Fasciolopsis buski (Digenea: Fasciolidae) from China and India may represent distinct taxa based on mitochondrial and nuclear ribosomal DNA sequences

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Abstract

Background

Fasciolopsis buski is a zoonotic intestinal fluke infecting humans and pigs, but it has been seriously neglected. It is yet to know whether there is any genetic diversity among F. buski from different geographical locations, particularly in sequences of nuclear ribosomal DNA (rDNA) and mitochondrial (mt) DNA. Therefore, we determined the sequences of partial 18S, the complete internal transcribed spacer (ITS) rDNA and the complete mt genome of F. buski from China, compared the rDNA and mtDNA sequences with those of isolates from India and Vietnam, and assessed the phylogenetic relationships of this fluke and related fasciolid trematodes based on the mtDNA dataset.

Results

The complete mt genome sequence of F. buski from China is 14,833 bp, with 36 genes, including 12 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes ( rrnL and rrnS). The AT content of F. buski from China is 65.12%. The gene content and arrangement of the F. buski mt genome is similar to that of Fascioloides magna. Genetic distances between isolates of F. buski from China and India were high (28.2% in mtDNA, 13.2% in ITS-1 and 9.8% in ITS-2) and distinctly higher than the interspecific differences between Fasciola hepatica and Fasciola gigantica. The rDNA and mtDNA datasets for F. buski from China (isolate from pigs) and Vietnam (isolates from humans) were identical. The intergeneric differences in amino acid and nucleotide sequences among the genera Fasciolopsis, Fascioloides and Fasciola ranged between 24.64–25.56% and 26.35–28.46%, respectively.

Conclusions

Our results indicate that F. buski from China and India may represent distinct taxa, while F. buski in Vietnam and China represent the same species. These findings might have implications for the implementation of appropriate control strategies in different regions. Further studies are needed to decode mtDNA and rDNA sequences of F. buski from various geographical isolates for the better understanding of the species complex of F. buski.

Electronic supplementary material

The online version of this article (doi:10.1186/s13071-017-2039-2) contains supplementary material, which is available to authorized users.

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Most cited references 32

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tRNAscan-SE: A Program for Improved Detection of Transfer RNA Genes in Genomic Sequence

T. M. Lowe, S. Eddy (1997)

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Fascioliasis and other plant-borne trematode zoonoses.

S Mas-Coma, M D Bargues, M. A. Valero (2005)

Fascioliasis and other food-borne trematodiases are included in the list of important helminthiases with a great impact on human development. Six plant-borne trematode species have been found to affect humans: Fasciola hepatica, Fasciola gigantica and Fasciolopsis buski (Fasciolidae), Gastrodiscoides hominis (Gastrodiscidae), Watsonius watsoni and Fischoederius elongatus (Paramphistomidae). Whereas F. hepatica and F. gigantica are hepatic, the other four species are intestinal parasites. The fasciolids and the gastrodiscid cause important zoonoses distributed throughout many countries, while W. watsoni and F. elongatus have been only accidentally detected in humans. Present climate and global changes appear to increasingly affect snail-borne helminthiases, which are strongly dependent on environmental factors. Fascioliasis is a good example of an emerging/re-emerging parasitic disease in many countries as a consequence of many phenomena related to environmental changes as well as man-made modifications. The ability of F. hepatica to spread is related to its capacity to colonise and adapt to new hosts and environments, even at the extreme inhospitality of very high altitude. Moreover, the spread of F. hepatica from its original European range to other continents is related to the geographic expansion of its original European lymnaeid intermediate host species Galba truncatula, the American species Pseudosuccinea columella, and its adaptation to other lymnaeid species authochthonous in the newly colonised areas. Although fasciolopsiasis and gastrodiscoidiasis can be controlled along with other food-borne parasitoses, fasciolopsiasis still remains a public health problem in many endemic areas despite sustained WHO control programmes. Fasciolopsiasis has become a re-emerging infection in recent years and gastrodiscoidiasis, initially supposed to be restricted to Asian countries, is now being reported in African countries.

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Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation.

Robin Gasser, Min Shan Hu, Neil B Chilton … (2006)

The accurate analysis of genetic variation has major implications in many areas of biomedical research, including the identification of infectious agents (such as parasites), the diagnosis of infections, and the detection of unknown or known disease-causing mutations. Mutation scanning methods, including PCR-coupled single-strand conformation polymorphism (SSCP), have significant advantages over many other nucleic acid techniques for the accurate analysis of allelic and mutational sequence variation. The present protocol describes the SSCP method of analysis, including all steps from the small-scale isolation of genomic DNA and PCR amplification of target sequences, through to the gel-based separation of amplicons and scanning for mutations by SSCP (either by the analysis of radiolabeled amplicons in mutation detection enhancement (MDE) gels or by non-isotopic SSCP using precast GMA gels). The subsequent sequence analysis of polymorphic bands isolated from gels is also detailed. The SSCP protocol can readily detect point mutations for amplicon sizes of up to 450-500 bp, and usually takes 1-2 days to carry out. This user-friendly, low-cost, potentially high-throughput platform has demonstrated the utility to study a wide range of pathogens and diseases, and has the potential to be applied to any gene of any organism.

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Author and article information

Contributors

Jun Ma: dreamerjm@163.com

Miao-Miao Sun: smm0608@126.com

Jun-Jun He: hejunjun617@163.com

Guo-Hua Liu: liuguohua5202008@163.com

Lin Ai: 252120016@qq.com

Mu-Xin Chen: ipdchenmuxin@163.com

Xing-Quan Zhu: xingquanzhu1@hotmail.com

Journal

Journal ID (nlm-ta): Parasit Vectors

Journal ID (iso-abbrev): Parasit Vectors

Title: Parasites & Vectors

Publisher: BioMed Central (London )

ISSN (Electronic): 1756-3305

Publication date (Electronic): 22 February 2017

Publication date PMC-release: 22 February 2017

Publication date Collection: 2017

Volume: 10

Electronic Location Identifier: 101

Affiliations

[1 ]ISNI 0000 0001 0526 1937, GRID grid.410727.7, State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, , Chinese Academy of Agricultural Sciences, ; Lanzhou, Gansu Province 730046 People’s Republic of China

[2 ]ISNI 0000 0004 1760 4804, GRID grid.411389.6, College of Animal Science and Technology, , Anhui Agricultural University, ; Hefei, Anhui Province 230036 People’s Republic of China

[3 ]GRID grid.257160.7, , College of Veterinary Medicine, Hunan Agricultural University, ; Changsha, Hunan Province 410128 People’s Republic of China

[4 ]ISNI 0000 0000 8803 2373, GRID grid.198530.6, National Institute of Parasitic Diseases, , Chinese Center for Disease Control and Prevention, ; Shanghai, 200025 People’s Republic of China

[5 ]GRID grid.268415.c, Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, , Yangzhou University College of Veterinary Medicine, ; Yangzhou, Jiangsu Province 225009 People’s Republic of China

Article

Publisher ID: 2039

DOI: 10.1186/s13071-017-2039-2

PMC ID: 5322651

PubMed ID: 28228149

SO-VID: 37013baf-fdd1-4d14-b292-26659fab2eda

License:

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

History

Date received : 24 December 2016

Date accepted : 15 February 2017

Funding

Funded by: the Fundamental Research Funds of Chinese Academy of Agricultural Sciences

Award ID: Y2016JC05

Award Recipient : Xing-Quan Zhu

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ScienceOpen disciplines: Parasitology

Keywords: fasciolopsis buski,mitochondrial dna,nuclear ribosomal dna,phylogenetic analysis

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ScienceOpen disciplines: Parasitology

Keywords: fasciolopsis buski, mitochondrial dna, nuclear ribosomal dna, phylogenetic analysis

Fasciolopsis buski (Digenea: Fasciolidae) from China and India may represent distinct taxa based on mitochondrial and nuclear ribosomal DNA sequences

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Background

Results

Conclusions

Electronic supplementary material

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Malaria vaccine development

Most cited references 32

tRNAscan-SE: A Program for Improved Detection of Transfer RNA Genes in Genomic Sequence

Fascioliasis and other plant-borne trematode zoonoses.

Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation.

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