Molecular analysis reveals expansion of Fasciola hepatica distribution from Afghanistan to China

Thang, Tran Nhat; Hakim, Hakimullah; Rahimi, Razuin; Ichikawa‐Seki, Madoka

doi:10.1016/j.parint.2019.101930

Cited by 18 publications

(4 citation statements)

References 24 publications

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“…PEPCK was established as one of the reliable nuclear markers for species identification of Fasciola spp. (Thang et al, 2019), and it has been proposed as an egg antigen in S. mansoni (Asahi et al, 2000). Our group also identified PEPCK as an immunodominant antigen recognized by sera from F. hepatica infected patients (Marcilla et al, 2008).…”

Section: Discussionsupporting

confidence: 60%

Proteomic Analysis of Extracellular Vesicles From Fasciola hepatica Hatching Eggs and Juveniles in Culture

Trelis

Sánchez-López

Sánchez-Palencia

et al. 2022

Front. Cell. Infect. Microbiol.

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The identification of extracellular vesicles (EVs) in Fasciola hepatica has provided a new way to understand parasite-host communication. Most of the studies on EVs have focused on the adult stage of F. hepatica, but recently, the presence of EVs from different developmental stages has been reported. To better understand the potential role of EVs in the biology of the parasite and in the infection process, the protein cargo of EVs from embryonated eggs and newly-excysted juvenile (NEJs) flukes cultured up to 28 days, has been analyzed. EVs were isolated by size exclusion chromatography and evaluated by nanoparticle tracking analysis and transmission electron microscopy. LC-MS/MS proteomic analysis of EVs revealed the presence of 23 different proteins from embryonated egg-derived EVs and 29 different proteins from NEJ-derived EVs. Most of the identified proteins had been previously described in EVs from F. hepatica adults, including cytoskeletal proteins, glycolytic enzymes, stress-related proteins and tetraspanins. Nevertheless, EVs from hatching eggs and NEJs exhibited qualitative differences in composition, when compared to EVs form adults, including the absence of cathepsin cysteine peptidases. The differential content of the EVs released by the different developmental stages of the parasite reflect the intense activity of NEJs at this early stage, with several proteins involved in membrane traffic and cell physiology. This new set of identified proteins could help to understand key metabolic, biochemical and molecular mechanisms mediated by EVs that take place upon egg hatching and after parasite excystment.

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Section: Discussionsupporting

confidence: 60%

Proteomic Analysis of Extracellular Vesicles From Fasciola hepatica Hatching Eggs and Juveniles in Culture

Trelis

Sánchez-López

Sánchez-Palencia

et al. 2022

Front. Cell. Infect. Microbiol.

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“…Several previous reports attempted to identify the species of Fasciola flukes from these regions using morphological and morphometric features as well as conventional molecular markers (nuclear internal transcribed spacer 1 and 2) (Anh et al., 2018 ; Ichikawa & Itagaki, 2010 ; Ichikawa‐Seki, Ortiz, et al., 2016 ; Mizani et al., 2015 ; Teofanova et al., 2011 ); however, these findings have not been validated using more reliable nuclear gene markers. Recently, the nuclear genes, phosphoenol pyruvate carboxykinase (pepck) and DNA polymerase delta (pold), were found to be useful novel markers for the precise discrimination of F. hepatica, F. gigantica and hybrid Fasciola flukes without the need for gene sequencing that is expensive and not cost‐effective, particularly in the endemic countries (Hayashi et al., 2018 ; Hayashi, Ichikawa‐Seki, et al., 2016 ; Kasahara et al., 2021 ; Tang et al., 2016 ; Thang et al., 2019 ).…”

Section: Discussionmentioning

confidence: 99%

Morphological and molecular characterization of Fasciola isolates from livestock in Golestan province, northern Iran

Sharbatkhori

Nasibi

Mohammadi

et al. 2023

Veterinary Medicine & Sci

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Background: Fascioliasis, caused by the liver flukes Fasciola hepatica and Fasciola gigantica, is a global zoonotic helminthic disease. The livestock and human are the final hosts of the parasites. Northern Iran is an important endemic region for fascioliasis. Few studies have been conducted on the characterization of Fasciola isolates from eastern regions of the Caspian littoral of the country. Objective:The aim of the present study was to identify F. hepatica, F. gigantica and intermediate/hybrid forms of Fasciola isolates from livestock in Golestan province, northern Iran, using morphometric and molecular tools.Methods: Livestock livers naturally infected with Fasciola spp. were collected from Golestan slaughterhouse during 2019-2020. The worms were morphometrically studied using a calibrated stereomicroscope. Genomic DNA was extracted from all samples, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on internal transcribed spacer (ITS1) region using Rsa1 restriction enzyme. All the isolates were then analysed by multiplex PCR on Pepck region.Results: A total of 110 Fasciola isolates were collected from the infected livers, including 94 sheep, 12 cattle and 4 goats. Morphometric analysis of 61 adult Fasciola isolates indicated that, 44 and 17 isolates belonged to F. hepatica and F. gigantica, respectively.Eighty-one and 29 isolates belonged to F. hepatica and F. gigantica using ITS1-RFLP, respectively. However, Pepck Multiplex PCR indicated 72 F. hepatica, 26 F. gigantica and 12 intermediate/hybrid forms. All 12 hybrid isolates were found in sheep host. Two isolates were identified as F. gigantica using morphometry and F. hepatica using both molecular methods. Conclusion:The present study confirmed the existence of both F. hepatica and F. gigantica species and reported the first molecular evidence of hybrid Fasciola isolates in ruminants of Golestan province.

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“…A total of 1312 Fasciola flukes (470 F. hepatica, 609 F. gigantica, and 233 hybrid Fasciola) from 11 countries (Afghanistan, Algeria, Peru, Spain, Indonesia, Malaysia, Nigeria, Pakistan, Uganda, Japan, and Bangladesh) [8,9,11,12,[14][15][16][17][18][19][20] were used in the present study. Fragment analyses of nuclear pepck and pold and the nucleotide sequencing of mitochondrial nad1 have been performed in previous studies [8,9,11,12,[14][15][16][17][18][19][20]. Discrepancies between pepck and pold were observed among 7, 19, 6, 27, and 15 Fasciola isolates from Afghanistan, Algeria, Peru, Spain, and Nigeria, respectively.…”

Section: Fasciola Samplesmentioning

confidence: 99%

“…Although discrimination errors in the fragment pattern analysis of the multiplex PCR for pepck have been reported in F. hepatica isolates from Afghanistan [8], Algeria [9], Ecuador [10], and Spain [11], subsequent nucleotide sequencing of DNA fragment of pepck enabled precise species identification. Regarding pold, discrimination errors were observed in F. gigantica isolates from Nigeria [12].…”

Section: Introductionmentioning

confidence: 99%

Development of novel DNA marker for species discrimination of Fasciola flukes based on the fatty acid binding protein type I gene

et al. 2022

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Background Multiplex polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) for nuclear phosphoenolpyruvate carboxykinase (pepck) and polymerase delta (pold), respectively, have been used to differentiate Fasciola hepatica, F. gigantica, and hybrid Fasciola flukes. However, discrimination errors have been reported in both methods. This study aimed to develop a multiplex PCR based on a novel nuclear marker, the fatty acid binding protein type I (FABP) type I gene. Methods Nucleotide sequence variations of FABP type I were analyzed using DNA samples of F. hepatica, F. gigantica, and hybrid Fasciola flukes obtained from 11 countries in Europe, Latin America, Africa, and Asia. A common forward primer for F. hepatica and F. gigantica and two specific reverse primers for F. hepatica and F. gigantica were designed for multiplex PCR. Results Specific fragments of F. hepatica (290 bp) and F. gigantica (190 bp) were successfully amplified using multiplex PCR. However, the hybrid flukes contained fragments of both species. The multiplex PCR for FABP type I could precisely discriminate the 1312 Fasciola samples used in this study. Notably, no discrimination errors were observed with this novel method. Conclusions Multiplex PCR for FABP type I can be used as a species discrimination marker in place of pepck and pold. The robustness of the species-specific primer should be continuously examined using a larger number of Fasciola flukes worldwide in the future since nucleotide substitutions in the primer regions may cause amplification errors. Graphical abstract

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Molecular analysis reveals expansion of Fasciola hepatica distribution from Afghanistan to China

Cited by 18 publications

References 24 publications

Proteomic Analysis of Extracellular Vesicles From Fasciola hepatica Hatching Eggs and Juveniles in Culture

Proteomic Analysis of Extracellular Vesicles From Fasciola hepatica Hatching Eggs and Juveniles in Culture

Morphological and molecular characterization of Fasciola isolates from livestock in Golestan province, northern Iran

Development of novel DNA marker for species discrimination of Fasciola flukes based on the fatty acid binding protein type I gene

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