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is causative agent of cystic echinococcosis (CE), which has a cosmopolitan distribution. The current methods for the treatment of human CE include surgery. Therefore, the development of new scolicidal agents with low side effects and more efficacies is an urgent need. The present study aimed to compare the scolicidal efficacies of silver, iron, copper, silica and zinc oxide nanoparticles (NPs) against hydatid cyst protoscolices in vitro. Hydatid cysts of sheep liver and lung were collected. The cyst fluid containing protoscolices was aspirated aseptically. The scolicidal activities of the silver, iron, copper, silica and zinc nanoparticles (Ag-NP, Fe-NP, Cu-NP, Si-NP and Zn-NP) were tested at different concentrations of 0.25, 0.5 and 1 mg/mL following 10, 30 and 60 min of incubation in triplicate. Viability of protoscolices was confirmed by 0.1% eosin staining. Results showed that Ag-NPs at all concentrations tested had the highest scolicidal effect. Ag-NPs at 1 mg/mL concentration after 60 min of exposure time showed 80% mortality rate. Si-NPs had the high scolicidal activity at 1 mg/mL concentration (52.33%), Cu-NPs at 0.5 mg/mL concentration (41%), Fe-NPs at 1mg/mL concentration (28%) and Zn-NPs at concentration of 1mg/mL after 60 mins (15.67%). The findings of the present study showed that Ag-NPs, Fe-NPs, Cu-NPs, Si-NPs and Zn-NPs had potent scolicidal effects and that Ag-NPs are recommended as effective scolicidal agents. However, further in vivo studies are required to evaluate the efficacy of these nanoparticles.

Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans can become accidental but dead-end hosts. The aim of this study is to use EmsB typing on E. multilocularis isolates from human AE cases and local animals such as foxes and rodents. In this study, retrospective EmsB typing was performed on 39 samples, including 11 tissue samples from 10 patients, 18 fecal swabs from foxes, and 10 tissue samples from rodents. A dendrogram was created to determine the EmsB profiles present. The results showed that all the rodent samples were associated with the EmsB P1 profile (10/10), while the human and fox samples shared the EmsB profile P1 (5/11 humans and 8/18 foxes), a profile near P4 (2/11 humans and 3 foxes), and a profile near P8 (1/11 humans and 1/18 foxes). The study demonstrates that the same EmsB profiles circulate among humans and animals, confirming that wildlife reservoirs play a key role in transmission.

Cystic Echinococcosis (CE) is a zoonotic disease caused by Echinococcus granulosus sensu lato , forming cysts in ruminants and humans with major health and economic impacts. The immune response to CE cysts is complex, with fertility linked to the host’s inflammatory reaction. This study examines γδ T cell distribution and role within the adventitial layer of non-fertile CE cysts in cattle, including cases co-infected with the trematode Fasciola hepatica (FH), a known immune response modulator. Using immunohistochemistry and double immunofluorescence, we observed γδ T cells dispersed in the adventitial layer, enriched in inflammatory zones. Co-infected cases (CE + FH+) showed a reduced γδ T cell proportion among CD3+ T cells compared to non-coinfected cases, suggesting an immunoregulatory effect of FH. Our findings align with prior studies showing γδ T cell recruitment in granulomatous diseases in ruminants but reveal that co-infection alters this response. This study provides the first detailed characterization of γδ T cells in cattle CE cysts, emphasizing their potential role in granulomatous immune responses. It highlights the need for further research into mechanisms influencing CE cyst fertility and immune modulation in helminth co-infections, advancing our understanding of host-pathogen interactions and informing disease management strategies.

Accumulating evidence has shown that adipose-derived stem cells (ADSCs) have the potential to differentiate into hepatic lineages, which are ideal engraftments for tissue-engineered repair. In this study, we investigated the potential of transplanted induced hepatocyte-like cells (iHEPs) in treating hepatic alveolar echinococcosis and describe an efficient three-step protocol for the generation of iHEPs in vitro from ADSCs. The expression of hepatocyte lineage markers was assessed and iHEPs function was evaluated by Periodic acid-Schiff staining. iHEPs were intravenously transplanted into mice infected with Echinococcus multilocularis . Histopathological analysis and liver function tests were used to assess therapeutic effects. The iHEPs exhibit morphological features and a glycogen storage function similar to those of mature hepatocytes and demonstrate an upregulation in hepatic gene programs with increasing induction time. Following transplantation, iHEPs were observed surrounding the metacestode lesions in the liver parenchyma of E. multilocularis- infected mice. iHEPs transplantation effectively restored liver function and improved liver injury in the infected mice. Additionally, we observed significant activation of the Wnt/β-catenin signaling pathway in the livers of infected mice transplanted with iHEPs. Our results provide evidence that iHEPs transplantation can alleviate E. multilocularis -induced liver injury, potentially creating new avenues for treating liver injury in end-stage hepatic alveolar echinococcosis.

Alveolar (AE) and cystic echinococcosis (CE) in humans are caused by the metacestode of the tapeworms Echinococcus multilocularis and Echinococcus granulosus sensu lato (s.l.). Immunohistochemistry with the monoclonal antibodies (mAb) Em2G11, specific for AE, and the mAb EmG3, specific for AE and CE, is an important pillar of the histological diagnosis of these two infections. Our aim was to further evaluate mAb EmG3 in a diagnostic setting and to analyze in detail the localization, distribution, and impact of small particles of Echinococcus multilocularis (spems) and small particles of Echinococcus granulosus s.l. (spegs) on lymph nodes. We evaluated the mAb EmG3 in a cohort of formalin-fixed, paraffin embedded (FFPE) specimens of AE (n = 360) and CE (n = 178). These samples originated from 156 AE-patients and 77 CE-patients. mAb EmG3 showed a specific staining of the metacestode stadium of E. multilocularis and E. granulosus s.l. and had a higher sensitivity for spems than mAb Em2G11. Furthermore, we detected spegs in the surrounding host tissue and in almost all tested lymph nodes (39/41) of infected patients. 38/47 lymph nodes of AE showed a positive reaction for spems with mAb EmG3, whereas 29/47 tested positive when stained with mAb Em2G11. Spegs were detected in the germinal centers, co-located with CD23-positive follicular dendritic cells, and were present in the sinuses. Likewise, lymph nodes with spems and spegs in AE and CE were significantly enlarged in size in comparison to the control group. mAb EmG3 is specific for AE and CE and is a valuable tool in the histological diagnosis of echinococcosis. Based on the observed staining patterns, we hypothesize that the interaction between parasite and host is not restricted to the main lesion since spegs are detected in lymph nodes. Moreover, in AE the number of spems-affected lymph nodes is higher than previously assumed. The enlargement of lymph nodes with spems and spegs points to an immunological interaction with the small immunogenic particles (spems and spegs) of Echinococcus spp.

Introduction Alveolar echinococcosis (AE), caused by the larval forms of Echinococcus multilocularis , is a zoonotic disease affecting the liver, lungs, lymph nodes, kidneys, brain, bones, thyroid, and other organs. Diagnosing AE in a non-endemic area is usually challenging. With the rapid development and increasing application of sequencing techniques in recent years, metagenomic next-generation sequencing (mNGS) has become a powerful tool for diagnosing rare infectious diseases. Case Presentation A 45-year-old woman was admitted to the hospital for the presence of pulmonary shadows for more than 3 months. The lung computed tomography (CT) at a local hospital revealed scattered solid and quasi-circular nodules in the left upper lobe, left lower lobe, right middle lobe, and right lower lobe. The largest nodule was located in the dorsal part of the right lung, measuring 2.0 × 1.7 × 1.5 cm. Moreover, abdominal CT revealed one space-occupying lesion each in the left and right lobes. The pathological analysis of the lung biopsy specimen revealed infiltration of lymphocytes, plasma cells, and eosinophils in the alveolar wall and interstitial area. No pathogenic bacteria were observed in the sputum smear and culture tests. There were no parasite eggs in the stool. The mNGS of the lung puncture tissue revealed 6156 sequence reads matching E. multilocularis ; thus, the condition was diagnosed as AE. Albendazole 400 mg was administered twice daily, and the patient was stable during follow-up. Conclusion This case emphasizes the role of mNGS in diagnosing AE. As a novel, sensitive, and accurate diagnostic method, mNGS could be an attractive approach for facilitating early diagnosis and prompt treatment of infectious diseases, especially when the infection was caused by rare pathogens.

The tapeworms Echinococcus granulosus and Echinococcus multilocularis cause two different clinical manifestations in humans: cystic echinococcosis (CE) and alveolar echinococcosis (AE), respectively. Both forms of echinococcosis manifest primarily in the liver, while other organs or tissues are less frequently affected. Simultaneous occurrence of CE and AE is extremely rare, and all previously reported patients exhibited affected livers, while simultaneous infection without liver involvement has not yet been described. Herein, we present an exclusively extrahepatic E. granulosus and E. multilocularis coinfection localized between the calf muscles of a patient. Due to progressive painful local swelling, an abscess was suspected, but there was no improvement after the administration of multiple courses of antibiotics. When imaging diagnostics suggested a parasitic origin of the two identified cystic lesions, positive serology for both species indicated a dual infection. Albendazole therapy was started, and extensive surgical excision was performed. Both species were confirmed using PCR and sequencing from intraoperative samples. The current case shows that coinfection without liver involvement can occur, even in patients from low-incidence regions, which should be considered in the differential diagnosis of patients with unusual clinical presentation.

Background Alveolar echinococcosis (AE), a fatal disease caused by Echinococcus multilocularis , often affects the liver, with tumor-like growth. However, the mechanism by which E. multilocularis evades host immune surveillance remains unclear. Methods We collected liver specimens from hepatic alveolar echinococcosis (HAE) patients and established a mouse model of E. multilocularis infection. Immunofluorescence staining and flow cytometry were performed to analyze programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte associated antigen 4 (CTLA-4) expression in human samples, while flow cytometry and quantitative real-time polymerase chain reaction (PCR) were performed for similar analyses in mouse samples. Cell proliferation and protoscolex (PSC) killing assays were designed to explore how E. multilocularis induces host immunosuppression. Results An inflammatory reaction band with high PD-1 and CTLA-4 expression was found in close liver tissue (CLT). The ratio of regulatory T cells (Tregs) was higher in CLT than in distant liver tissue (DLT), and Tregs in CLT tended to express higher levels of PD-1 and CTLA-4 than those in DLT from HAE patients. Echinococcus multilocularis -infected mice showed significantly elevated expression of PD-1 and CTLA-4 on splenocytes and peritoneal cells. PD-1/PD-L1 or CTLA-4 pathway blockade could relieve the immunosuppressive effects of Tregs from infected mice and enhance PSC killing by mouse splenocytes. Conclusions E. multilocularis regulated the function of T cells via the PD-1/PD-L1- and CTLA-4-dependent pathways and subsequently evaded host immune attacks. These findings provide insights for investigating the pathogenic mechanism of AE. Graphical Abstract

Background Cystic echinococcosis (CE) and alveolar echinococcosis (AE) are zoonotic diseases caused by the larval stages of Echinococcus granulosus and E. multilocularis . Prior studies have identified small particles from the outer layer of parasitic lesions termed spegs (from E. granulosus ) and spems (from E. multilocularis ), which appear to interact with the human immune system, leading to lymph node enlargement. Methods We analyzed lymph nodes from CE ( n  = 3) and AE ( n  = 3) patients by immunohistochemical staining for spegs/spems and CD57. Automated image analysis revealed a significantly increased number of CD57-positive cells in Echinococcus particle-positive germinal centers compared with speg-/spem-negative germinal centers. Double immunofluorescence staining was used to characterize CD57-positive cells with markers for CD2, CD3, CD4, CD8, PD-1, Granzyme B, Perforin, and T-cell intracellular antigen 1 (TIA1). Additionally, IgG4 and IgE immunostaining was performed on liver lesions and lymph nodes containing spegs and spems and compared with unaffected controls. Results Spegs and spems were found to accumulate in the germinal centers of the lymph nodes. Affected germinal centers showed a higher number of CD57-positive cells, which coexpressed CD2, CD3, CD4, and PD-1 but lacked cytotoxic markers (CD8, Granzyme B, Perforin, TIA1), identifying them as follicular helper T cells (Tfh). Furthermore, the speg- and spem-affected liver and lymph node sections exhibited significantly increased numbers of IgG4-, IgE-, and CD57-positive cells. Conclusions Spegs and spems are associated with changes in the host immune response, characterized by an increased accumulation of Tfh in germinal centers. This finding is accompanied by the presence of IgE and IgG4 plasma cells in the lymph node and the perilesional area, suggesting parasite-associated immunomodulatory processes in echinococcosis. Graphical Abstract

Alveolar echinococcosis caused by Echinococcus multilocularis is common parasitic disease among humans and animals in the northern hemisphere. Dogs, foxes, and other wild canids are definitive hosts, whereas small rodents play the role of intermediate hosts. In rare cases, after incidental ingestion of tapeworm eggs, dogs can become an intermediate host. The study describes briefly two cases of alveolar echinococcosis in dogs in Poland, including clinical management, diagnostic, treatment and molecular confirmation. Diagnostic procedures included laparotomy, cytology, histopathology and molecular analysis. Obtained sequences data were 100% homologous to E. multilocularis dehydrogenase subunit 1 gene sequences in GenBank®. To the authors' knowledge, alveolar echinococcosis has not been reported previously in a dog in Poland.