Echinococcus granulosus sensu stricto G1 is the predominant genotype in human and livestock isolates from Turkey and Iran, based on mitochondrial nad5 gene differentiation

Abstract Background Different genotypes of Echinococcus granulosus sensu stricto (s.s.) isolated from livestock and humans have been identified based on cox1 and nad1 genomic fragments. The present study was performed to differentiate the G1/G3 genotypes of Echinococcus granulosus (s.s.) isolated from humans and livestock (sheep and cattle) from Azerbaijan in northwestern Iran, Fars Province in southern Iran, and Van province in Eastern Turkey, using the nad5 gene fragment as a suitable marker to distinguish these two genotypes. Methods A total of 60 pathologically confirmed human hydatid cysts and 90 hydatid cyst samples from livestock were collected from Turkey and Iran. PCR was performed on all of the samples, targeting the nad5 gene. Based on PCR product quality, host type, and the geographical area where the samples were obtained, 36 of the samples were sequenced and were used in the phylogenetic analysis. Results Out of 36 evaluated samples, 26 (72.2%) samples belonged to G1, and 10 (27.8%) samples belonged to the G3 genotype. Out of 21 samples from Turkey, 16 (76.2%) were G1 and 5 (23.8%) were G3, while out of 15 samples from Iran, 10 (66.7%) were G1 and 5 (33.3%) were the G3 genotype. None of the samples isolated from humans in Iran or from sheep in Turkey were G3. Overall, between the two countries, 18.18% of E. granulosus isolates in cattle, 41.66% of isolates in sheep, and 23.07% of human samples were identified as G3, and the others as the G1 genotype. The G3 genotype was not detected in human samples from Iran or sheep samples from Turkey. Conclusion The findings of the study revealed that the G1 genotype of E. granulosus s.s. is the predominant genotype in humans and livestock, both in Turkey and Iran. The ratio of the E. granulosus s.s. G1 to G3 genotype was 3.2 in Turkey and 2 in Iran. The study also further confirmed that the nad5 gene properly differentiated the G1/G3 isolates of E. granulosus from both humans and livestock. Graphical 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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Molecular characterization of Echinococcus granulosus sensu stricto isolated from livestock of Qazvin, Iran

Infectious Disorders - Drug Targets ◽

10.2174/1871526520666201029130248 ◽

2020 ◽

Vol 20 ◽

Author(s):

Mojtaba Shahnazi ◽

Maryam Habibvand ◽

Morteza Ghanbari Johkool ◽

Elham Hajialilo ◽

Meysam Sharifdini ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Haplotype Diversity ◽

Sensu Stricto ◽

Cox1 Gene ◽

Intermediate Hosts ◽

Study Region ◽

Predominant Genotype ◽

Mitochondrial Cox1 ◽

Significant Difference

Introduction: Hydatidiosis is a serious parasitic disease in humans and livestock, worldwide. Echinococcus granulosus shows notable genetic variation among intermediate hosts. Several genotypes of the worm have been reported from different parts of Iran, but no information over the parasite genotypes status in the study region is available. The current study investigated the presence of different genotypes of E. granulosus in the livestock of Qazvin, Iran, by sequencing the mitochondrial Cox1 genes. Methodology: One hundred twenty E. granulosus isolates, including 30 from goats, 40 from cattle and 50 from sheep were collected from the slaughterhouses in Qazvin province. Mitochondrial Cox1 gene region was amplified by PCR and 30 isolates were sequenced. Phylogenetic analysis was done by using the MEGA 7.0 software. Morphological analysis was performed on rostellar hook length of protoscoleces. Result: All isolates were identified as E. granulosus sensu stricto (G1-G3 complex) among 17% of isolates clarified as G3 genotypes. G1 was the predominant genotype among the specimens. No significant difference between the rostellar hooks measurements of different genotypes was observed. Conclusion: Our findings confirmed the presence of E. granulosus sensu stricto in the region although further studies are required to determine the haplotype diversity of E. granulosus using different mitochondrial and nuclear genes.

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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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Molecular Study of Echinococcus granulosus Cestodes in Ukraine and the First Genetic Identification of Echinococcus granulosus Sensu Stricto (G1 Genotype) in the Country

Acta Parasitologica ◽

10.1007/s11686-021-00450-z ◽

2021 ◽

Author(s):

Viliam Šnábel ◽

Tetiana A. Kuzmina ◽

Anatoliy A. Antipov ◽

Oleksandr M. Yemets ◽

Serena Cavallero ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Molecular Study ◽

Sensu Stricto ◽

Genetic Identification ◽

Echinococcus Granulosus Sensu Stricto

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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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The optimum cut-off value to differentiate Echinococcus granulosus sensu stricto from other species of E. granulosus sensu lato using larval rostellar hook morphometry

Journal of Helminthology ◽

10.1017/s0022149x13000473 ◽

2013 ◽

Vol 89 (1) ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

S.V. Soriano ◽

N.B. Pierangeli ◽

L.A. Pianciola ◽

M. Mazzeo ◽

L.E. Lazzarini ◽

...

Keyword(s):

Molecular Epidemiology ◽

Echinococcus Granulosus ◽

Cystic Echinococcosis ◽

Control Strategies ◽

High Sensitivity ◽

Field Studies ◽

Sensu Stricto ◽

Roc Curve Analysis ◽

First Time

AbstractCystic echinococcosis caused by Echinococcus granulosus sensu lato is one of the most important helminth zoonoses in the world; it affects both humans and livestock. The disease is endemic in Argentina and highly endemic in the province of Neuquén. Considerable genetic and phenotypic variation has been demonstrated in E. granulosus, and ten different genotypes (G1–G10) have been identified using molecular tools. Echinococcus granulosus sensu lato may be considered a species complex, comprised of E. granulosus sensu stricto (G1–G3), E. equinus (G4), E. ortleppi (G5) and E. canadensis (G6–G10). In endemic areas, the characterization of cystic echinococcosis molecular epidemiology is important in order to apply adequate control strategies. A cut-off value for larval large hook total length to distinguish E. granulosus sensu stricto isolates from those produced by other species of the complex was defined for the first time. Overall, 1780 larval hooks of 36 isolates obtained from sheep (n= 11, G1), goats (n= 10, G6), cattle (n= 5, G6) and pigs (n= 10, G7) were analysed. Validation against molecular genotyping as gold standard was carried out using the receiver operating characteristic (ROC) curve analysis. The optimum cut-off value was defined as 26.5 μm. The proposed method showed high sensitivity (97.8%) and specificity (91.1%). Since in most endemic regions the molecular epidemiology of echinococcosis includes the coexistence of the widely distributed E. granulosus sensu stricto G1 strain and other species of the complex, this technique could be useful as a quick and economical tool for epidemiological and surveillance field studies, when fertile cysts are present.

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