cox1 gene (1609 bp). In total, 30 cysts (from La Paz, Cochabamba and Beni) were characterized as E. granulosus sensu stricto (3 fertile and 4 non-fertile cysts from sheep, 8 fertile and 12 non-fertile cysts from cattle and 3 fertile cysts from humans). A detailed analysis of the cox1 haplotypes of E. granulosus s.s. is included. Echinococcus ortleppi (G5) was found in 5 fertile cysts from cattle (from La Paz and Cochabamba). Echinococcus intermedius (G7) was identified in 3 fertile cysts from pigs (from Santa Cruz). Additionally, E. granulosus s.s. was detected in 4 dog faecal samples, while E. ortleppi was present in other two dog faecal samples. The implications of these preliminary results in the future implementation of control measures are discussed.

Echinococcus granulosus sensu stricto is regarded to have the highest zoonotic potential of all Echinococcus taxa. Globally, human infection due to this species constitutes over 88.44% of the total cystic echinococcosis (CE) burden. Here, we report a CE infection in a Nigerian camel caused by E. granulosus G1 genotype. To the best of our knowledge, this report is the first encounter of the G1 genotype in the West Africa sub-region where the G6 genotype is reportedly prevalent, suggesting that the epidemiology of this highly zoonotic group could have a wider host range and distribution in the sub-region, and emphasizes the need for further investigation into the genetic diversity of Echinococcus spp. in Nigeria and across the sub-region.

Despite several studies conducted to determine the genotypes of cystic echinococcosis (CE) agents in humans and other intermediate hosts, the relationship between parasite genotype and clinical presentation of hydatidosis is yet to be well defined. The aim of this study was to compare the genotypes/haplotypes of CE agents of human extra-hepatopulmonary hydatid cysts and common hydatid cysts of the liver. A comparative analysis was carried out between partial cox1 sequences of ten extra-hepatopulmonary hydatid cysts, two liver cysts and reliable sequences from the GenBank database. All the studied hydatid cysts had the Echinococcus granulosus sensu stricto (G1-G2-G3) genotypes. The liver CE cysts were caused by common G1 genotype, while six of the extra-hepatopulmonary cysts had genotypes different from common G1 cysts. The sequences of these six isolates were identical to the G2 and G3 genotypes of E. granulosus sensu stricto; the kidney and peritoneum cysts and most of the brain cysts were identified as G2 genotype, while G3 genotype was only reported in a cyst belonging to the pelvic region. Given the observed differences between the sequences of hydatid cysts, it seems that the replacement of hydatid cysts in organs other than the liver and lungs can be related to their genotypes and probably intra-genotypic characteristics. It was hypothesized that in each geographic area, less frequent genotypes were likely to be more consistent with placement in the host's unusual organs.

Cystic echinococcosis caused by Echinococcus granulosus is a major zoonosis of public health significance in the Patagonian region of Argentina. This investigation sought to test the hypothesis that the persistence and dispersion of the parasite eggs can be explained by physical and meteorological parameters along with final host infection and behaviour. This observational study was carried out over a five-year period within an enclosure where two dogs harbouring a worm burden ranging from 100 to 1000 mature adult E. granulosus, as well as two uninfected dogs, had previously been kept for six months. Environmental canine faeces, topsoil, pond water, and sediment samples were examined to control for the presence of eggs and coproantigens of the parasite using microscope-based techniques and copro-ELISA plus copro-Western Blot tests. The parasite eggs were detected up to 41 months later in faeces from infected dogs, soil and sediment, and coproantigen tests remained positive for up to 70 months in faeces. Overall, parasite eggs were found within a maximum distance of 115 m from the contaminated dog faeces deposition site. Our findings indicate that under Patagonian environmental conditions, egg persistence and dispersion seem to be related to the worm burden and habits of the infected dog, to prevailing wind direction and to the existence of low bushes as well as natural bodies of water. The present study is the first to provide direct evidence of interaction between bioclimatic conditions and E. granulosus egg dispersion under Patagonian field conditions.

Diagnosis of cystic echinococcosis (CE) is at present mainly based on imaging techniques. Serology has a complementary role, partly due to the small number of standardized and commercially available assays. Therefore we examined the clinical performance of the SERION ELISA classic Echinococcus IgG test. Using 10 U/ml as a cut-off point, and serum samples from 50 CE patients and 105 healthy controls, the sensitivity and specificity were 98.0% and 96.2%, respectively. If patients with other infectious diseases were used as negative controls, the specificity decreased to 76.9%, which causes poor positive predictive values. However, if results between 10 and 15 U/ml are classified as indecisive, the specificity of positive results (≥15 U/ml) increased to 92.5% without greatly affecting the sensitivity (92.0%). Using this approach in combination with imaging studies, the SERION ELISA classic Echinococcosis IgG test can be a useful aid in the diagnosis of CE.

Cystic echinococcosis (CE) is a severe parasitic disease caused by the species complex Echinococcus granulosus sensu lato. Human infections are most commonly associated with E. granulosus sensu stricto (s.s.), comprising genotypes G1 and G3. The objective of the current study was to provide first insight into the genetic diversity and phylogeography of genotype G3. Despite the epidemiological importance of the genotype, it has remained poorly explored due to the ambiguity in the definition of the genotype. However, it was recently demonstrated that long sequences of mitochondrial DNA (mtDNA) provide a reliable method to discriminate G1 and G3 from each other. Therefore, we sequenced near-complete mtDNA of 39 G3 samples, covering most of the known distribution range and host spectra of the genotype. The phylogenetic network revealed high genetic variation within E. granulosus s.s. G3 and while G3 is significantly less prevalent worldwide than G1, the genetic diversity of both of the genotypes is equally high. We also present the results of the Bayesian phylogeographic analysis, which yielded several well-supported diffusion routes of genotype G3 originating from Turkey and Iran, suggesting the Middle East as the origin of the genotype.

Echinococcus granulosus is an important zoonotic parasite that is distributed worldwide. The EG95 vaccine was developed to assist with control of E. granulosus transmission through the parasite's livestock intermediate hosts. The vaccine is based on a recombinant antigen encoded by a gene which is a member of a multi-gene family. With the recent availability of two E. granulosus draft genomes, we sought to map the eg95 gene family to the genomes. We were unable to map unequivocally any of the eg95 gene family members which had previously been characterized by cloning and sequencing both strands of genomic DNA fragments. Our inability to map EG95-related genes to the genomes has revealed limitations in the assembled sequence data when utilized for gene family analyses. This study contrasts with the expectations expressed in often high-profile publications describing draft genomes of parasitic organisms, highlighting deficiencies in currently available genomic resources for E. granulosus and provides a cautionary note for research which seeks to utilize these genome datasets.