Main Article Content
Abstract
The present study is the first morphological and molecular characterization of a monogenean Eudiplozoon nipponicum (Goto, 1891) parasitizing gill filaments of common carps (Cyprinus carpio) obtained from the Lesser Zab River in northeastern Iraq in the subdistrict of Altun-Kopru from July to October 2022 and transported to the Laboratory of Zoology Research, Salahaddin University-Erbil, Iraq. Most previous studies regarding the considered parasite have targeted morphological data analysis. However, DNA sequence outline is typically supportive in systematics, since it contains aspects that are absent from morphological research. The main goal was to molecularly identify E. nipponicum utilizing the nuclear 28S rDNA region by PCR and nucleotide sequencing approach. The sequences were obtained and compared to the accessible GenBank sequences. The results justify the validation of E. nipponicum in Iraq using traditional (morphology-based) and modern (molecular-based) parasitological techniques. The latter one showed 99.11% (identity percentage) of E. nipponicum in comparison to the registered sequences of NCBI. Additionally, this was considered the first wide-ranging morphological and molecular study in the studied region. The phylogenetic relationship was concluded using Maximum Likelihood (ML) method. It was concluded that besides of the morphological characterization of E. nipponicum, PCR sequencing was considered a suitable molecular method for recognizing monogenean diplozoid species on the fishes.
Keywords
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
References
- Abdullah, S. M. A., & Nasraddin, M. O. (2015). Monogenean infections on some fishes from Lesser Zab River, Kurdistan Region, Iraq. American Journal of Biology and Life Sciences, 3(5), 161-167.
- http://www.openscienceonline.com/journal/archive2?journalId=704&paperId=2318
- Ahmadi, A., Borji, H., Naghibi, A., Nasiri, M. R., & Sharifiyazdi, H. (2017). Morphologic and molecular (28S rDNA characterization of Dactylogyrus spp. in Cyprinus carpio and Ctenopharyngodon idella in Mashhad, Iran. Canadian Journal of Veterinary Research, 81(4), 280-284.
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5644451/
- Al-Jubori, M. I. A., & Al-Nasiri, F. S. (2014). First record of two Paradiplozoon (Monogenea) from cyprinid fishes in Iraq. Jordan Journal of Agricultural Sciences, 10(4), 673-679.
- Al-Nasiri, F. S. (2003). First occurrence of the monogenetic trematode Diplozoon nipponicum Goto, 1891 in Iraq from common carp Cyprinus carpio (Pisces). Iraqi Journal of Agricultural Sciences, 8(6), 95-99.
- https://scholar.google.com/scholar?cluster=17619615970576148746&hl=en&oi=scholarr
- Al-Saadi, A. A. J., Mhaisen, F. T., & Hasan, H. R. (2010). Ectoparasites of seven fish species from Al-Husainia creek, Karbala province, mid Iraq. Journal of Kerbala University, 8(4), 1-7.
- https://www.iasj.net/iasj/article/69889
- Chmurciakova, N., Kasny, M., & Orosova, M. (2020). Cytogenetics of Eudiplozoon nipponicum (Monogenea, Diplozoidae): Karyotype, spermatocyte division and 18S rDNA location. Parasitology International, 76, 102031.
- https://doi.org/10.1016/j.parint.2019.102031
- Civáňová, K., Koyun, M., & Koubková, B. (2013). The molecular and morphometrical description of a new diplozoid species from the gills of the Garra rufa (Heckel, 1843) (Cyprinidae) from Turkey—including a commentary on taxonomic division of Diplozoidae. Parasitology Research, 112(8), 3053-3062.
- https://doi.org/10.1007/s00436-013-3480-6
- Coad, B. W. (2010). Freshwater Fishes of Iraq. Pensoft Publishers. Sofia-Moscow. 294 pp
- https://books.google.iq/books?id=xyXtRgAACAAJ
- Dodangeh, S., Fakhar, M., & Kialashaki, E. (2017). Application of molecular techniques for taxonomic and epidemiological studies of parasitic infections [Review]. Journal of Mazandaran University of Medical Sciences, 27(153), 163-174.
- http://jmums.mazums.ac.ir/article-1-9378-en.html
- Dos Santos, Q. M., & Avenant-Oldewage, A. (2020). Review on the molecular study of the Diplozoidae: analyses of currently available genetic data, what it tells us, and where to go from here. Parasites & Vectors, 13(1), 539.
- https://doi.org/10.1186/s13071-020-04417-3
- Dutra Vieira, T., Pegoraro de Macedo, M. R., Fedatto Bernardon, F., & Müller, G. (2017). Morphological, molecular and phylogenetic analyses of Diplotriaena bargusinica Skrjabin, 1917 (Nematoda: Diplotriaenidae). Parasitology International, 66(5), 555-559.
- https://doi.org/10.1016/j.parint.2017.04.009
- Dwivedi, S., Purohit, P., Misra, R., Pareek, P., Goel, A., Khattri, S., Pant, K. K., Misra, S., & Sharma, P. (2017). Diseases and molecular diagnostics: a step closer to precision medicine. Indian Journal of Clinical Biochemistry, 32(4), 374-398.
- https://doi.org/10.1007/s12291-017-0688-8
- Freyhof, J., Kaya, C., & Ali, A. (2021). A Critical Checklist of the Inland Fishes Native to the Euphrates and Tigris Drainages. In L. A. Jawad (Ed.), Tigris and Euphrates Rivers: Their Environment from Headwaters to Mouth (pp. 815-854). Springer International Publishing.
- https://doi.org/10.1007/978-3-030-57570-0_35
- Fricke, R., Eschmeyer, W. N., & Fong, J. D. (2023, 7 April, 2023). Eschmeyer's Catalog of Fishes. California Academy of Sciences. https://researcharchive.calacademy.org/research/ichthyology/catalog/SpeciesByFamily.asp
- Froese, R., & Pauly, D. (2022). FishBase. World Wide Web electronic publication. Retrieved (08/2022). from www.fishbase.org
- Gao, Q. X., M, C., Yao, W. J., Gao, Y., Song, Y., Wang, G. T., Wang, M. X., & Nie, P. (2007). Phylogeny of diplozoids in five genera of the subfamily Diplozoinae Palombi, 1949 as inferred from ITS-2 rDNA sequences. Parasitology, 134(Pt 5), 695-703.
- https://doi.org/10.1017/s0031182006001971
- Goto, S. (1891). On Diplozoon nipponicum n. sp. The Journal of the College of Science, Imperial University of Tokyo, 4, 151-192.
- Hebert, P. D. N., & Gregory, T. R. (2005). The Promise of DNA Barcoding for Taxonomy. Systematic Biology, 54(5), 852-859.
- https://doi.org/10.1080/10635150500354886
- Hodová, I., Sonnek, R., Gelnar, M., & Valigurová, A. (2018). Architecture of Paradiplozoon homoion: A diplozoid monogenean exhibiting highly-developed equipment for ectoparasitism. PLoS One, 13(2), e0192285.
- https://doi.org/10.1371/journal.pone.0192285
- Ilgova, J., Kavanova, L., Matiaskova, K., Salat, J., & Kasny, M. (2020). Effect of cysteine peptidase inhibitor of Eudiplozoon nipponicum (Monogenea) on cytokine expression of macrophages in vitro. Molecular and Biochemical Parasitology, 235, 111-248.
- https://doi.org/10.1016/j.molbiopara.2019.111248
- Khang, T. F., Soo, O. Y., Tan, W. B., & Lim, L. H. (2016). Monogenean anchor morphometry: systematic value, phylogenetic signal, and evolution. Peer Journal Life and Environment, 4, e1668.
- https://doi.org/10.7717/peerj.1668
- Khotenovsky, I. A. (1981). Taxonomy and phylogeny of monogenians of the families Diplozoidae and Discocotilidae (Monogenea). Parazitologiceskij Sbornik, 30, 166-178.
- https://eurekamag.com/research/021/856/021856493.php
- Koskova, E., Matejusova, I., Civanova, K., & Koubkova, B. (2010). Ethanol-fixed material used for both classical and molecular identification purposes: Eudiplozoon nipponicum (Monogenea: Diplozoidae) as a case parasite species. Parasitology Research, 107(4), 909-914.
- https://doi.org/10.1007/s00436-010-1949-0
- Koyee, Q. M. K., & Abdullah, S. M. A. (2019). Host Specificity, Community Components and Diversity Dynamics of Dactylogyrus spp. (Monogenean ectoparasites) Parasitizing Cyprinid Gills [journal article]. Polish Journal of Environmental Studies, 28(6), 4257-4269. https://doi.org/10.15244/pjoes/99064
- Koyee, Q. M. K., Khailany, R. A., Al-Marjan, K. S. N., & Abdullah, S. M. A. (2016). Molecular-Based Identification of Polystoma integerrimum by 28S rDNA, Phylogenetic and Secondary Structure Analysis Jordan Journal of Biological Sciences, 9(2), 117-121.
- Matejusová, I., Koubková, B., & Cunningham, C. O. (2004). Identification of European diplozoids (Monogenea, Diplozoinae) by restriction digestion of the ribosomal RNA internal transcribed spacer. Journal of Parasitology, 90(4), 817-822.
- https://doi.org/10.1645/ge-138r
- Matejusová, I., Koubková, B., Gelnar, M., & Cunningham, C. O. (2002). Paradiplozoon homoion Bychowsky & Nagibina, 1959 versus P. gracile Reichenbach-Klinke, 1961 (Monogenea): two species or phenotypic plasticity?. Systymatic Parasitology, 53(1), 39-47.
- https://doi.org/10.1023/a:1019945921143
- Mhaisen, F. T. (2023). Checklist of Iraqi Diplozoon species.
- Mhaisen, F. T., & Abdul-Ameer, K. N. (2014). Checklist of Diplozoid species (Monogenea) from Fishes of Iraq. Bulletin of the Iraq Natural History Museum, 13(2), 95-111.
- https://www.iasj.net/iasj/download/ca321de707173a44
- Mohammed, H. J. (2017). Parasitic fauna of some fish species from Diyala River in Diyala Province. [M.Sc. Thesis, University of Baghdad]. Baghdad. 22 pp ( In Arabic).
- Mohanta, U. K., & Itagaki, T. (2017). Molecular characterization and phylogeny of Linguatula serrata (Pentastomida: Linguatulidae) based on the nuclear 18S rDNA and mitochondrial cytochrome c oxidase I gene. The Journal of Veterinary Science, 79(2), 398-402.
- https://doi.org/10.1292/jvms.16-0508
- Mollaret, I., Jamieson, B. G., & Justine, J. L. (2000). Phylogeny of the monopisthocotylea and Polyopisthocotylea (Platyhelminthes) inferred from 28S rDNA sequences. International journal for parasitology, 30(2), 171-185.
- https://doi.org/10.1016/s0020-7519(99)00197-6
- Nishihira, T., & Urabe, M. (2020). Morphological and molecular studies of Eudiplozoon nipponicum (Goto, 1891) and Eudiplozoon kamegaii sp. n. (Monogenea; Diplozoidae). Folia Parasitologica, 67(1).
- https://doi.org/10.14411/fp.2020.018
- Olson, P. D., & Littlewood, D. T. J. (2002). Phylogenetics of the Monogenea--evidence from a medley of molecules. International Journal for Parasitology, 32(3), 233-244.
- https://doi.org/10.1016/s0020-7519(01)00328-9
- Pandey, K. C. (2010). An encyclopaedia of Indian Monogenoidea (1st ed.). Vitasta Publishing, New Delhi. 534 pp
- Patwardhan, A., Ray, S., & Roy, A. (2014). Molecular markers in phylogenetic studies- A review. Journal of Phylogenetics & Evolutionary Biology, 2(2), 1000131.
- https://doi.org/10.4172/2329-9002.1000131
- Rahemo, Z. I. F. (1980). Diplozoon kasimii new species from a freshwater teleost fish, Cyprinion macrostomum Heckel. Bulletin of the Biological Research Centre, 12(1), 109-114.
- Rana, N., & Das, B. K. (2016). Morphometric and molecular identification of Paradactylogyrus catlaius (Thapar 1948) in Catla catla (Hamilton 1822). Journal of Parasitic Diseases, 40(1), 36-40.
- https://doi.org/10.1007/s12639-014-0437-3
- Singh, H. S., & Chaudhary, A. (2010). Genetic characterization of Dactylogyroides longicirrus (Tripathi, 1959) Gussev, 1976 by nuclear 28S segment of ribosomal DNA with a morphological redescription. Scientia Parasitologica, 11(3), 119-127.
- Susurluk, A., Tarasco, E., Ehlers, R. U., & Triggiani, O. (2007). Molecular Characterization of Entomopathogenic Nematodes isolated in Italy by PCR-RFLP Analysis of the its region of Ribosomal DNA Repeat Unit. Nematologia Mediterranea, 35(1), 23-28.
- https://journals.flvc.org/nemamedi/article/view/86913
- Tamura, K., Stecher, G., & Kumar, S. (2021). MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology and Evolution, 38(7), 3022-3027.
- https://doi.org/10.1093/molbev/msab120
- Valigurová, A., Hodová, I., Sonnek, R., Koubková, B., & Gelnar, M. (2011). Eudiplozoon nipponicum in focus: monogenean exhibiting a highly specialized adaptation for ectoparasitic lifestyle. Parasitology Research, 108(2), 383-394.
- https://doi.org/10.1007/s00436-010-2077-6
References
Abdullah, S. M. A., & Nasraddin, M. O. (2015). Monogenean infections on some fishes from Lesser Zab River, Kurdistan Region, Iraq. American Journal of Biology and Life Sciences, 3(5), 161-167.
http://www.openscienceonline.com/journal/archive2?journalId=704&paperId=2318
Ahmadi, A., Borji, H., Naghibi, A., Nasiri, M. R., & Sharifiyazdi, H. (2017). Morphologic and molecular (28S rDNA characterization of Dactylogyrus spp. in Cyprinus carpio and Ctenopharyngodon idella in Mashhad, Iran. Canadian Journal of Veterinary Research, 81(4), 280-284.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5644451/
Al-Jubori, M. I. A., & Al-Nasiri, F. S. (2014). First record of two Paradiplozoon (Monogenea) from cyprinid fishes in Iraq. Jordan Journal of Agricultural Sciences, 10(4), 673-679.
Al-Nasiri, F. S. (2003). First occurrence of the monogenetic trematode Diplozoon nipponicum Goto, 1891 in Iraq from common carp Cyprinus carpio (Pisces). Iraqi Journal of Agricultural Sciences, 8(6), 95-99.
https://scholar.google.com/scholar?cluster=17619615970576148746&hl=en&oi=scholarr
Al-Saadi, A. A. J., Mhaisen, F. T., & Hasan, H. R. (2010). Ectoparasites of seven fish species from Al-Husainia creek, Karbala province, mid Iraq. Journal of Kerbala University, 8(4), 1-7.
https://www.iasj.net/iasj/article/69889
Chmurciakova, N., Kasny, M., & Orosova, M. (2020). Cytogenetics of Eudiplozoon nipponicum (Monogenea, Diplozoidae): Karyotype, spermatocyte division and 18S rDNA location. Parasitology International, 76, 102031.
https://doi.org/10.1016/j.parint.2019.102031
Civáňová, K., Koyun, M., & Koubková, B. (2013). The molecular and morphometrical description of a new diplozoid species from the gills of the Garra rufa (Heckel, 1843) (Cyprinidae) from Turkey—including a commentary on taxonomic division of Diplozoidae. Parasitology Research, 112(8), 3053-3062.
https://doi.org/10.1007/s00436-013-3480-6
Coad, B. W. (2010). Freshwater Fishes of Iraq. Pensoft Publishers. Sofia-Moscow. 294 pp
https://books.google.iq/books?id=xyXtRgAACAAJ
Dodangeh, S., Fakhar, M., & Kialashaki, E. (2017). Application of molecular techniques for taxonomic and epidemiological studies of parasitic infections [Review]. Journal of Mazandaran University of Medical Sciences, 27(153), 163-174.
http://jmums.mazums.ac.ir/article-1-9378-en.html
Dos Santos, Q. M., & Avenant-Oldewage, A. (2020). Review on the molecular study of the Diplozoidae: analyses of currently available genetic data, what it tells us, and where to go from here. Parasites & Vectors, 13(1), 539.
https://doi.org/10.1186/s13071-020-04417-3
Dutra Vieira, T., Pegoraro de Macedo, M. R., Fedatto Bernardon, F., & Müller, G. (2017). Morphological, molecular and phylogenetic analyses of Diplotriaena bargusinica Skrjabin, 1917 (Nematoda: Diplotriaenidae). Parasitology International, 66(5), 555-559.
https://doi.org/10.1016/j.parint.2017.04.009
Dwivedi, S., Purohit, P., Misra, R., Pareek, P., Goel, A., Khattri, S., Pant, K. K., Misra, S., & Sharma, P. (2017). Diseases and molecular diagnostics: a step closer to precision medicine. Indian Journal of Clinical Biochemistry, 32(4), 374-398.
https://doi.org/10.1007/s12291-017-0688-8
Freyhof, J., Kaya, C., & Ali, A. (2021). A Critical Checklist of the Inland Fishes Native to the Euphrates and Tigris Drainages. In L. A. Jawad (Ed.), Tigris and Euphrates Rivers: Their Environment from Headwaters to Mouth (pp. 815-854). Springer International Publishing.
https://doi.org/10.1007/978-3-030-57570-0_35
Fricke, R., Eschmeyer, W. N., & Fong, J. D. (2023, 7 April, 2023). Eschmeyer's Catalog of Fishes. California Academy of Sciences. https://researcharchive.calacademy.org/research/ichthyology/catalog/SpeciesByFamily.asp
Froese, R., & Pauly, D. (2022). FishBase. World Wide Web electronic publication. Retrieved (08/2022). from www.fishbase.org
Gao, Q. X., M, C., Yao, W. J., Gao, Y., Song, Y., Wang, G. T., Wang, M. X., & Nie, P. (2007). Phylogeny of diplozoids in five genera of the subfamily Diplozoinae Palombi, 1949 as inferred from ITS-2 rDNA sequences. Parasitology, 134(Pt 5), 695-703.
https://doi.org/10.1017/s0031182006001971
Goto, S. (1891). On Diplozoon nipponicum n. sp. The Journal of the College of Science, Imperial University of Tokyo, 4, 151-192.
Hebert, P. D. N., & Gregory, T. R. (2005). The Promise of DNA Barcoding for Taxonomy. Systematic Biology, 54(5), 852-859.
https://doi.org/10.1080/10635150500354886
Hodová, I., Sonnek, R., Gelnar, M., & Valigurová, A. (2018). Architecture of Paradiplozoon homoion: A diplozoid monogenean exhibiting highly-developed equipment for ectoparasitism. PLoS One, 13(2), e0192285.
https://doi.org/10.1371/journal.pone.0192285
Ilgova, J., Kavanova, L., Matiaskova, K., Salat, J., & Kasny, M. (2020). Effect of cysteine peptidase inhibitor of Eudiplozoon nipponicum (Monogenea) on cytokine expression of macrophages in vitro. Molecular and Biochemical Parasitology, 235, 111-248.
https://doi.org/10.1016/j.molbiopara.2019.111248
Khang, T. F., Soo, O. Y., Tan, W. B., & Lim, L. H. (2016). Monogenean anchor morphometry: systematic value, phylogenetic signal, and evolution. Peer Journal Life and Environment, 4, e1668.
https://doi.org/10.7717/peerj.1668
Khotenovsky, I. A. (1981). Taxonomy and phylogeny of monogenians of the families Diplozoidae and Discocotilidae (Monogenea). Parazitologiceskij Sbornik, 30, 166-178.
https://eurekamag.com/research/021/856/021856493.php
Koskova, E., Matejusova, I., Civanova, K., & Koubkova, B. (2010). Ethanol-fixed material used for both classical and molecular identification purposes: Eudiplozoon nipponicum (Monogenea: Diplozoidae) as a case parasite species. Parasitology Research, 107(4), 909-914.
https://doi.org/10.1007/s00436-010-1949-0
Koyee, Q. M. K., & Abdullah, S. M. A. (2019). Host Specificity, Community Components and Diversity Dynamics of Dactylogyrus spp. (Monogenean ectoparasites) Parasitizing Cyprinid Gills [journal article]. Polish Journal of Environmental Studies, 28(6), 4257-4269. https://doi.org/10.15244/pjoes/99064
Koyee, Q. M. K., Khailany, R. A., Al-Marjan, K. S. N., & Abdullah, S. M. A. (2016). Molecular-Based Identification of Polystoma integerrimum by 28S rDNA, Phylogenetic and Secondary Structure Analysis Jordan Journal of Biological Sciences, 9(2), 117-121.
Matejusová, I., Koubková, B., & Cunningham, C. O. (2004). Identification of European diplozoids (Monogenea, Diplozoinae) by restriction digestion of the ribosomal RNA internal transcribed spacer. Journal of Parasitology, 90(4), 817-822.
https://doi.org/10.1645/ge-138r
Matejusová, I., Koubková, B., Gelnar, M., & Cunningham, C. O. (2002). Paradiplozoon homoion Bychowsky & Nagibina, 1959 versus P. gracile Reichenbach-Klinke, 1961 (Monogenea): two species or phenotypic plasticity?. Systymatic Parasitology, 53(1), 39-47.
https://doi.org/10.1023/a:1019945921143
Mhaisen, F. T. (2023). Checklist of Iraqi Diplozoon species.
Mhaisen, F. T., & Abdul-Ameer, K. N. (2014). Checklist of Diplozoid species (Monogenea) from Fishes of Iraq. Bulletin of the Iraq Natural History Museum, 13(2), 95-111.
https://www.iasj.net/iasj/download/ca321de707173a44
Mohammed, H. J. (2017). Parasitic fauna of some fish species from Diyala River in Diyala Province. [M.Sc. Thesis, University of Baghdad]. Baghdad. 22 pp ( In Arabic).
Mohanta, U. K., & Itagaki, T. (2017). Molecular characterization and phylogeny of Linguatula serrata (Pentastomida: Linguatulidae) based on the nuclear 18S rDNA and mitochondrial cytochrome c oxidase I gene. The Journal of Veterinary Science, 79(2), 398-402.
https://doi.org/10.1292/jvms.16-0508
Mollaret, I., Jamieson, B. G., & Justine, J. L. (2000). Phylogeny of the monopisthocotylea and Polyopisthocotylea (Platyhelminthes) inferred from 28S rDNA sequences. International journal for parasitology, 30(2), 171-185.
https://doi.org/10.1016/s0020-7519(99)00197-6
Nishihira, T., & Urabe, M. (2020). Morphological and molecular studies of Eudiplozoon nipponicum (Goto, 1891) and Eudiplozoon kamegaii sp. n. (Monogenea; Diplozoidae). Folia Parasitologica, 67(1).
https://doi.org/10.14411/fp.2020.018
Olson, P. D., & Littlewood, D. T. J. (2002). Phylogenetics of the Monogenea--evidence from a medley of molecules. International Journal for Parasitology, 32(3), 233-244.
https://doi.org/10.1016/s0020-7519(01)00328-9
Pandey, K. C. (2010). An encyclopaedia of Indian Monogenoidea (1st ed.). Vitasta Publishing, New Delhi. 534 pp
Patwardhan, A., Ray, S., & Roy, A. (2014). Molecular markers in phylogenetic studies- A review. Journal of Phylogenetics & Evolutionary Biology, 2(2), 1000131.
https://doi.org/10.4172/2329-9002.1000131
Rahemo, Z. I. F. (1980). Diplozoon kasimii new species from a freshwater teleost fish, Cyprinion macrostomum Heckel. Bulletin of the Biological Research Centre, 12(1), 109-114.
Rana, N., & Das, B. K. (2016). Morphometric and molecular identification of Paradactylogyrus catlaius (Thapar 1948) in Catla catla (Hamilton 1822). Journal of Parasitic Diseases, 40(1), 36-40.
https://doi.org/10.1007/s12639-014-0437-3
Singh, H. S., & Chaudhary, A. (2010). Genetic characterization of Dactylogyroides longicirrus (Tripathi, 1959) Gussev, 1976 by nuclear 28S segment of ribosomal DNA with a morphological redescription. Scientia Parasitologica, 11(3), 119-127.
Susurluk, A., Tarasco, E., Ehlers, R. U., & Triggiani, O. (2007). Molecular Characterization of Entomopathogenic Nematodes isolated in Italy by PCR-RFLP Analysis of the its region of Ribosomal DNA Repeat Unit. Nematologia Mediterranea, 35(1), 23-28.
https://journals.flvc.org/nemamedi/article/view/86913
Tamura, K., Stecher, G., & Kumar, S. (2021). MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology and Evolution, 38(7), 3022-3027.
https://doi.org/10.1093/molbev/msab120
Valigurová, A., Hodová, I., Sonnek, R., Koubková, B., & Gelnar, M. (2011). Eudiplozoon nipponicum in focus: monogenean exhibiting a highly specialized adaptation for ectoparasitic lifestyle. Parasitology Research, 108(2), 383-394.