Cystic Echinococcosis: Clinical, Immunological, and Biomolecular Evaluation of Patients from Sardinia (Italy)

Cystic echinococcosis (CE), a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus sensu lato (s.l.), is a worldwide public health problem. Echinococcus granulosus sensu stricto (s.s.), associated with G1 and G3 genotypes, is endemic with high prevalence in the Mediterranean basin. The parasite’s life cycle comprises definitive hosts (canids) and intermediate hosts (ruminants) and can occasionally involve humans. The main aim of this research was to confirm the diagnosis of 13 patients suspected of CE who presented different complications and needed the surgical removal of the cysts. We also wanted to understand and clarify more the diagnosis of echinococcosis in humans. For this purpose, the patients first underwent cyst evaluation by ultrasound (US), immunological analysis, and then total pericystectomy, followed by parasitological, histopathological, and molecular biology examinations of the cysts. US stadiated one CE1, one CE2, eight CE3b, one CE4, and two CE5; immunology evidenced nine positives; histopathology confirmed 11 CE cysts, of which 8 fertile presenting protoscoleces were identified as E. granulosus s.s. by molecular biology, genotyped as three G1 and four G3 by neighbor-joining (NJ) phylogenetic tree. In conclusion, the results showed that 11 patients were affected by E. granulosus s.s. G1 orG3, and 2 cystic neoformations were of non-parasitic origin.

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Molecular Characterization of Echinococcus granulosus sensu lato from Humans in Slovenia

Pathogens ◽

10.3390/pathogens9070562 ◽

2020 ◽

Vol 9 (7) ◽

pp. 562

Author(s):

Barbara Šoba ◽

Špela Gašperšič ◽

Darja Keše ◽

Tadeja Kotar

Keyword(s):

Molecular Characterization ◽

Echinococcus Granulosus ◽

Nadh Dehydrogenase ◽

Sensu Stricto ◽

Intermediate Hosts ◽

Western Balkans ◽

Nadh Dehydrogenase Subunit ◽

Important Intermediate ◽

Echinococcus Granulosus Sensu Lato

The larval form of tapeworms of the Echinococcus granulosus sensu lato species cluster cause an important zoonotic infection, cystic echinococcosis (CE). Molecular characterization of the cluster’s isolates from different hosts greatly contributes to a better understanding of its transmission dynamics. To date, no genetic information is available on CE in Slovenia. In this work, we characterized isolates from human CE cases. Parasite samples from 18 patients were collected, together with the patients’ demographic and clinical data. Genomic DNA was analyzed by conventional PCR and sequencing at four mitochondrial loci (cytochrome c oxidase subunit 1, cox1; NADH dehydrogenase subunit 1, nad1; NADH dehydrogenase subunit 5, nad5; and small ribosomal RNA, rrnS). Thirteen isolates were successfully amplified and sequenced. Seven (58.8%) patients were infected with E. granulosus sensu stricto (s.s.) G1, five (38.5%) with E. canadensis G7 and one (7.7%) with E. granulosus s.s. G3. Echinococcus canadensis G7, the pig genotype, was identified exclusively in autochthonous Slovenes, while the patients originating from the Western Balkans were all infected with E. granulosus s.s. Our findings suggest that pigs are important intermediate hosts for human CE in Slovenia.

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Species detection within the Echinococcus granulosus sensu lato complex by novel probe-based Real-Time PCRs

10.1101/2020.07.24.220756 ◽

2020 ◽

Author(s):

Pavlo Maksimov ◽

Hannes Bergmann ◽

Marion Wassermann ◽

Thomas Romig ◽

Bruno Gottstein ◽

...

Keyword(s):

Real Time ◽

Echinococcus Granulosus ◽

Cystic Echinococcosis ◽

Cost Effective ◽

Sensu Stricto ◽

Single Step ◽

Control Measures ◽

Chain Reactions ◽

Disease Epidemiology ◽

Echinococcus Granulosus Sensu Lato

AbstractInfections with eggs of Echinococcus granulosus sensu lato (s.l.) can cause cystic echinococcosis in intermediate host animals and humans. Upon ingestion of viable eggs, oncospheres hatch from the eggs and subsequently develop into fluid-filled larval cysts, most frequently in the liver or the lungs. The slowly growing cysts progressively interfere with organ function. The risk of infection is determined by the host range of the parasite, its pathogenicity and other epidemiologically relevant parameters, which differ significantly among the five species within the E. granulosus s.l. complex. It is therefore essential to diagnose the correct species within E. granulosus s.l. to help understand specific disease epidemiology and to facilitate effective implementation of control measures. For this purpose, simple, fast and cost-effective typing techniques are needed.We developed quantitative Real-Time Polymerase Chain Reactions (qPCRs) and corresponding sequence-specific hydrolysis DNA probes to target polymorphic regions in the mitochondrial genome of E. granulosus s.l.. In a single-step typing approach, we distinguished E. granulosus s.l. members in four epidemiologically relevant subgroups. These were E. granulosus sensu stricto (G1, G3), E. equinus (G4), E. ortleppi (G5) and the E. canadensis cluster (G6 to G8 and G10). The technique also allowed identification and differentiation of these species from other Echinococcus or Taenia taxa for samples isolated from cysts or faeces.Single-step genotyping techniques for the molecular diagnosis of Echinococcus spp. by qPCRs may not only improve diagnostic performance, but also our knowledge on the epidemiology of the parasites and help controlling the various agents of cystic echinococcosis.

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Cystic echinococcosis in a domestic cat (Felis catus) in Italy

Parasite ◽

10.1051/parasite/2018027 ◽

2018 ◽

Vol 25 ◽

pp. 25 ◽

Cited By ~ 4

Author(s):

Piero Bonelli ◽

Gabriella Masu ◽

Silvia Dei Giudici ◽

Davide Pintus ◽

Angela Peruzzu ◽

...

Keyword(s):

Cystic Echinococcosis ◽

Mitochondrial Gene ◽

Abdominal Distension ◽

Sensu Stricto ◽

Domestic Cat ◽

Intermediate Hosts ◽

Cytochrome Oxidase Subunit ◽

First Case ◽

Echinococcus Granulosus Sensu Lato ◽

Genotype G1

Echinococcus granulosus sensu lato is a zoonotic agent with a life cycle consisting of definitive hosts (dogs and wild carnivores), and intermediate hosts (usually ungulates). Other animals and humans may accidentally ingest eggs and contract cystic echinococcosis, acting as aberrant hosts. A 3-year-old neutered female cat was brought to a veterinary practice in Sassari (Italy) with abdominal distension. Ultrasound showed multiple intraperitoneal vesicles, which on laparotomy were found to be metacestodes of E. granulosus. Videos of the extraction of cysts are provided. Phylogenetic analysis based on a fragment of the cytochrome oxidase subunit 1 (cox1) mitochondrial gene identified the isolate as E. granulosus sensu stricto genotype G1, the most common genotype circulating in Europe and the Mediterranean basin. This is the first case report of cystic echinococcosis in domestic cats from Italy.

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Past and present of cystic echinococcosis in Bolivia

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009426 ◽

2021 ◽

Vol 15 (6) ◽

pp. e0009426

Author(s):

Viterman Ali ◽

Eddy Martinez ◽

Pamela Duran ◽

Erick Villena ◽

Peter Deplazes ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Cystic Echinococcosis ◽

Current Knowledge ◽

Epidemiological Studies ◽

Human Case ◽

Sensu Stricto ◽

Ultrasound Screening ◽

Intermediate Hosts ◽

La Paz ◽

Faecal Samples

Viable eggs of the canine intestinal tapeworm Echinococcus granulosus sensu lato (s.l.) infect various intermediate hosts causing cystic echinococcosis (CE). Furthermore, CE represents a serious zoonosis causing a significant global burden of disease. CE is highly endemic in South America, including Argentina, Brazil, Chile, Uruguay, and Peru. For Bolivia, no official data concerning the incidence in humans or the number of livestock and dogs infected are available. However, it is well known that CE occurs in Bolivia. We aim here to fill the gap in the current knowledge of the epidemiological situation of CE in Bolivia, providing a historical overview of documents published within the country, which have never been comprehensively reviewed. The very first documentation of E. granulosus infection in animals dates in 1910, while the first human case was reported in 1913. In total, 876 human CE cases have been reported in the scientific literature, with an apparent increase since the 1970s. In the absence of other epidemiological studies, the highest prevalence in human comes from Tupiza, Potosí Department, where 4.1% (51/1,268) of the population showed signs of CE at mass ultrasound screening in 2011. In the same report, 24% of dog faecal samples were positive for coproantigens of E. granulosus s.l. in ELISA. The highest prevalence in intermediate hosts reported at abattoir reached 37.5% in cattle from Potosí, followed by 26.9% in llamas from Oruro, 2.4% in pigs and 1.4% in sheep from La Paz. Finally, Echinococcus granulosus sensu stricto (s.s.), Echinococcus ortleppi (G5), and Echinococcus intermedius (G7) have been identified in Bolivia. Data reviewed here confirm that E. granulosus s.l. is circulating in Bolivia and that a proper prospective nationwide epidemiological study of CE is urgently needed to define transmission patterns as a basis for the planning and implementation of future control measurements.

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Echinococcus granulosus sensu stricto, Echinococcus ortleppi; and E. intermedius (G7) are present in Bolivia

Parasitology ◽

10.1017/s0031182020000529 ◽

2020 ◽

Vol 147 (9) ◽

pp. 949-956 ◽

Cited By ~ 2

Author(s):

V. Ali ◽

E. Martinez ◽

P. Duran ◽

M. A. Seláez ◽

M. Barragan ◽

...

Keyword(s):

Detailed Analysis ◽

Echinococcus Granulosus ◽

Cystic Echinococcosis ◽

Sensu Stricto ◽

Control Measures ◽

Santa Cruz ◽

La Paz ◽

Echinococcus Granulosus Sensu Stricto ◽

Faecal Samples ◽

Echinococcus Granulosus Sensu Lato

AbstractCystic 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.

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Phylogenetic Characteristics of Echinococcus granulosus Sensu Lato in Uzbekistan

The Korean Journal of Parasitology ◽

10.3347/kjp.2020.58.2.205 ◽

2020 ◽

Vol 58 (2) ◽

pp. 205-210 ◽

Cited By ~ 1

Author(s):

Hye-Jin Kim ◽

Tai-Soon Yong ◽

Myeong Heon Shin ◽

Kyu-Jae Lee ◽

Gab-Man Park ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Sensu Stricto ◽

Intermediate Hosts ◽

Livestock Farming ◽

Genetic Studies ◽

Human Sample ◽

The One ◽

First Time ◽

Dna Components ◽

Echinococcus Granulosus Sensu Lato

Echinococcosis occurs mainly in areas with heavy livestock farming, such as Central Asia, America, and Australia. Echinococcus granulosus sensu lato (s.l.) infection causes echinococcosis in intermediate hosts, such as sheep, cattle, goats, camels, and horses. Numerous cases of echinococcosis occur in Uzbekistan as stock farming is a primary industry. Epidemiological and genetic studies of E. granulosus s.l. are very important for mitigating its impact on public health and the economy; however, there are no such studies on E. granulosus s.l. in Uzbekistan. In the present study, to determine which genotypes exist and are transmitted, we isolated Echinococcus sp. from definitive hosts (one isolate each from jackal and dog) and intermediate hosts (52 isolates from humans and 6 isolates from sheep) in Uzbekistan and analyzed the isolates by sequencing 2 mitochondrial DNA components (cox1 and nad1). The results showed that all of isolates except one belonged to the E. granulosus sensu stricto (s.s.) G1 and G3 genotypes. Phylogenetic analysis based on cox1 sequences showed that 42 isolates from humans, 6 isolates from sheep, and one isolate from jackal were the G1 genotype, whereas the remaining 8 isolates from human and the one isolate from dog were the G3 genotype. These results suggest that the G1 and G3 genotypes of E. granulosus s.s. are predominant in Uzbekistan, and both wild animals and domestic animals are important for maintaining their life cycle. Only one isolate from human sample was confirmed to be E. eqiinus (G4 genotype), which is known to be for the first time.

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Response patterns in adventitial layer of Echinococcus granulosus sensu stricto cysts from naturally infected cattle and sheep

Veterinary Research ◽

10.1186/s13567-021-00936-8 ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Christian Hidalgo ◽

Caroll Stoore ◽

María Soledad Baquedano ◽

Ismael Pereira ◽

Carmen Franco ◽

...

Keyword(s):

Immune Response ◽

Echinococcus Granulosus ◽

Cystic Echinococcosis ◽

Direct Contact ◽

Sensu Stricto ◽

Response Patterns ◽

Granulomatous Reaction ◽

Adventitial Layer ◽

Laminated Layer ◽

Cattle And Sheep

AbstractCystic echinococcosis is a zoonotic disease caused by the metacestode of Echinococcus granulosus sensu lato. The disease is characterized by the development of cystic structures inside viscera of the intermediate host, mainly liver and lungs. These cysts are formed by three layers: germinal, laminated, and adventitial layer, the latter being the local host immune response. Metacestodes that develop protoscoleces, the infective stage to the definitive host, are termed fertile, whereas cysts that do not produce protoscoleces are termed non-fertile. Sheep usually harbor fertile cysts while cattle usually harbor non-fertile cysts. Adventitial layers with fibrotic resolution are associated to fertile cysts, whereas a granulomatous reaction is associated with non-fertile cysts. The aim of this study was to analyze cellular distribution in the adventitial layer of fertile and non-fertile E. granulosus sensu stricto cysts found in liver and lungs of cattle and sheep. A total of 418 cysts were analyzed, 203 from cattle (8 fertile and 195 non-fertile) and 215 from sheep (64 fertile and 151 non-fertile). Fertile cysts from cattle showed mixed patterns of response, with fibrotic resolution and presence of granulomatous response in direct contact with the laminated layer, while sheep fertile cysts always displayed fibrotic resolution next to the laminated layer. Cattle non-fertile cysts display a granulomatous reaction in direct contact with the laminated layer, whereas sheep non-fertile cysts display a granulomatous reaction, but in direct contact with the fibrotic resolution. This shows that cattle and sheep cystic echinococcosis cysts have distinct local immune response patterns, which are associated to metacestode fertility.

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Human Cystic Echinococcosis in Lebanon: A Retrospective Study and Molecular Epidemiology

Acta Parasitologica ◽

10.1007/s11686-021-00453-w ◽

2021 ◽

Author(s):

Gaelle Joanny ◽

Maria Grazia Cappai ◽

Francesca Nonnis ◽

Claudia Tamponi ◽

Giorgia Dessì ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Cystic Echinococcosis ◽

Epidemiological Data ◽

Sensu Stricto ◽

Control Measures ◽

Mediterranean Countries ◽

High Gene ◽

Human Infections ◽

And Control ◽

Human Cystic Echinococcosis

Abstract Purpose Human cystic echinococcosis (CE) is a zoonotic parasitic disease that constitutes a public health challenge and a socio-economic burden in endemic areas worldwide. No specific surveillance system of CE infections in humans exists in Lebanon. The incidence and trends over time have not been documented. The current study aimed to assess the demographic and epidemiologic features of human CE surgical cases over a 14-year period in the five main regions of Lebanon. Methods From 2005 to 2018, a total of 894 surgically confirmed cases of hydatidosis were recorded from five anatomy and pathology laboratories. Results The mean annual surgical incidence was 1.23/100,000 inhabitants. Over the span of these years, the incidence increased from 0.53 to 1.94 cases/100,000 inhabitants in 2005 and 2018, respectively. CE is present in Lebanon with an uneven distribution from one region to the other with higher prevalence in Bekaa (29.0%), a rural area where sheep raising is widespread. Human CE cases were more common in females (60.1%) than in males (39.9%) and a high burden of infection was reported for the age group of 30–39 years. Besides, 66.7% of the cases expressed only liver complications whereas, 20.5% showed predilection towards lungs. The 7.8% of cases presented cysts in other organs, and 1.3% showed multiple localizations. Additionally, predominant involvement of Echinococcus granulosus sensu stricto was recorded in human infections. Comparison of Echinococcus granulosus s.s. populations from different Mediterranean countries also revealed high gene flow among this region and sharing of alleles. Conclusion The current study is a step forward to fill the gap of knowledge for the hydatidosis in Lebanon where the lack of epidemiological data and control measures have resulted in higher incidence of human CE. Graphic Abstract

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Identification of Echinococcus granulosus Genotypes G1 and G3 by SNPs Genotyping Assays

Pathogens ◽

10.3390/pathogens10020125 ◽

2021 ◽

Vol 10 (2) ◽

pp. 125

Author(s):

Piero Bonelli ◽

Silvia Dei Giudici ◽

Angela Peruzzu ◽

Lorena Mura ◽

Cinzia Santucciu ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Animal Species ◽

Cost Effective ◽

Sensu Stricto ◽

Sybr Green ◽

Hydatid Cysts ◽

Genotype Identification ◽

Host Selectivity ◽

Nad5 Gene ◽

Echinococcus Granulosus Sensu Lato

Echinococcus granulosus sensu lato (s.l.) is the causative agent of cystic echinococcosis in animals and humans. Different E. granulosuss.l. genotypes exhibit great diversity in their life cycle, host selectivity and pathogenicity. For this reason, the study of genetic variation within Echinococcus species is of importance for their epidemiological implication. We employed two SNP genotyping technologies to distinguish G1 and G3 E. granulosus sensu stricto (s.s.). genotypes. The genotypes of DNA samples (n = 28) extracted from hydatid cysts of different animal species were identified by amplification and sequencing of a fragment of the mitochondrial nad5 gene. Two SYBR green and three TaqMan real time PCR assays were developed for targeting of three nad5 informative positions (SNP758, 1123, and 1380) known to be able to discriminate G1 from G3. Genotyping by SYBR Green PCR based on cycle threshold (Ct) with melting temperature (Tm) analysis and performed on SNP1123 and SNP1380 failed to identify one DNA sample. TaqMan assays for SNP758, 1123 and 1380 effectively confirmed genotype identification obtained by Sanger sequencing. Our results demonstrated that the combination of the three Taqman assays developed in this study represents a valuable and cost effective tool alternative to DNA sequencing for E. granulosus s.s. genotyping.

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Echinococcus granulosus sensu stricto: silencing of thioredoxin peroxidase impairs the differentiation of protoscoleces into metacestodes

Parasite ◽

10.1051/parasite/2018055 ◽

2018 ◽

Vol 25 ◽

pp. 57 ◽

Cited By ~ 5

Author(s):

Hui Wang ◽

Jun Li ◽

Chuanshan Zhang ◽

Baoping Guo ◽

Qin Wei ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Sensu Stricto ◽

Antioxidant Defence ◽

Culture Studies ◽

Antioxidant Defence System ◽

Thioredoxin Peroxidase ◽

Survival And Growth ◽

Echinococcus Granulosus Sensu Lato

Cystic echinococcosis (CE) is a cosmopolitan parasitic disease caused by infection with the larval stage of Echinococcus granulosus sensu lato. Thioredoxin peroxidase (TPx) may play an essential role in the antioxidant defence system of E. granulosus s.l. as neither catalase nor glutathione peroxidase activities have been detected in the parasite. However, it is not known whether TPx affects the survival and growth of E. granulosus s.l. during development. In this study, three fragments of siRNA specific for EgTPx (siRNA-1/2/3) were designed and transfected into protoscoleces of E. granulosus sensu stricto by electroporation. Quantitative real-time PCR and Western blotting analysis showed that siRNA-3 significantly reduced the expression of EgTPx. Coincidentally, knockdown of EgTPx expression in protoscoleces with siRNA-3 significantly reduced the viability of the parasite under oxidative stress induced by 0.6 mM H2O2. In vitro culture studies showed that protoscoleces treated with siRNA-3 reduced pre-microcyst formation. In vivo experiments showed that injecting mice intraperitoneally with protoscoleces treated with siRNA-3 resulted in a significant reduction in the number, size and weight of CE cysts compared with those of control animals. Silencing of EgTPx led to the impairment of growth of E. granulosus s.s. both in vitro and in vivo, indicating that EgTPx is an important factor for protoscoleces survival and plays an important role in the antioxidant defence against the host during development.

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