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The complete mitogenome of the Asian lung fluke Paragonimus skrjabini miyazakii and its implications for the family Paragonimidae (Trematoda: Platyhelminthes)

Published online by Cambridge University Press:  23 August 2022

Thanh Hoa Le*
Affiliation:
Immunology Department, Institute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), 18. Hoang Quoc Viet Rd., Cau Giay, Hanoi, Vietnam Graduate University of Science and Technology (GUST), Vietnam Academy of Science and Technology (VAST), 18. Hoang Quoc Viet Rd., Cau Giay, Hanoi, Vietnam
Khue Thi Nguyen
Affiliation:
Immunology Department, Institute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), 18. Hoang Quoc Viet Rd., Cau Giay, Hanoi, Vietnam
Linh Thi Khanh Pham
Affiliation:
Immunology Department, Institute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), 18. Hoang Quoc Viet Rd., Cau Giay, Hanoi, Vietnam
Huong Thi Thanh Doan
Affiliation:
Immunology Department, Institute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), 18. Hoang Quoc Viet Rd., Cau Giay, Hanoi, Vietnam Graduate University of Science and Technology (GUST), Vietnam Academy of Science and Technology (VAST), 18. Hoang Quoc Viet Rd., Cau Giay, Hanoi, Vietnam
Takeshi Agatsuma
Affiliation:
Department of Environmental Health Sciences, Kochi Medical School, Kohasu, Oko-cho 185-1, Nankoku, Kochi, 783-8505, Japan
David Blair*
Affiliation:
College of Marine and Environmental Sciences, James Cook University, Townsville, Queensland 4811, Australia
*
Author for correspondence: Thanh Hoa Le, E-mail: [email protected]; David Blair, E-mail: [email protected]
Author for correspondence: Thanh Hoa Le, E-mail: [email protected]; David Blair, E-mail: [email protected]

Abstract

The complete circular mitogenome of Paragonimus skrjabini miyazakii (Platyhelminthes: Paragonimidae) from Japan, obtained by PacBio long-read sequencing, was 17 591 bp and contained 12 protein-coding genes (PCGs), 2 mitoribosomal RNA and 22 transfer RNA genes. The atp8 gene was absent, and there was a 40 bp overlap between nad4L and nad4. The long non-coding region (4.3 kb) included distinct types of long and short repeat units. The pattern of base usage for PCGs and the mtDNA coding region overall in Asian and American Paragonimus species (P. s. miyazakii, P. heterotremus, P. ohirai and P. kellicotti) and the Indian form of P. westermani was T > G > A > C. On the other hand, East-Asian P. westermani used T > G > C > A. Five Asian and American Paragonimus species and P. westermani had TTT/Phe, TTG/Leu and GTT/Val as the most frequently used codons, whereas the least-used codons were different in each species and between regional forms of P. westermani. The phylogenetic tree reconstructed from a concatenated alignment of amino acids of 12 PCGs from 36 strains/26 species/5 families of trematodes confirmed that the Paragonimidae is monophyletic, with 100% nodal support. Paragonimus skrjabini miyazakii was resolved as a sister to P. heterotremus. The P. westermani clade was clearly separate from remaining congeners. The latter clade was comprised of 2 subclades, one of the East-Asian and the other of the Indian Type 1 samples. Additional mitogenomes in the Paragonimidae are needed for genomic characterization and are useful for diagnostics, identification and genetic/ phylogenetic/ epidemiological/ evolutionary studies of the Paragonimidae.

Type
Research Article
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

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References

References

Agatsuma, T, Ketudat, P, Thaithong, S, Shibahara, T, Sugiyama, H, Habe, S, Terasaki, K and Kawashima, K (1992) Electrophoretic analysis of a natural population of the Thai Paragonimus heterotremus and its genetic relationship to the three Japanese species P. miyazakii, P. ohirai and P. westermani. Parasitology Research 78, 463468.CrossRefGoogle Scholar
Benson, G (1999) Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Research 27, 573580.CrossRefGoogle ScholarPubMed
Bernt, M, Braband, A, Schierwater, B and Stadler, PF (2013 a) Genetic aspects of mitochondrial genome evolution. Molecular Phylogenetics and Evolution 69, 328338.CrossRefGoogle ScholarPubMed
Bernt, M, Donath, A, Jühling, F, Externbrink, F, Florentz, C, Fritzsch, G, Pütz, J, Middendorf, M and Stadler, PF (2013 b) MITOS: improved de novo metazoan mitochondrial genome annotation. Molecular Phylogenetics and Evolution 69, 313319.CrossRefGoogle ScholarPubMed
Biswal, DK, Chatterjee, A, Bhattacharya, A and Tandon, V (2014) The mitochondrial genome of Paragonimus westermani (Kerbert, 1878), the Indian isolate of the lung fluke representative of the family Paragonimidae (Trematoda). Peer J 2, e484.CrossRefGoogle Scholar
Blair, D (2008) Family Paragonimidae Dollfus, 1939. In Bray, RA, Gibson, DI and Jones, A (eds), Keys to the Trematoda, vol. 3. London: CAB International and Natural History Museum, pp. 271273.Google Scholar
Blair, D (2019) Paragonimiasis. In Toledo, R and Fried, B (eds), Digenetic Trematodes. Springer, 2nd Edn. Cham, Switzerland: Springer Nature Switzerland AG, pp. 105138.CrossRefGoogle Scholar
Blair, D (2022) Lung flukes of the genus Paragonimus: ancient and re-emerging pathogens. Parasitology 149, 12861295. https://doi.org/10.1017/S0031182022000300CrossRefGoogle ScholarPubMed
Blair, D, Agatsuma, T, Watanobe, T, Okamoto, M and Ito, A (1997) Geographical genetic structure within the human lung fluke, Paragonimus westermani, detected from DNA sequences. Parasitology 115, 411417.CrossRefGoogle ScholarPubMed
Blair, D, Chang, Z, Chen, M, Cui, A, Wu, B, Agatsuma, T, Iwagami, M, Corlis, D, Fu, C and Zhan, X (2005) Paragonimus skrjabini Chen, 1959 (Digenea: Paragonimidae) and related species in eastern Asia: a combined molecular and morphological approach to identification and taxonomy. Systematic Parasitology 60, 121.CrossRefGoogle ScholarPubMed
Blair, D, Nawa, Y, Mitreva, M and Doanh, PN (2016) Gene diversity and genetic variation in lung flukes (genus Paragonimus). Transactions of the Royal Society of Tropical Medicine and Hygiene 110, 612.CrossRefGoogle Scholar
Boore, JL (1999) Animal mitochondrial genomes. Nucleic Acids Research 27, 17671780.CrossRefGoogle ScholarPubMed
Cai, XQ, Liu, GH, Song, HQ, Wu, CY, Zou, FC, Yan, H, Yuan, ZG, Lin, RQ and Zhu, XQ (2012) Sequences and gene organization of the mitochondrial genomes of the liver flukes Opisthorchis viverrini and Clonorchis sinensis (Trematoda). Parasitology Research 110, 235243.CrossRefGoogle Scholar
Chai, JY and Jung, BK (2018) Paragonimus spp. In Rose, JB and Jiménez-Cisneros, B (eds), Global Water Pathogen Project, pp. http://www.waterpathogens.org. http://www.waterpathogens.org/book/paragonimus Michigan State University, E. Lansing, MI, UNESCO. pp 1–23. https://doi.org/10.14321/waterpathogens.46.CrossRefGoogle Scholar
Conant, GC and Wolfe, KH (2008) GenomeVx: simple web-based creation of editable circular chromosome maps. Bioinformatics (Oxford, England) 24, 861862.CrossRefGoogle ScholarPubMed
Devi, KR, Narain, K, Mahanta, J, Nirmolia, T, Blair, D, Saikia, SP and Agatsuma, T (2013) Presence of three distinct genotypes within the Paragonimus westermani complex in northeastern India. Parasitology 140, 7686.Google Scholar
Doanh, NP, Tu, AL, Bui, TD, Loan, TH, Nonaka, N, Horii, Y, Blair, D and Nawa, Y (2016) Molecular and morphological variation of Paragonimus westermani in Vietnam with records of new second intermediate crab hosts and a new locality in a northern province. Parasitology 143, 16391646.CrossRefGoogle Scholar
Gurevich, A, Saveliev, V, Vyahhi, N and Tesler, G (2013) QUAST: quality assessment tool for genome assemblies. Bioinformatics (Oxford, England) 29, 10721075.CrossRefGoogle ScholarPubMed
Hardman, M and Hardman, LM (2006) Comparison of the phylogenetic performance of neodermatan mitochondrial protein-coding genes. Zoologica Scripta 35, 655665.CrossRefGoogle Scholar
Kamo, H, Nishida, H, Hatsushika, R and Tomimura, T (1961) On the occurrence of a new lung fluke, Paragonimus miyazakii n.sp. in Japan. (Trematoda: Troglotrematidae). Yonago Acta Medica 5, 4352.Google Scholar
Katoh, K and Standley, DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30, 772780.CrossRefGoogle ScholarPubMed
Kinkar, L, Korhonen, PK, Cai, H, Gauci, CG, Lightowlers, MW, Saarma, U, Jenkins, DJ, Li, J, Li, J, Young, ND and Gasser, RB (2019) Long-read sequencing reveals a 4.4 kb tandem repeat region in the mitogenome of Echinococcus granulosus (sensu stricto) genotype G1. Parasites and Vectors 12, 238.CrossRefGoogle ScholarPubMed
Kinkar, L, Young, ND, Sohn, WM, Stroehlein, AJ, Korhonen, PK and Gasser, RB (2020) First record of a tandem-repeat region within the mitochondrial genome of Clonorchis sinensis using a long-read sequencing approach. PLOS Neglected Tropical Diseases 14, e0008552.CrossRefGoogle ScholarPubMed
Koren, S, Walenz, BP, Berlin, K, Miller, JR, Bergman, NH and Phillippy, AM (2017) Canu: scalable and accurate long-read assembly via adaptive k-mer weighting and repeat separation. Genome Research 27, 722736.CrossRefGoogle ScholarPubMed
Kumar, S, Stecher, G, Li, M, Knyaz, C and Tamura, K (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35, 15471549.CrossRefGoogle ScholarPubMed
Laslett, D and Canback, B (2008) ARWEN: a program to detect tRNA genes in metazoan mitochondrial nucleotide sequences. Bioinformatics (Oxford, England) 24, 172175.CrossRefGoogle ScholarPubMed
Le, TH, Blair, D and McManus, DP (2001) Complete DNA sequence and gene organization of the mitochondrial genome of the liverfluke, Fasciola hepatica L. (Platyhelminthes; Trematoda). Parasitology 123, 609621.CrossRefGoogle Scholar
Le, TH, Blair, D and McManus, DP (2002) Mitochondrial genomes of parasitic flatworms. Trends in Parasitology 18, 206213.CrossRefGoogle ScholarPubMed
Le, TH, Nguyen, KT, Nguyen, NTB, Doan, HTT, Agatsuma, T and Blair, D (2019) The complete mitochondrial genome of Paragonimus ohirai (Paragonimidae: Trematoda: Platyhelminthes) and its comparison with P. westermani congeners and other trematodes. PeerJ 7, e7031.Google Scholar
Le, TH, Pham, LTK, Doan, HTT, Le, XTK, Saijuntha, W, Rajapakse, RPVJ and Lawton, SP (2020) Comparative mitogenomics of the zoonotic parasite Echinostoma revolutum resolves taxonomic relationships within the ‘E. revolutum’ species group and the Echinostomata (Platyhelminthes: Digenea). Parasitology 147, 566576.CrossRefGoogle Scholar
Li, Y, Qiu, YY, Zeng, MH, Diao, PW, Chang, QC, Gao, Y, Zhang, Y and Wang, CR (2019) The complete mitochondrial genome of Echinostoma miyagawai: comparisons with closely related species and phylogenetic implications. Infection Genetics and Evolution 75, 10396.CrossRefGoogle ScholarPubMed
Liu, GH, Gasser, RB, Young, ND, Song, HQ, Ai, L and Zhu, XQ (2014) Complete mitochondrial genomes of the ‘intermediate form’ of Fasciola and Fasciola gigantica, and their comparison with F. hepatica. Parasites and Vectors 7, 150.CrossRefGoogle ScholarPubMed
Lowe, TM and Chan, PP (2016) tRNAscan on-line: search and contextual analysis of transfer RNA genes. Nucleic Acids Research 44, W54W57.CrossRefGoogle Scholar
Ma, J, He, JJ, Liu, GH, Leontovyč, R, Kašný, M and Zhu, XQ (2016) Complete mitochondrial genome of the giant liver fluke Fascioloides magna (Digenea: Fasciolidae) and its comparison with selected trematodes. Parasites and Vectors 9, 429.CrossRefGoogle ScholarPubMed
Oey, H, Zakrzewski, M, Gravermann, K, Young, ND, Korhonen, PK, Gobert, GN, Nawaratna, S, Hasan, S, Martínez, DM, You, H, Lavin, M, Jones, MK, Ragan, MA, Stoye, J, Oleaga, A, Emery, AM, Webster, BL, Rollinson, D, Gasser, RB, McManus, DP and Krause, L (2019 a) Whole-genome sequence of the bovine blood fluke Schistosoma bovis supports interspecific hybridization with S. haematobium. PLOS Pathogens 15, e1007513.CrossRefGoogle ScholarPubMed
Oey, H, Zakrzewski, M, Narain, K, Devi, KR, Agatsuma, T, Nawaratna, S, Gobert, GN, Jones, MK, Ragan, MA, McManus, DP and Krause, L (2019 b) Whole-genome sequence of the oriental lung fluke Paragonimus westermani. Gigascience 8, giy146.CrossRefGoogle ScholarPubMed
Ohari, Y, Suzuki, Y, Shibahara, T and Itagaki, T (2019) First report of Paragonimus skrjabini miyazakii metacercariae in Geothelphusa dehaani (Sawagani) occurring in Iwate Prefecture, Japan. Journal of Veterinary Internal Medicine 81, 11091112.Google ScholarPubMed
Perna, NT and Kocher, TD (1995) Patterns of nucleotide composition at fourfold degenerate sites of animal mitochondrial genomes. Journal of Molecular Evolution 41, 353358.CrossRefGoogle ScholarPubMed
Pham, KLT, Saijuntha, W, Lawton, SP and Le, TH (2022) Mitophylogenomics of the zoonotic fluke Echinostoma malayanum confirms it as a member of the genus Artyfechinostomum Lane, 1915 and illustrates the complexity of Echinostomatidae systematics. Parasitology Research 121, 899913.CrossRefGoogle ScholarPubMed
Qian, L, Zhou, P, Li, W, Wang, H, Miao, T and Hu, L (2018) Characterization of the complete mitochondrial genome of the lung fluke. Paragonimus heterotremus. Mitochondrial DNA Part B 3, 560561.CrossRefGoogle ScholarPubMed
Rajapakse, RPVJ, Pham, KLT, Karunathilake, KJK, Lawton, SP and Le, TH (2020) Characterization and phylogenetic properties of the complete mitochondrial genome of Fascioloides jacksoni (syn. Fasciola jacksoni) support the suggested intergeneric change from Fasciola to Fascioloides (Platyhelminthes: Trematoda: Plagiorchiida). Infection Genetics and Evolution 82, 104281.CrossRefGoogle Scholar
Rhoads, A and Au, KF (2015) PacBio sequencing and its applications. Genomics, Proteomics & Bioinformatics 13, 278289.CrossRefGoogle ScholarPubMed
Rosa, BA, Choi, YJ, McNulty, S, Jung, H, Martin, J, Agatsuma, T, Sugiyama, H, Le, TH, Doanh, PN, Maleewong, W, Blair, D, Brindley, PJ, Fischer, PU and Mitreva, M (2020) Comparative genomics and transcriptomics of four Paragonimus species provide insights into lung fluke parasitism and pathogenesis. Gigascience 9, giaa073.CrossRefGoogle Scholar
Shekhovtsov, SV, Katokhin, AV, Kolchanov, NA and Mordvinov, VA (2010) The complete mitochondrial genomes of the liver flukes Opisthorchis felineus and Clonorchis sinensis (Trematoda). Parasitology International 59, 100103.CrossRefGoogle Scholar
Singh, T, Hiromu, S, Devi, K and Singh, W (2015) First case of Paragonimus westermani infection in a female patient in India. Indian Journal of Medical Microbiology 33, 156159.CrossRefGoogle Scholar
Solà, E, Álvarez-Presas, M, Frías-López, C, Littlewood, DTJ, Rozas, J and Riutort, M (2015) Evolutionary analysis of mitogenomes from parasitic and free-living flatworms. PLOS One 10, e0120081.CrossRefGoogle ScholarPubMed
Talavera, G and Castresana, J (2007) Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Systematic Biology 56, 564577.CrossRefGoogle ScholarPubMed
Urabe, M and Marcaida, AJB (2022) A record of juvenile Paragonimus skrjabini miyazakii from the urinary bladder of the Japanese toad, Bufo japonicus formosus. Parasitology International 86, 102474.CrossRefGoogle ScholarPubMed
Voronova, AN, Vainutis, KS, Tabakaeva, TV, Sapotsky, MV, Karaeka, NN, Volkov, YG, Galkina, IV and Shchelkanov, MY (2022) Molecular identification of the trematode P. ichunensis stat. n. from lungs of Siberian tigers justified reappraisal of Paragonimus westermani species complex. Journal of Parasitic Diseases. https://doi.org/10.1007/s12639-022-01481-7.CrossRefGoogle ScholarPubMed
Wang, T, Wang, Y, Xu, F, Li, X, Qu, R, Song, L, Tang, Y and Lin, P (2018) Characterization of the complete mitochondrial genome of the lung fluke, Paragonimus kellicotti. Mitochondrial DNA Part B 3, 715716.CrossRefGoogle ScholarPubMed
Wey-Fabrizius, AR, Podsiadlowski, L, Herlyn, H and Hankeln, T (2013) Platyzoan mitochondrial genomes. Molecular Phylogenetics and Evolution 69, 365375.CrossRefGoogle ScholarPubMed
Fu, YT, Jin, YC, Li, F and Liu, GH (2019) Characterization of the complete mitochondrial genome of the echinostome Echinostoma miyagawai and phylogenetic implications. Parasitology Research 118(10), 30913097. https://doi.org/10.1007/s00436-019-06417-4.CrossRefGoogle ScholarPubMed
Lee, D, Choe, S, Park, H, Jeon, HK, Chai, JY, Sohn, WM, Yong, TS, Min, DY, Rim, HJ and Eom, KS (2013) Complete mitochondrial genome of Haplorchis taichui and comparative analysis with other trematodes. Korean Journal of Parasitology 51(6), 719726. https://doi.org/10.3347/kjp.2013.51.6.719.CrossRefGoogle ScholarPubMed
Littlewood, DT, Lockyer, AE, Webster, BL, Johnston, DA and Le, TH (2006) The complete mitochondrial genomes of Schistosoma haematobium and Schistosoma spindale and the evolutionary history of mitochondrial genome changes among parasitic flatworms. Molecular Phylogenetics and Evolution 39, 452467. https://doi.org/10.1016/j.ympev.2005.12.012.CrossRefGoogle ScholarPubMed
Ma, J, He, JJ, Zhou, CY, Sun, MM, Cevallos, W, Sugiyama, H, Zhu, XQ and Calvopiña, M (2019) Characterization of the mitochondrial genome sequences of the liver fluke Amphimerus sp. (Trematoda: Opisthorchiidae) from Ecuador and phylogenetic implications. Acta Tropica 195, 9096. https://doi.org/10.1016/j.actatropica.2019.04.025.CrossRefGoogle ScholarPubMed
Ma, J, Sun, MM, He, JJ, Liu, GH, Ai, L, Chen, MX and Zhu, XQ (2017) Fasciolopsis buski (Digenea: Fasciolidae) from China and India may represent distinct taxa based on mitochondrial and nuclear ribosomal DNA sequences. Parasites and Vectors 10(1), 101. https://doi.org/10.1186/s13071-017-2039-2.CrossRefGoogle ScholarPubMed
Na, L, Gao, JF, Liu, GH, Fu, X, Su, X, Yue, DM, Gao, Y, Zhang, Y and Wang, CR (2016) The complete mitochondrial genome of Metorchis orientalis (Trematoda: Opisthorchiidae): comparison with other closely related species and phylogenetic implications. Infection Genetics and Evolution 39, 4550. https://doi.org/10.1016/j.meegid.2016.01.010.CrossRefGoogle ScholarPubMed
Yang, X, Gasser, RB, Koehler, AV, Wang, L, Zhu, K, Chen, L, Feng, H, Hu, M and Fang, R (2015) Mitochondrial genome of Hypoderaeum conoideum – comparison with selected trematodes. Parasites and Vectors 8, 97. https://doi.org/10.1186/s13071-015-0720-x.CrossRefGoogle ScholarPubMed
Fu, YT, Jin, YC, Li, F and Liu, GH (2019) Characterization of the complete mitochondrial genome of the echinostome Echinostoma miyagawai and phylogenetic implications. Parasitology Research 118(10), 30913097. https://doi.org/10.1007/s00436-019-06417-4.CrossRefGoogle ScholarPubMed
Lee, D, Choe, S, Park, H, Jeon, HK, Chai, JY, Sohn, WM, Yong, TS, Min, DY, Rim, HJ and Eom, KS (2013) Complete mitochondrial genome of Haplorchis taichui and comparative analysis with other trematodes. Korean Journal of Parasitology 51(6), 719726. https://doi.org/10.3347/kjp.2013.51.6.719.CrossRefGoogle ScholarPubMed
Littlewood, DT, Lockyer, AE, Webster, BL, Johnston, DA and Le, TH (2006) The complete mitochondrial genomes of Schistosoma haematobium and Schistosoma spindale and the evolutionary history of mitochondrial genome changes among parasitic flatworms. Molecular Phylogenetics and Evolution 39, 452467. https://doi.org/10.1016/j.ympev.2005.12.012.CrossRefGoogle ScholarPubMed
Ma, J, He, JJ, Zhou, CY, Sun, MM, Cevallos, W, Sugiyama, H, Zhu, XQ and Calvopiña, M (2019) Characterization of the mitochondrial genome sequences of the liver fluke Amphimerus sp. (Trematoda: Opisthorchiidae) from Ecuador and phylogenetic implications. Acta Tropica 195, 9096. https://doi.org/10.1016/j.actatropica.2019.04.025.CrossRefGoogle ScholarPubMed
Ma, J, Sun, MM, He, JJ, Liu, GH, Ai, L, Chen, MX and Zhu, XQ (2017) Fasciolopsis buski (Digenea: Fasciolidae) from China and India may represent distinct taxa based on mitochondrial and nuclear ribosomal DNA sequences. Parasites and Vectors 10(1), 101. https://doi.org/10.1186/s13071-017-2039-2.CrossRefGoogle ScholarPubMed
Na, L, Gao, JF, Liu, GH, Fu, X, Su, X, Yue, DM, Gao, Y, Zhang, Y and Wang, CR (2016) The complete mitochondrial genome of Metorchis orientalis (Trematoda: Opisthorchiidae): comparison with other closely related species and phylogenetic implications. Infection Genetics and Evolution 39, 4550. https://doi.org/10.1016/j.meegid.2016.01.010.CrossRefGoogle ScholarPubMed
Yang, X, Gasser, RB, Koehler, AV, Wang, L, Zhu, K, Chen, L, Feng, H, Hu, M and Fang, R (2015) Mitochondrial genome of Hypoderaeum conoideum – comparison with selected trematodes. Parasites and Vectors 8, 97. https://doi.org/10.1186/s13071-015-0720-x.CrossRefGoogle ScholarPubMed
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