Cardiac echinococcosis, a multidisciplinary approach in the diagnosis and treatment of this rare entity: two case reports and literature review.

We present two case reports of cardiac echinococcosis. Case 1 was a 33-year-old woman with hepatic and cardiac echinococcosis. The parasitic cyst was located intramyocardially in the free wall of the left ventricle leading to cranial dislocation of the left circumflex coronary artery (LCx). The patient was successfully operated. Case 2 was a 28-year-old woman with hepatic and cardiac echinococcosis. The parasitic cyst was located in the left ventricular myocardium in the area of the apex and manifested clinically as paroxysms of ventricular tachycardia. The ultrasound study showed a 3.2×2.8 cm cyst dislocating the papillary muscles and causing moderate mitral regurgitation.Bulgaria ranks first in the European Union in terms of the number of echinococcosis patients. Although cardiac involvement is uncommon, occurring in only 0.5%-2% of cases, it can cause a wide range of clinical symptoms. Multimodal imaging is a key step in the management of patients with cardiac involvement.

Genetic characterization of human echinococcosis in Southern Punjab, Pakistan.

Introduction: Echinococcosis is a neglected tropical zoonotic infection that affects both the human and livestock populations. In Pakistan, the infection is long-standing, but data on its molecular epidemiology and genotypic characterization in the southern Punjab region are limited. The aim of the current study was the molecular characterization of human echinococcosis in southern Punjab, Pakistan. Methods: Echinococcal cysts were obtained from a total of 28 surgically treated patients. Patients' demographic characteristics were also recorded. The cyst samples were subjected to further processing to isolate DNA in order to probe the Nad1 and Cyt-b genes, followed by DNA sequencing and phylogenetic analysis for genotypic identification. Results: The majority of the echinococcal cysts were from male patients (60.7%). The liver was the most commonly infected organ (60.71%), followed by the lungs (25%), spleen (7.14%), and the mesentery (7.14%). Molecular and genotypic identification through sequencing and phylogenetic tree analysis showed that most of the cysts (24/28, 85.7%) were caused by the species Echinococcus granulosus sensu stricto (E. granulosus s.s.) (G1 and G3), followed by Echinococcus multilocularis (E. multilocularis) and Echinococcus canadensis (E. canadensis) (G6/G7) (3/28, 10.8%, and 1/28, 3.5%, respectively). Conclusion: The current study concluded that the majority of human infections were caused by E. granulosus s.s., followed by the E. multilocularis and E. canadensis species (G6/G7). Genotypic characterization among both human and livestock populations is needed to explore the genetic diversity of echinococcosis.

Diagnosis and surgical treatment of cysto-gastric fistula out of an hepatic hydatid cyst. Unusual case.

BACKGROUND: Hydatid cyst is an endemic zoonotic infection that annual incidence ranges from <1 to 200 per 100,000 individuals. The most common complication of hepatic hydatid cyst reported is rupture of the cysts, most commonly intrabiliary rupture. Direct rupture to hollow visceral organs is rarely seen. We describe here an unusual cystogastric fistula in a patient with liver hydatid cyst. CASE PRESENTATION: The 55-year-old male patient presented with right upper quadrant abdominal pain. After radiological imaging studies, the diagnose was of hydatid cyst involving the left lateral segment of the liver ruptured into the gastric lumen and resulted in a cystogastric fistula. Gastroscopy revealed that the cyst and its contents protruding from anterior wall to the gastric lumen. Partial pericystectomy and omentopexy were performed and the gastric wall was primarily repaired. There were no complications in the postoperative period and 3-month follow up. CONCLUSION: This case, to our knowledge, is the first reported case of cystogastric fistula surgically treated in a patient with liver hydatid cyst in the literature. Our clinical experience shows that, although it is a benign disease, complicated hydatid cysts should be evaluated in detail preoperatively, and after the detailed diagnostic work-up, surgical therapy might be planned individually for each case. KEY WORDS: Cysto-gastric fistula, Hydatid Cyst, Liver hydatidosis.

Mixed epithelial and stromal tumour of the kidney masquerading as hydatid cyst: a rare case report from the developing world.

Mixed epithelial and stromal tumour (MEST) is a rare entity with an incidence of 0.2% among renal cancers. It has strong predilection towards females with a 1:6 male to female ratio, the tumour is cystic with some solid component with biphasic proliferation of stromal and epithelial cells. The case of a 37-years-old female is presented with right lumbar pain since 3 months. The family history was unremarkable. The routine workup revealed mild neutrophilia and borderline Echinococcus antibody titres. Ultrasound revealed a complex cystic lesion with a solid component in the right kidney. CT scan with contrast confirmed a multiloculated mixed density lesion with daughter cysts arising from the middle lobe of the right kidney. Initial diagnosis of renal hydatid cyst was established and she underwent partial nephrectomy with excision of the cystic mass. Surprisingly the histopathology revealed mixed epithelial and stromal tumour.

Attitude and awareness of Algerian university students about cystic echinococcosis: A cross-sectional study.

Cystic echinococcosis is a neglected tropical parasitic disease affecting both humans and animals with considerable socioeconomic impact on affected pastoral and poor communities. CE is endemic in the Mediterranean basin, including Algeria posing a real threat for either animal and public health. This study was conducted to evaluate the level of awareness and knowledge of Algerian university students about this disease and to describe their attitude toward some risky practices. Results showed that despite that a high number of students (76.1%) have heard about CE, their level of knowledge is however medium (63.3%) especially among students of non medical and life sciences specialties. The gaps of knowledge concerns especially the life cycle of the parasite even they are aware about the association of dogs (43.7%) and sheep (59.4%) and considered that dogs are the main source of contamination of humans (58.1%) through contaminated foods (45%) or direct contact with their faeces (44.5%). Regarding their practice, a good level was shown since washing vegetables (99.2%) and washing hands after touching a dog (97.9%) and dogs deworming (82%) were common. These results highlight the need to improve knowledge about the transmission cycle of the parasite through awareness campaign targeting students which could be helpful in CE eradication.

Association between radical versus conservative surgery and short-term outcomes of hepatic cystic echinococcosis in Nyingchi, China: a retrospective cohort study.

BACKGROUND: Radical or conservative surgical treatment for hepatic Cystic Echinococcosis (hepatic CE) is controversial. We aimed to measure the association between radical surgery (RS) versus conservative surgery (CS) and short-term outcomes in our cohort. METHODS: Medical records of hepatic CE patients' demographic, clinical, radiological, operative and postoperative details who underwent surgical treatment between January 3, 2017 and January 3, 2018 at the Department of General Surgery, Nyingchi People's Hospital, Nyingchi, China, were retrieved and analyzed. The primary outcome was overall morbidity. The secondary outcomes included: (i) bile leakage; (ii) complications of lung, pleura, heart, liver, pancreas and biliary tract; (iii) incision infection and residual cavity abscess formation; (iv) anaphylactic reaction and shock; (v) tear of surrounding tissues; (vi) hospital and post-operative length of stay (LOS); (vii) length of surgery; (viii) blood loss during surgery. Multivariable logistic/linear regression models with various adjustment strategies for confounders were performed to evaluate the association. RESULTS: A total of 128 hepatic CE patients were included with 82 (64.1%) and 46 (35.9%) receiving CS and RS, respectively. After fully adjusted, RS was associated with 60% lower risk of overall complication (aOR 0.4; 95%CI, 0.2-0.9) and 0.6-h shorter surgical time (aß 0.4; 95%CI,-0.0-0.8) comparing to CS. However, RS was associated with more blood loss during surgery (aß 179.3; 95%CI, 54.2-304.5). CONCLUSION: To conclude, RS was associated with a 60% reduction in developing overall complication in the short term, but may result in more blood loss during surgery than CS.

Precordial pain induced by the isolated cardiac hydatid cyst in interventricular septum: a case report.

BACKGROUND: Human hydatid disease occurs after infection with Echinococcus granulosus, mainly involves liver and lung, while hydatid involves heart is infrequent. A great majority of hydatid diseases could be asymptomatic, and incidentally found through examination. Here, we reported a woman who suffered an isolated cardiac hydatid cyst located at the interventricular septum. CASE PRESENTATION: A 48-year-old woman presented intermittent chest pain was admitted to the hospital. Imaging examination revealed a cyst located at the interventricular septum near the right ventricular apex. Considering medical history, radiological findings and serological results, cardiac hydatid disease was suspected. The cyst was successfully removed, while pathological biopsy confirmed the diagnosis of infection of Echinococcus granulosus. Postoperative course was uneventful, the patient was discharged from hospital without complications. CONCLUSION: For symptomatic cardiac hydatid cyst, surgical resection is necessary to avoid progression of disease. During surgical procedure, appropriate methods to reduce the potential risk of hydatid cyst metastasis are essential. Besides surgery, combined with regular drug therapy is an effective strategy to prevent reappearance.

Protoscolicidal effects of curcumin nanoemulsion against protoscoleces of Echinococcus granulosus.

BACKGROUND: The aim of the present study was to assess in vitro protoscolicidal effects of curcumin nanoemulsion (CUR-NE) against protoscoleces of cystic echinococcosis (CE)/hydatid cysts. METHODS: The CUR-NE was prepared via spontaneous emulsification of soybean as the oil phase, a mixture of Tween 80 and Tween 85 as the surfactant, ethanol as the co-surfactant and distilled water. Various concentrations of CUR-NE (156, 312, 625 and 1250 µg/ml) were exposed to collected protoscoleces of infected sheep liver hydatid cysts for 10, 20, 30, 60 and 120 min. Viability of the protoscoleces were assessed using eosin exclusion test. Morphological changes of the protoscoleces were observed using differential interference contrast (DIC) microscopy. RESULTS: The mean particle size and zeta potential of CUR-NE included 60.4 ± 14.8 nm and - 16.1 ± 1.1 mV, respectively. Results showed that the viability of the protoscoleces decreased significantly with increases in CUR-NE concentrations (p < 0.001). The mortality rates of protoscoleces with exposure to concentrations of 1250 and 625 µg/ml of CUR-NE for 60 min were 94 and 73.33%, respectively. Mortality of the protoscoleces was 100% after 120 min of exposure to 1250 and 625 µg/ml concentrations of CUR-NE. Using NIC microscopy, extensively altered tegumental surface protoscoleces was observed after protoscoleces exposure to CUR-NE. CONCLUSION: The findings of the present study revealed the in vitro protoscolicidal potential of CUR-NE. Therefore, CUR-NEs are addressed as novel protoscolicidal agents, which can be used as an alternative natural medicine to kill the protoscoleces, owing to their low toxicity and significant inhibition potency. However, further studies are necessary to investigate pharmacologic and pharmacokinetics of CUR-NEs.

Antigen discovery by bioinformatics analysis and peptide microarray for the diagnosis of cystic echinococcosis.

BACKGROUND: Cystic echinococcosis (CE), caused by Echinococcus granulosus sensu lato, is a neglected zoonosis. Its diagnosis relies on imaging, supported by serology, while only imaging is useful for staging and follow-up. Since diagnostic tools and expertise are not widely available, new accurate and easily implementable assays for the diagnosis and follow-up of CE are highly needed. METHODOLOGY/PRINCIPAL FINDINGS: We aimed to identify new E. granulosus antigens through a bioinformatics selection applied to the parasite genome, followed by peptide microarray screening and validation in ELISA, using independent panels of sera from patients with hepatic CE and clinically relevant controls. From 950 proteins selected in silico, 2,379 peptides were evaluated by microarray for IgG reactivity and eight candidates selected for validation. Reactivity to one peptide was significantly higher in the CE group (p = 0.044), but had suboptimal diagnostic accuracy. CONCLUSIONS/SIGNIFICANCE: Here we performed bioinformatics analysis and peptide microarray for antigen discovery, useful for the diagnosis of CE. Eight candidates were selected and validated. Reactivity to one peptide associated to CE but had suboptimal diagnostic accuracy. Importantly, the database developed in this study may be used to identify other antigenic candidates for CE diagnosis and follow-up.

Single-Cell RNA Sequencing Reveals Unique Alterations in the Immune Panorama and Treg Subpopulations in Mice during the Late Stages of Echinococcus granulosus Infection.

Cystic echinococcosis (CE) is a common zoonotic parasitic disease that seriously impacts public health. However, the full spectrum of immune cell changes in Echinococcus granulosus infection, especially the negative immune regulation of subpopulations of regulatory T (Treg) cells, are not yet well understood. In this study, we used single-cell RNA sequencing and immunome repertoire (IR) sequencing to analyze 53,298 cells from the spleens and peripheral blood mononuclear cells (PBMCs) of healthy and E. granulosus-infected mice. We used immunofluorescence combined with RNA fluorescence in situ hybridization and quantitative real-time PCR to verify the sequencing results. Our results showed tissue-specific immune system alterations in mice infected with E. granulosus. E. granulosus-infected mice induced a subpopulation of CD4+ cells with type I interferon production potential. Furthermore, there were six different Treg cell subpopulations in vivo at three stages of differentiation, and Treg subpopulations of different classes and different stages of differentiation showed tissue specificity. After infection, the Lag3hi Treg and Gpr83+Igfbp4+ naive Treg subpopulations were specifically induced in PBMCs and the spleen, respectively. Furthermore, T follicular helper 2 (Tfh2) cells with high expression of Cxxc5 and Spock2 were found in E. granulosus-infected mice. Our data uncovered changes in the full spectrum of immune cells in mice following the late stages of E. granulosus infection, including subpopulations of cells that have not been emphasized in previous studies. These results further enrich the study of the bidirectional immunomodulatory mechanism and offer a different perspective for subsequent studies of infection in E. granulosus.

NOTCH signaling inhibition after DAPT treatment exacerbates alveolar echinococcosis hepatic fibrosis by blocking M1 and enhancing M2 polarization.

Alveolar echinococcosis (AE) is a lethal helminthic liver disease caused by persistent infection with Echinococcus multilocularis (E. multilocularis). Although more and more attention has been paid to the macrophages in E. multilocularis infection, the mechanism of macrophage polarization, a critical player in liver immunity, is seldom studied. NOTCH signaling is involved in cell survival and macrophage-mediated inflammation, but the role of NOTCH signaling in AE has been equally elusive. In this study, liver tissue samples from AE patients were collected and an E. multilocularis infected mouse model with or without blocking NOTCH signaling was established to analyze the NOTCH signaling, fibrotic and inflammatory response of the liver after E. multilocularis infection. Changes in polarization and origin of hepatic macrophages were analyzed by flow cytometry. In vitro qRT-PCR and Western blot assays were performed to analyze key receptors and ligands in NOTCH signaling. Our data demonstrated that hepatic fibrosis develops after AE, and the overall blockade of NOTCH signaling caused by DAPT treatment exacerbates the levels of hepatic fibrosis and alters the polarization and origin of hepatic macrophages. Blocking NOTCH signaling in macrophages after E. multilocularis infection downregulates M1 and upregulates M2 expression. The downregulation of NTCH3 and DLL-3 in the NOTCH signaling pathway is significant. Therefore, NOTCH3/DLL3 may be the key pathway in NOTCH signaling regulating macrophage polarization affecting fibrosis caused by AE.

Cystic Echinococcosis in Northern New Hampshire, USA.

In April 2022 and December 2022, the New Hampshire Department of Health and Human Services confirmed 2 cases of locally acquired human pulmonary cystic echinococcosis caused by Echinococcus granulosus tapeworms. Both patients reported dressing locally hunted moose and exposure to dogs.

Can Echinococcus Granulosus Infestation Prevent Pancreatic Cancer? An invivo Experimental Study.

Hydatid cyst is a zoonotic infestation caused by Echinococcus granulosus, and it is known that some parasites found in humans cause cancer in humans or some may have a protective effect against cancer. Cancer is one of the most serious health problems of today and it has been shown in some studies that parasites such as Echinococcus granulosus can have an inhibitory effect. The aim of this study was determined as whether Echinococcus granulosus has an inhibitory effect on exocrine pancreatic cancer with the help of the azaserine-rat model used in different cancer studies.  Material and Methods: During experimental process a total of 45 male Wistar rats used, 14-day-old male Wistar rats were divided into groups according to the experimental protocol, administered azaserine injection protocol or kept as a control group without azaserine injection. Animals are grouped as Group 1, Control Group (group not treated with Azaserine and not injected with protoscolex.) (E-A-) (n=7); Group 2, Group injected with (IP) Azaserine only (30mg/kg) (E-A+)  (n=8);Group 3, Group injected (IP) with protoscolex suspension of 1 cc only (E+A-) (n=15);Group 4, Group injected both Azaserine (IP) and protoscolex suspension (IP) (E+A+) (n=15). Atypical Acinar Cell Foci (AACF) load in the exocrine pancreas of each rat was measured quantitatively with the help of a video image analyzer and the AACF load was calculated with the help of a mathematical model. Results: Findings showed that the Atypical Acinar Cell Foci (AACF) burden was statistically significantly lower in the Azaserine+ protoscolex (Azaserine-injected-protoscolex-implanted) rat group compared to the other groups, suggesting that Echinococcosis in the azaserine-rat model could inhibit the development of precursor foci of neoplastic changes in the exocrine pancreas. Conclusion: The most significant aspect of our study is that it contributes new insights into the controversy that Echinococcosis suppresses pancreatic cancer.

Occurrence and economic impact of cystic echinococcosis in cattle slaughtered in the Matabeleland Region, Zimbabwe.

BACKGROUND: Cystic echinococcosis is a neglected disease responsible for millions of infections in both animals and humans. The toll on the global economy is estimated to be in the billions of United States dollars. Although much effort has been made by public health authorities to curb the rise in new infections, the occurrence of cystic echinococcosis is still being noted, especially in low-income countries. This study was conducted in the Matabeleland region of Zimbabwe to determine the incidence of cystic echinococcosis in bovines. METHODS: Meat inspection records from 2011 through 2021 kept at licensed abattoirs in the Matabeleland region were used to generate annual total figures of bovines slaughtered in the region, together with the corresponding number of organs condemned due to cystic echinococcosis. Descriptive statistics showing the overall incidence in each year, incidence per district, and cysts in infected organs were expressed as a percentage of the total number of cattle slaughtered per category. RESULTS: Bulawayo had the highest occurrence of cystic echinococcosis (1.359%, 95% CI, 1.254-1.412), followed by Matabeleland South (0.914%, 95% CI, 0.886-0.929) and then Matabeleland North (0.848%, 95% CI 0.818-0.863) provinces. The Bulilima, Bulawayo, and Bubi districts had the highest occurrences of cystic echinococcosis at 1.749%, 1.358%, and 1.286%, respectively. The lung was the most frequently affected organ (n = 7155; at 0.854%; and 95% CI, 0.8334-0.874%, followed by the liver (0.053%; 95% CI, 0.048-0.058%). The total direct economic loss due to organ condemnation during the study period was US$ 24,812.43.

EgSeverin and Eg14-3-3zeta from Echinococcus granulosus are potential antigens for serological diagnosis of echinococcosis in dogs and sheep.

Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the metacestode larva of Echinococcus granulosus. In this study, two-dimensional gel electrophoresis (2-DE) coupled with immunoblot analysis revealed that E. granulosus severin and 14-3-3zeta proteins (named EgSeverin and Eg14-3-3zeta, respectively) might be two potential biomarkers for serological diagnosis of echinococcosis. The recombinant EgSeverin (rEgSeverin, 45 kDa) and Eg14-3-3zeta (rEg14-3-3zeta, 35 kDa) were administered subcutaneously to BALB/c mice to obtain polyclonal antibodies for immunofluorescence analyses (IFAs). And IFAs showed that both proteins were located on the surface of protoscoleces (PSCs). Western blotting showed that both proteins could react with sera from E. granulosus-infected sheep, dog, and mice. Indirect ELISAs (rEgSeverin- and rEg14-3-3zeta-iELISA) were developed, respectively, with sensitivities and specificities ranging from 83.33% to 100% and a coefficient of variation (CV %) of less than 10%. The rEgSeverin-iELISA showed cross-reaction with both E. granulosus and E. multilocularis, while the rEg14-3-3zeta-iELISA showed no cross-reaction with other sera except for the E. granulosus-infected ones. The field sheep sera from Xinjiang and Qinghai were analyzed using rEgSeverin-iELISA, rEg14-3-3zeta-iELISA, and a commercial kit respectively, and no significant differences were found among the three methods (p > 0.05). However, the CE positive rates in sheep sera from Qinghai were significantly higher than those from Xinjiang (p < 0.01). Overall, the results suggest that EgSeverin and Eg14-3-3zeta could be promising diagnostic antigens for E. granulosus infection.

Past and present of diagnosis of echinococcosis: A review (1999-2021).

The larval forms of taeniid cestodes belonging to the genus Echinococcus are the source of the zoonotic infection known as echinococcosis. Alveolar and cystic echinococcosis are caused by Echinococcus multilocularis and Echinococcus granulosus (s. s), respectively. It is endemic in several regions of the world. In this systematic review, we describe diagnosis, and the species (human, canids, livestock, and small rodents) affected by cystic (CE) and alveolar echinococcosis (AE). From 1999 to 2021, we searched the online directory through PubMed, SCOPUS, Web of Science, and google scholar. Among the 37,700 records found in the online databases, 187 publications met our eligibility requirements. The majority of investigations employed a range of diagnostic methods, such as ELISA, imaging, copro-PCR, necropsy or arecoline hydrobromide purgation, morphological cestode confirmation, and fecal sieving/flotation to detect and confirm Echinococcus infection. ELISA was the most commonly used method followed by PCR, and imaging. The research team retrieved data describing the incidence or assessment of the diagnostic test for E. multilocularis in humans (N = 99), canids (N = 63), small ruminants (N = 13), large ruminants (N = 3), camel (N = 2), pigs (N = 2) and small mammals (N = 5). This study was conducted to explore the diagnostic tools applied to detect echinococcosis in humans as well as animals in prevalent countries, and to report the characteristic of new diagnostic tests for disease surveillance. This systematic review revealed that ELISA (alone or in combination) was the most common method used for disease diagnosis and diagnostic efficacy and prevalence rate increased when recombinant antigens were used. It is highly recommended to use combination protcols such as serological with molecular and imaging technique to diagnose disease. Our study identified scarcity of data of reporting echinococcosis in humans/ animals in low-income or developing countries particularly central Asian countries. Study reports in small rodents indicate their role in disease dissemination but real situation in these host is not reflected due to limited number of studies. Even though echinococcosis affects both public health and the domestic animal sector, therefore, it is important to devise new and strengthen implementation of the existing monitoring, judging, and control measures in this estimate.

Can contrast-enhanced ultrasound differentiate the type of hepatic echinococcosis: cystic echinococcosis or alveolar echinococcosis?

BACKGROUND: Hepatic echinococcosis (HE) is a zoonotic disease caused by Echinococcus, and Echinococcus granulosus and E. multilocularis are the most common, causing cystic echinococcosis (CE) and alveolar echinococcosis (AE), respectively. Contrast-enhanced ultrasound (CEUS) is an imaging technique which has been recommended for identifying focal lesions in the liver. However, the effect of CEUS on the differentiation of hepatic echinococcosis type remains unclear. METHODS: Twenty-five patients with 46 HE lesions confirmed by histopathology in our hospital from December 2019 to May 2022 were reviewed by conventional ultrasound (US) and CEUS examinations, respectively. After US was completed, the CEUS study was performed. A bolus injection of 1.0-1.2 ml of a sulfur hexafluoride-filled microbubble contrast agent (SonoVue®) was administered. The images and clips of the lesions by US and CEUS were reviewed retrospectively. The lesions detected using US were evaluated including the location, size, morphology, margin, internal echogenicity and the internal Doppler signal. The lesions detected using CEUS were evaluated including the enhancement degree, enhancement pattern and enhancing boundary in different phases. The diagnoses of lesions by US or CEUS were respectively recorded. By taking the histopathology as the gold standard, the paired Chi-square test was performed with statistical software (IBM SPSS; IBM Corp., Armonk, NY, USA), and the results of differentiation of HE type by US and CEUS were statistically analyzed. RESULTS: A total of 46 lesions were involved in 25 patients, including 10 males (40.0%) and 15 females (60.0%) aged 15-55 (42.9 ± 10.3) years. By histopathology, 24 lesions of nine patients were diagnosed as CE and 22 lesions of 16 patients were diagnosed as AE. Among the 46 HE lesions, compared with histopathological examination, the accuracy rate was 65.2% and 91.3% in US and CEUS findings, respectively. Among the 24 CE lesions, 13 lesions were correctly differentiated by US, and 23 by CEUS. The difference between US and CEUS was statistically significant (Chi-square test, [Formula: see text] = 8.10, df = 23, P < 0.005). Among the total 46 HE lesions, 30 lesions were correctly differentiated by US, and 42 by CEUS. The difference between US and CEUS was statistically significant (Chi-square test, [Formula: see text] = 10.08, df = 45, P < 0.005). CONCLUSIONS: CEUS is a more effective technique than US for differentiating the type of HE between CE and AE. It could be a reliable tool in the differentiation of HE.

Monoclonal antibody-based localization of major diagnostic antigens in metacestode tissue, excretory/secretory products, and extracellular vesicles of Echinococcus species.

Alveolar (AE) and cystic echinococcosis (CE) are severe parasitic zoonoses caused by the larval stages of Echinococcus multilocularis and E. granulosus sensu lato, respectively. A panel of 7 monoclonal antibodies (mAbs) was selected against major diagnostic epitopes of both species. The binding capacity of the mAbs to Echinococcus spp. excretory/secretory products (ESP) was analyzed by sandwich-ELISA, where mAb Em2G11 and mAb EmG3 detected in vitro extravesicular ESP of both E. multilocularis and E. granulosus s.s. These findings were subsequently confirmed by the detection of circulating ESP in a subset of serum samples from infected hosts including humans. Extracellular vesicles (EVs) were purified, and the binding to mAbs was analyzed by sandwich-ELISA. Transmission electron microscopy (TEM) was used to confirm the binding of mAb EmG3 to EVs from intravesicular fluid of Echinococcus spp. vesicles. The specificity of the mAbs in ELISA corresponded to the immunohistochemical staining (IHC-S) patterns performed on human AE and CE liver sections. Antigenic small particles designated as ''spems'' for E. multilocularis and ''spegs'' for E. granulosus s.l. were stained by the mAb EmG3IgM, mAb EmG3IgG1, mAb AgB, and mAb 2B2, while mAb Em2G11 reacted with spems and mAb Eg2 with spegs only. The laminated layer (LL) of both species was strongly visualized by using mAb EmG3IgM, mAb EmG3IgG1, mAb AgB, and mAb 2B2. The LL was specifically stained by mAb Em2G11 in E. multilocularis and by mAb Eg2 in E. granulosus s.l. In the germinal layer (GL), including the protoscoleces, a wide staining pattern with all structures of both species was observed with mAb EmG3IgG1, mAb EmG3IgM, mAb AgB, mAb 2B2, and mAb Em18. In the GL and protoscoleces, the mAb Eg2 displayed a strong E. granulosus s.l. specific binding, while mAb Em2G11 exhibited a weak granular E. multilocularis specific reaction. The most notable staining pattern in IHC-S was found with mAb Em18, which solely bound to the GL and protoscoleces of Echinococcus species and potentially to primary cells. To conclude, mAbs represent valuable tools for the visualization of major antigens in the most important Echinococcus species, as well as providing insights into parasite-host interactions and pathogenesis.