Immunology of a unique biological structure: the Echinococcus laminated layer

The larval stages of the cestode parasites belonging to the genus Echinococcus grow within internal organs of humans and a range of animal species. The resulting diseases, collectively termed echinococcoses, include major neglected tropical diseases of humans and livestock. Echinococcus larvae are outwardly protected by the laminated layer (LL), an acellular structure that is unique to this genus. The LL is based on a fibrillar meshwork made up of mucins, which are decorated by galactose-rich O-glycans. In addition, in the species cluster termed E. granulosus sensu lato, the LL features nano-deposits of the calcium salt of myo-inositol hexakisphosphate (Insp6). The main purpose of our article is to update the immunobiology of the LL. Major recent advances in this area are (i) the demonstration of LL "debris" at the infection site and draining lymph nodes, (ii) the characterization of the decoy activity of calcium Insp6 with respect to complement, (iii) the evidence that the LL mucin carbohydrates interact specifically with a lectin receptor expressed in Kupffer cells (Clec4F), and (iv) the characterization of what appear to be receptor-independent effects of LL particles on dendritic cells and macrophages. Much information is missing on the immunology of this intriguing structure: we discuss gaps in knowledge and propose possible avenues for research.

Equinococosis: actualización en terminologías utilizadas desde la biología a la terapéutica / Echinococcosis: update on terminologies used from biology to therapeutics

RESUMEN: El diagnóstico y tratamiento de las equinococosis requiere de la participación de especialistas multidisciplinarios,por lo que, estandarizar la terminología utilizada es fundamental. Para armonizar la terminología sobre bases científicas y lingüísticas sólidas, la Asociación Mundial de Equinococosis generó un proceso de consenso formal. Los principales logros de este proceso fueron: realizar una actualización de la nomenclatura actual de Echinococcus spp.; generar un acuerdo sobre nombres de enfermedades causadas por Echinococcus spp.: equinococosis quística (EQ), equinococosis alveolar (EA) y equinococosis neotropical (NE), y la exclusión de los demás nombres; ejecutar un acuerdo sobre el uso restringido del adjetivo "hidatido" para referirse al quiste y al líquido debido a E. granulosus sensu lato; y un acuerdo sobre una descripción estandarizada de las intervenciones quirúrgicas para EQ, de acuerdo con el marco de "Acceso, Apertura, Resección e Integridad del quiste" (AARI). Además, se enumeraron 95 términos "aprobados" y 60 "rechazados". Las recomendaciones proporcionadas serán aplicables a publicaciones científicas y comunicación con profesionales. El objetivo de este manuscrito fue generar un documento de estudio referente al uso de terminología actualizada en echinococcosis desde la biología a la terapéutica.

SUMMARY: The diagnosis and treatment of echinococcosis requires the participation of different multidisciplinary specialists Therefore, standardizing the terminology used is essential. To harmonize terminology on a sound scientific and linguistic basis, the World Echinococcosis Association generated a formal consensus process. The main achievements of this process were to update the current nomenclature of Echinococcus spp .; to generate an agreement on names of diseases caused by Echinococcus spp .: cystic echinococcosis (CE), alveolar echinococcosis (EA) and neotropical echinococcosis (NE), and the exclusion of other names; execute an agreement on the restricted use of the adjective "hydatid" to refer to the cyst and the fluid due to E. granulosus sensu lato; and an agreement on a standardized description of surgical interventions for CE, according to the framework of "Cyst Access, Opening, Resection and Integrity" (AARI). In addition, 95 "approved" and 60 "rejected" terms were listed. The recommendations provided will be applicable to scientific publications and communication with professionals. The aim of this manuscript was to generate a study document referring to echinococcosis from biology to therapy.

Intramuscular hydatid cyst in the lower extremity: report of three cases

Abstract Hydatid cysts are rarely detected in the muscle tissue, even in endemic countries. Echinococcus granulosis is the most common causative agent, whereas Echinococcus alveolaris is rare. Three patients, two females and one male, with primary echinococcosis of the skeletal muscles are described in this report. Mean patient age was 33.3 ± 14.6 years. The disease occurred as soft tissue masses in all three patients, and no hydatid foci were detected elsewhere in the patients. Skeletal muscle echinococcosis should be considered in the differential diagnosis of limb masses, especially in endemic countries.

In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts / Avaliação in vitro do efeito de Nigella sativa e Punica granatum em protoescólices de cistos hidáticos

Abstract Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

Resumo Os cistos hidáticos (equinococose cística, CE) são comumente encontrados no fígado e nos pulmões dos hospedeiros afetados. A abordagem do tratamento geralmente é cirúrgica, e algumas drogas são administradas em conjunto antes da cirurgia para matar protoscólices e evitar choque anafilático devido ao vazamento de fluido hidático no peritônio e diminuir as oportunidades de recorrência. O presente estudo foi avaliar a eficácia in-vitro do extrato hidroalcoólico de casca de Punica granatum e Nigella sativa, sobre os protoescólices de cistos hidáticos, que foram coletados dos pulmões de camelos infectados. Concentrações dos extratos com diferentes tempos de exposição foram utilizados e um ensaio de viabilidade foi aplicado para medir o efeito escolicida. A N. sativa apresentou sua maior eficácia escolicida nas concentrações de 100 mg/mL e 10 mg/mL após 30 e 60 min. O extrato de casca de P. granatum mostrou sua máxima eficácia escolicida na concentração de 100 mg/mL após 120 min. Todos os experimentos do presente estudo revelaram que os extratos de N. sativa e P. granatum tiveram efeito escolicida dependente da dose e do tempo nos protoescólices dos cistos hidáticos de camelo. Pode-se concluir que o extrato de N. sativa é mais potente que o extrato de casca de P. granatum em relação ao efeito escolicida, mas a eficácia de ambos os extratos foi de correlação significativa moderada com o tempo de exposição e as concentrações.

A systematic review and meta-analysis of the genetic characterization of human echinococcosis in Iran, an endemic country / 한국역학회지

Human echinococcosis is an infectious disease caused by tapeworms belonging to the species Echinococcus. This parasite has a worldwide distribution and is considered a neglected tropical disease by the World Health Organization. Due to the diversity of Echinococcus spp. hosts, as well as variation in geographical, climatic, and socio-ethnic conditions, the question of the strains or genotypes of Echinococcus spp. that are involved in human infections is important. The aim of this study was to provide a summary of the available data on genotypes of Echinococcus obtained from the Iranian population. Four international databases (PubMed, Scopus, Science Direct, and Web of Science) and 4 Persian databases (Magiran, Scientific Information Database, Iran Medex, and IranDoc) were searched for cross-sectional studies that reported the genotypes of Echinococcus spp. in human echinococcosis cases using molecular methods in Iran through July 2018. The Newcastle-Ottawa Scale was used to assess the quality of the selected studies. A total of 559 cases of human cystic echinococcosis were reported in the 21 included articles. The majority of cases belonged to genotype G1 (89.2%; 95% confidence interval [CI], 80.1 to 95.8), genotype G6 (8.2%; 95% CI, 2.8 to 15.9), and genotype G3 (2.3%; 95% CI, 1.1 to 3.9). Since genotype G1 of Echinococcus appears to be the most prevalent genotype affecting humans in Iran, disease control initiatives aimed at sheep intermediate hosts may be the most beneficial. In addition, educational programs and serological screening in individuals may help reduce the national impact of the disease.

A systematic review and meta-analysis of the genetic characterization of human echinococcosis in Iran, an endemic country / 한국역학회지

Human echinococcosis is an infectious disease caused by tapeworms belonging to the species Echinococcus. This parasite has a worldwide distribution and is considered a neglected tropical disease by the World Health Organization. Due to the diversity of Echinococcus spp. hosts, as well as variation in geographical, climatic, and socio-ethnic conditions, the question of the strains or genotypes of Echinococcus spp. that are involved in human infections is important. The aim of this study was to provide a summary of the available data on genotypes of Echinococcus obtained from the Iranian population. Four international databases (PubMed, Scopus, Science Direct, and Web of Science) and 4 Persian databases (Magiran, Scientific Information Database, Iran Medex, and IranDoc) were searched for cross-sectional studies that reported the genotypes of Echinococcus spp. in human echinococcosis cases using molecular methods in Iran through July 2018. The Newcastle-Ottawa Scale was used to assess the quality of the selected studies. A total of 559 cases of human cystic echinococcosis were reported in the 21 included articles. The majority of cases belonged to genotype G1 (89.2%; 95% confidence interval [CI], 80.1 to 95.8), genotype G6 (8.2%; 95% CI, 2.8 to 15.9), and genotype G3 (2.3%; 95% CI, 1.1 to 3.9). Since genotype G1 of Echinococcus appears to be the most prevalent genotype affecting humans in Iran, disease control initiatives aimed at sheep intermediate hosts may be the most beneficial. In addition, educational programs and serological screening in individuals may help reduce the national impact of the disease.

Proteomic Analysis on Exosomes Derived from Patients’ Sera Infected with Echinococcus granulosus

Cystic echinococcosis (CE), a zoonotic disease caused by Echinococcus granulosus at the larval stage, predominantly develops in the liver and lungs of intermediate hosts and eventually results in organ malfunction or even death. The interaction between E. granulosus and human body is incompletely understood. Exosomes are nanosized particles ubiquitously present in human body fluids. Exosomes carry biomolecules that facilitate communication between cells. To the best of our knowledge, the role of exosomes in patients with CE is not reported. Here, we isolated exosomes from the sera of patients with CE (CE-exo) and healthy donors and subjected them to liquid chromatography-tandem mass spectrometry analysis. Proteomic analysis identified 49 proteins specifically expressed in CE-exo, including 4 proteins of parasitic origin. The most valuable parasitic proteins included tubulin alpha-1C chain and histone H4. And 8 proteins were differentially regulated in CE-exo (fold change>1.5), as analyzed with bioinformatic methods such as annotation and functional enrichment analyses. These findings may improve our understanding about the interaction between E. granulosus and human body, and may contribute to the diagnosis and prevention of CE.

A Case of Alveolar Echinococcosis Occurring in the Hilar Bile Duct

Echinococcosis is a disease caused by the Echinococcus species that parasitizes in humans. Alveolar echinococcosis (AE) which is caused by Echinococcus multilocularis is harmful to humans. AE mainly occurs in the liver and can be transferred to retroperitoneal lymph nodes, lung, brain, bone, spleen and other organs through lymphatic and blood vessels. Cholangiocarcinoma can occur in the intrahepatic and extrahepatic bile ducts and is more common in the hilar. We reported a case of hilar bile duct alveolar echinococcosis which was originally misdiagnosed an cholangiocarcinoma.

Prevalence and Molecular Characterization of Echinococcus granulosus Sensu Stricto in Northern Xinjiang, China

Echinococcus granulosus is an important zoonotic parasite globally causing cystic echinococcosis (CE) in humans and animals. In this study, prevalence of CE and variation of cox1 gene sequence were analyzed with isolates E. granulosus collected from different areas in northern Xinjiang, China. The survey showed that 3.5% of sheep and 4.1% of cattle were infected with CE. Fragment of cox1 was amplified from all the positive sheep and cattle samples by PCR. In addition, 26 positive samples across the 4 areas were included. The isolates were all E. granulosus sensu stricto (s.s.) containing 15 haplotypes (Hap1-15), and clustered into 2 genotypes, G1 (90.1%, 91/101) and G3 (9.9%, 10/101). Hap1 was the most common haplotype (48.5%, 49/101). Hap9 were found in humans samples, indicating that sheep and cattle reservoir human CE. It is indicate that E. granulosus may impact on control of CE in livestock and humans in the region.

Genetic diversity of Echinococcus vogeli in the western Brazilian Amazon

Human polycystic echinococcosis is a parasitic infection caused by the larval stage of Echinococcus vogeli, which occurs in rural areas of Central and South America. Until now, little information on the genetic variability of E. vogeli is available. Here, 32 samples from human-excised E. vogeli cysts had a 396-bp sequence of the mitochondrial cytochrome oxidase I (COI) gene sequenced and compared to another 17 COI sequences representing nine Echinococcus species. A Bayesian COI tree revealed that all E. vogeli sequences formed a monophyletic and well-supported clade with an E. vogeli reference sequence. The occurrence of geographically restricted E. vogeli COI haplotypes suggests retention of ancestral polymorphisms with little migration in Acre, Brazil.

Evaluación de la prueba rápida VIRAPID® Hidatidosis / Evaluation of the VIRAPID® Hydatidosis rapid test / Avaliação do teste rápido VIRAPID® Hidatidose

El diagnóstico presuntivo de la equinococosis quística (EQ) se basa en estudios clínicos e imagenológicos, en particular aquellos que aplican ultrasonido, en datos epidemiológicos, y se confirma con la serología específica. En las zonas endémicas rurales, donde los procedimientos imagenológicos no siempre se encuentran disponibles, al igual que la serología convencional debido a la falta de laboratorios suficientemente equipados, las pruebas de diagnóstico rápido (TDRs) pueden ser una herramienta de gran utilidad. Se evaluó aquí la precisión diagnóstica de VIRAPID® Hidatidosis (Vircell, España), comparado con dos métodos comerciales usados frecuentemente en este laboratorio, para el diagnóstico de la Hidatidosis: HIDATEST (Laboratorio Lemos, Argentina) y ELISA IgG Echinococcus (Vircell, España). El análisis se realizó sobre 224 muestras de igual número de pacientes; 48 (21,4%) pertenecían a pacientes con quistes probablemente hidatídicos, mientras que 10 (4,5%) a pacientes con quistes no parasitarios. Se agregaron a ellos, 166 (74,1%) muestras que resultaron previamente negativas en la casuística de este laboratorio. Para los tres métodos antes mencionados, la concordancia entre los resultados obtenidos fue evaluada por el estadístico Kappa, donde en todos los casos pudo apreciarse un grado de acuerdo casi excelente. La sensibilidad y especificidad de VIRAPID fueron de 94,1%, 98,9%, ELISA 90,6%, 99,4% e HIDATEST 96,0%, 96,2%, respectivamente. Los TDRs para hidatidosis pueden ser útiles en aquellos entornos con escasos recursos para establecer la certeza diagnóstica. La prueba VIRAPID® Hidatidosis demostró tener un muy buen rendimiento, aunque sería interesante evaluar la sensibilidad del test en presencia de quistes inactivos, lo cual, se supone podría plantear problemas para el diagnóstico.

The presumptive diagnosis of cystic echinococcosis (EQ) is based on clinical and imaging studies, particularly those that apply Ultrasound, in epidemiological data, and is confirmed with specific serology. In rural endemic areas, where imaging procedures are not always available, as with conventional serology due to the lack of sufficiently equipped laboratories, rapid diagnostic tests (TDRs) can be a very useful tool. The diagnostic accuracy of VIRAPID® Hydatidosis (Vircell, Spain) was evaluated here, compared with two commercial methods frequently used in the Laboratory, for the diagnosis of Hydatidosis: HIDATEST (Laboratorio Lemos, Argentina) and ELISA IgG Echinococcus (Vircell, Spain). The analysis was performed on 224 samples from the same number of patients; 48 (21.44%) belonged to patients with probable hydatid cysts, while 10 (4.46%) belonged to patients with non-parasitic cysts. A total of 166 (74.10%) samples that were previously negative in the casuistry of our Laboratory were added. For the three methods mentioned above, the agreement between the results obtained was evaluated by the Kappa statistics, where in all the cases an almost excellent degree of agreement could be appreciated. The Sensitivity and Specificity of VIRAPID® was 94.12%, 98.88%, ELISA 90.57%, 99.44% and HIDATEST 96.00%, 96.17%, respectively. TDRs for hydatidosis may be useful in settings with limited resources to establish diagnostic certainty. The VIRAPID® test showed a very good performance, although it would be interesting to evaluate the sensitivity of the test in the presence of inactive cysts, which can pose problems for the diagnosis.

O diagnóstico presuntivo de equinococose cística (EQ) baseia em estudos clínicos e de imagem, em particular aqueles que aplicam ultrasom, em dados epidemiológicos, e é confirmado com a sorologia específica. Em áreas endêmicas rurais, onde os procedimentos de imagens nem sempre estão disponíveis, como acontece com a sorologia convencional devido à falta de laboratórios suficientemente equipamentados, os testes de diagnóstico rápido (TDRs) podem ser uma ferramenta de grande utilidade. A precisão do diagnóstico de VIRapid® hidatidose (Vircell, Espanha) foi avaliada aqui, em comparação com dois métodos comerciais frequentemente usados neste laboratório, para o diagnóstico de Hidatidose: HIDATEST (Laboratório Lemos, Argentina) e ELISA IgG Echinococcus (Vircell, Espanha). A análise foi realizada em 224 amostras do mesmo número de pacientes; 48 (21,4%) pertenciam a pacientes com cistos provavelmente hidatídicos, enquanto 10 (4,5%) pertenciam a pacientes com cistos não parasitários. Adicionaram-se a eles 166 (74,1%) amostras que resultaram previamente negativas na casuística desse Laboratório. Para os três métodos citados acima, a concordância entre os resultados obtidos foi avaliada pela estatística Kappa, onde em todos os casos um grau de concordância quase excelente pôde ser apreciado. Sensibilidade e especificidade de VIRapid® foi de 94,1%, 98,9%, ELISA 90,6%, 99,4% e HIDATEST, 96,0% e 92,6%, respectivamente. Os TDRs para hidatidose podem ser úteis em ambientes com recursos limitados para estabelecer a certeza diagnóstica. O teste VIRAPID® Hidatidose mostrou ter desempenho muito bom, apesar de que seria interessante avaliar a sensibilidade do teste em presença de cistos inativos, o que se supõe pode causar problemas para o diagnóstico.

Genetic Diversity of Echinococcus granulosus Genotype G1 in Xinjiang, Northwest of China

Cystic echinococcosis (CE) caused by E. granulosus is a serious helminthic zoonosis in humans, livestock and wildlife. Xinjiang is one of high endemic province for CE in China. A total of 55 sheep and cattle livers containing echinococcal cysts were collected from slaughterhouses in Changji and Yining City, northern region of Xinjiang. PCR was employed for cloning 2 gene fragments, 12S rRNA and CO1 for analysis of phylogenetic diversity of E. granulosus. The results showed that all the samples collected were identified as G1 genotype of E. granulosus. Interestingly, YL5 and CJ75 strains were the older branches compared to those strains from France, Argentina, Australia. CO1 gene fragment showed 20 new genotype haploids and 5 new genotype haplogroups (H1-H5) by the analysis of Network 5.0 software, and the YLY17 strain was identified as the most ancestral haplotype. The major haplotypes, such as CJ75 and YL5 strains, showed identical to the isolates from Middle East. The international and domestic trade of livestock might contribute to the dispersal of different haplotypes for E. granulosus evolution.

Hiperendemia de equinococosis y fertilidad quística en porcinos del valle interandino de Huancarama, Perú / Echinococcosis hyper-endemic and cystic fertility in pigs of the inter-andean valley of Huancarama, Peru

El objetivo de la investigación fue determinar el porcentaje de infección de equinococosis en porcinos, fertilidad quística y factores asociados. Estudio de corte transversal. La población estuvo conformada por 579 porcinos faenados en el matadero de Huancarama. Los quistes hidatídicos fueron detectados mediante evaluación post mórtem. La fertilidad se determinó midiendo el diámetro de los quistes y observando microscópicamente la membrana germinativa en busca de protoescólices. El porcentaje de infección de equinococosis quística fue de 76,7% (IC95%: 73,2-80,3). Los porcinos mayores a dos años tuvieron más oportunidad de presentar quistes (RP 1,03; IC95%:1,0-1,1; p<0,01), al igual que los provenientes del distrito de Andahuaylas (RP 1,2; IC95%: 1,1-1,4; p<0,01). Los quistes pulmonares presentaron menor porcentaje de infección que los hepáticos. Se examinaron 346 quistes hidatídicos de 334 porcinos, se encontró una fertilidad total de 42,8% (IC95%: 37,4-48,1).

The objective of the research was to determine the percentage of echinococcosis infection in swine, cystic fertility and associated factors. Cross-sectional study. The population was formed by 579 pigs slaughtered in the slaughterhouse of Huancarama. Hydatid cysts were detected by postmortem evaluation. Fertility was determined by measuring the diameter of the cysts and microscopically observing the germinative membrane in search of protoscoleces. The percentage of cystic echinococcosis infection was 76.7% (CI 95%: 73.2-80.3). Pigs older than two years were more likely to have cysts (RP 1.03; CI 95%: 1.0-1.1; (P < 0.01), as well as those coming from the Andahuaylas district (RP 1.2; CI 95%: 1.1-1.4; P < 0.01). Lung cysts showed a lower percentage of infection than hepatic ones. We examined 346 hydatid cysts of 334 pigs, a total fertility of 42.8% was found (CI 95%: 37.4-48.1).

Diversidade genética de Echinococcus vogeli na Amazônia Ocidental brasileira e padronização e validação de método imunodiagnóstico para a equinococose policística humana, baseado em antígenos recombinantes / Genetic diversity of Echinococcus vogeli in the Western Brazilian Amazon and standardization and validation of an immunodiagnostic method for human polycystic echinococcosis, based on recombinant antigens

Equinococose policística humana (EP) é uma infecção parasitária causada pelo estágio larval de Echinococcus vogeli, que ocorre em áreas rurais e silvestres da América Central e do Sul. No entanto, pouca informação sobre a variabilidade genética de E. vogeli está disponível. Neste estudo, 32 amostras de cistos de E. vogeli excisados do fígado ou do mesentério de pacientes humanos tiveram uma seqüência de 396-pb do gene mitocondrial da citocromo oxidase 1 (cox1) sequenciada e comparada com outras 17 sequências de cox1 representando 9 espécies de Echinococcus. Uma árvore Bayesiana revelou que todas as sequências de E. vogeli formaram um clado monofilético e bem suportado com uma sequência de referência de E. vogeli. A ocorrência de haplótipos de E. vogeli geograficamente restritos sugere retenção de polimorfismos ancestrais com pouca migração no Acre, Brasil. Além disso, dois antígenos recombinantes (rAgB8/1 e rAgB8/2), correspondentes a subunidades do antígeno B (AgB) de E. granulosus e previamente caracterizados como de valor diagnóstico para a equinococose cística (EC) foram validados para o diagnóstico da EP. Estes antígenos apresentaram bom desempenho em ELISA para a detecção de anticorpos (IgG totais) específicos em soros de pacientes com EP, sendo potenciais substitutos para preparações antigênicas brutas derivadas de fluido hidático ou AgB nativo purificado. (AU)

Practical Algorisms for PCR-RFLP-Based Genotyping of Echinococcus granulosus Sensu Lato

Echinococcus granulosus sensu lato (s.l.) is a causative agent of cystic echinococcosis or cystic hydatid disease in humans and domestic and wild animals. The disease is a serious health problem in countries associated with poverty and poor hygiene practices, particularly in livestock raising. We introduced a practical algorism for genotyping the parasite, which may be useful to many developing countries. To evaluate the efficiency of the algorism, we genotyped 3 unknown strains isolated from human patients. We found that unknowns 1 and 3 were included in G1, G2, and G3 genotypes group and unknown 2 was included in G4 genotype (Echinococcus equinus) according to the algorisms. We confirmed these results by sequencing the 3 unknown isolates cox1 and nad1 PCR products. In conclusion, these new algorisms are very fast genotype identification tools that are suitable for evaluating E. granulosus s.l. isolated from livestock or livestock holders, particularly in developing countries.