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In the Sea of Japan and the Adriatic Sea, a COX1 DNA test revealed genetic similarity of Manila clams, although shell parameters, spermatogenesis patterns, and sperm characteristics are area specific

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Abstract

The two Manila clam forms living in the Sea of Japan and in the Adriatic Sea were compared genetically and morphologically to identify traits that could be used to authenticate these geographic forms. COX1 DNA barcoding confirmed that both forms of these venerid bivalve belong to the species Ruditapes philippinarum (Adams and Reeve, 1850). It has also been shown that, based on COX1 analysis, it is not possible to separate these forms into different clusters based on geographic origin. However, some morphological features make it possible to differentiate these forms. It is noted that the contour of the shells of the Adriatic and Pacific molluscs is rounded and oblong, respectively. Underwater analysis of Pacific clams showed that shell color may be specific to specimens found at certain collection sites and differ between specimens taken from different collection sites. In the spermatogenesis of Adriatic and Pacific clams, a difference was found in the cellular mechanism of interaction between germplasm granules and mitochondria, manifested by distant contact and direct contact. Spermiogenesis has three similar lines of acrosome development, but differs in the number of ways in which the nucleus is formed. Due to the variability of the ways of sperm formation, heteromorphic morphs of spermatozoa are formed, the total number of which is six for the species R. philippinarum. In the Adriatic Manila clam, five morphs from this set are expressed. The Pacific Manila clam has only three sperm morphs. Only two sperm morphs are similar in both forms. In addition, each form is distinguished by the presence of unique sperm morphs and each form has its own type of dominant spermatozoon. Thus, the contour and color of the shells, as well as cytologic markers that have been found in the mechanisms of differentiation of meiotic cells and the characteristics of spermatozoa, can be used to distinguish the Adriatic and Pacific forms of the bivalve mollusc R. philippinarum.

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Data availability

The datasets generated and analysed during the current study are available from the corresponding author on request, and will be available in ResearchGate after publication of the article.

References

  • Au D, Reunov A, Wu S (1998) Four lines of spermatid development and dimorphic spermatozoa in the sea urchin Anthocidaris crassispina (Echinodermata, Echinoida). Zoomorphology 118:159–168. https://doi.org/10.1007/s004350050065

    Article  Google Scholar 

  • Bianchini G, Brombin V, Carlino P, Mistri E, Natali C, Salani GM (2021) Traceability and authentication of Manila clams from North-Western Adriatic Lagoons Using C and N Stable Isotope Analysis. Molecules 26:1859. https://doi.org/10.3390/molecules26071859

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Borowsky R, Luk A, He X, Kim RS (2018) Unique sperm haplotypes are associated with phenotypically different sperm subpopulations in Astyanax fish. BMC Biol 16:72. https://doi.org/10.1186/s12915-018-0538-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bourne N (1982) Distribution, reproduction and growth of Manila clam, Tapes philippinarum (Adams and Reeve, 1850), in British Columbia. J Shellfish Res 2:47–54

    Google Scholar 

  • Caill-Milly N, Bru N, Barranger M, Gallon L, D’amico F (2014) Morphological trends of four Manila clam populations (Venerupis philippinarum) on the French Atlantic coast: identified spatial patterns and their relationship to environmental variability. J Shellfish Res 33(2):355–372

    Article  Google Scholar 

  • Chetoui I, Denis F, Boussaid M, Telahigue K, El Cafsi M (2016) Genetic diversity and phylogenetic analysis of two Tunisian bivalves (Mactridae) Mactra corallina (Linnaeus, 1758) and Eastonia rugosa (Helbling, 1799) based on COI gene sequences. C R Biol 339:115–122. https://doi.org/10.1016/j.crvi.2016.02.001

    Article  PubMed  Google Scholar 

  • Chiesa S, Lucentini L, Freitas R, Nonnis Marzano F, Breda S, Figueira E, Caill-Milly N, Herbert RJH, Soares AMVM, Argese E (2016) Genetic diversity of introduced Manila clam. Bull Jap Fish Res Edu Agen 42:55–65

    Google Scholar 

  • Cordero D, Delgado M, Liu B, Ruesink J, Saavedra C (2017) Population genetics of the Manila clam (Ruditapes philippinarum) introduced in North America and Europe. Sci Rep 7:39745. https://doi.org/10.1038/srep39745

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ding J, Wen Q, Huo Z, Hongtao H, Qin Y, Yan X (2021) Identification of shell-color-related microRNAs in the Manila clam Ruditapes philippinarum using high-throughput sequencing of small RNA transcriptomes. Sci Rep 11:8044. https://doi.org/10.1038/s41598-021-86727-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fava G, Meggiato L (1995) Genetica biochimica e caratteri morfologici in popolazioni alto adriatiche di Ruditapes philippinarum e R. decussatus. Technical Report Ricerche e Sperimentazioni, Programma Integrato Mediterraneo per le zone lagunari dell'Adriatico settentrionale, pp 203–209

  • Gwo J-C, Huang Y-S, Kuo T-Y (2021) Sperm ultrastructure of Ruditapes variegata and Tapes literatus (Mollusca, Bivalvia, Veneridae, Tapetinae) from Pescadores, Taiwan. Tissue Cell 71:101575. https://doi.org/10.1016/j.tice.2021.101575

  • Katoh K, Standley DM (2013) MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability. Mol Biol Evol 30(4):772–780. https://doi.org/10.1093/molbev/mst010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim JH, Chung JS, Lee K-Y (2013) Ultrastructural characteristics of the testis, spermatogenesis and taxonomic values of sperm morphology in male Ruditapes philippinarum in Western Korea. Dev Reprod 17(2):121–132. https://doi.org/10.12717/DR.2013.17.2.121

    Article  PubMed  PubMed Central  Google Scholar 

  • Kim EM, Song MS, Hur DH, An CM, Kang JH, Park JY (2015) Easy method for discriminating the origins of Manila clam Ruditapes philippinarum with a dual-labelled PNA-probe-based melting curve analysis. Biochip J 9:247–258. https://doi.org/10.1007/s13206-015-9402-1

    Article  CAS  Google Scholar 

  • Leigh JW, Bryant D (2015) PopART: Full-feature software for haplotype network construction. Methods Ecol Evol 6(9):1110–1116. https://doi.org/10.1111/2041-210X.12410

    Article  Google Scholar 

  • Levitan D (2000) Sperm velocity and longevity trade off each other and influence fertilization in the sea urchin Lytechinus variegates. Proc R Soc Lond B 267:531–534

    Article  CAS  Google Scholar 

  • Low M, Pkl Ng, Clark PF (2020) Additional notes on the publication of the narrative, zoology and notes from a journal of research into the natural history of the voyage of HMS Samarang and its consequences for the nomenclature of decapod crustaceans and other taxa. Zootaxa 4809(2):zootaxa.4809.2.3. https://doi.org/10.11646/zootaxa.4809.2.3

    Article  PubMed  Google Scholar 

  • Markaide P, Gairín I, Cordero D, Ibarrola I, Saavedra C (2021) No hybridization and marked interspecific differences in individual growth rate in mixed cultures of Manila clam (Ruditapes philippinarum) and grooved carpet-shell clam (R. decussatus). Aquaculture 541:736824. https://doi.org/10.1016/j.aquaculture.2021.736824

    Article  CAS  Google Scholar 

  • Minh BQ, Schmidt HA, Chernomor O, Schrempf D, Woodhams MD, von Haeseler A, Lanfear R (2020) IQ-TREE 2: New models and efficient methods for phylogenetic inference in the genomic era. Mol Biol Evol 37:1530–1534. https://doi.org/10.1093/molbev/msaa015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Morgan C (2002) Naturalisation of an introduced species, the Manila clam, Tapes philippinarum, in Poole Harbour, UK. Spec Publ – Aquac Assoc Can 6:53–55

    Google Scholar 

  • Morrow EH, Gage MJG (2001) Consistent significant variation between individual males in spermatozoal morphometry. J Zool 254(2):147–153

    Article  Google Scholar 

  • Mortensen SH (1993) A health survey of selected stocks of commercially exploited Norwegian bivalve molluscs. Dis Aquat Org 16(2):149–156

    Article  Google Scholar 

  • Morton B (1976) The biology, ecology and functional aspects of the organs of feeding and digestion of the S.E. Asian mangrove bivalve, Enigmonia aenigmatica (Mollusca: Anomiacea). J Zool Lond 179:437–466

    Article  Google Scholar 

  • Moss SM (1990) Enigmonia aenigmatica: an enigmatic molluscan chameleon. The marine biology of the South China Sea. In: Morton B (ed) Proceedings of the first international conference of the marine biology of Hong Kong and the South China Sea. Hong Kong, 28 October – 3 November 1990, Hong Kong University Press, Hong Kong

  • Ni G, Li Q, Lehai Ni, Kong L, Yu H (2015) Population subdivision of the surf clam Mactra chinensis in the East China Sea: Changjiang River outflow is not the sole driver. PeerJ 3:e1240. https://doi.org/10.7717/peerj.1240

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nosho TY, Chew KK (1972) The setting and growth of the Manila clam, Venerupis japonica (Deshayes), in Hood Canal, Washington. Proc Natl Shellfish Assoc 62:50–58

    Google Scholar 

  • Ponurovsky SK, Selin NI (1988) Distribution, population structure, and growth of the bivalve molusk Ruditapes philippinarum in Vostok Bay, Sea of Japan. Sov J Mar Biol 1:11–15

    Google Scholar 

  • Reunov A, Au D, Wu R (1999) Spermatogenesis of the green-lipped mussel Perna viridis with dual patterns of acrosome and tail development in spermatids. Helgol Mar Res 53:62–69

    Article  Google Scholar 

  • Reunov A, Vekhova E, Zakharov E, Reunova Y, Alexandrova Y, Sharina S, Adrianov A (2018) Variation of sperm morphology in Pacific oyster precludes its use as a species marker but enables intraspecific geo-authentification and aquatic monitoring. Helgol Mar Res 72:8. https://doi.org/10.1186/s10152-018-0510-x

    Article  Google Scholar 

  • Reunov A, Alexandrova Y, Reunova Y, Komkova A, Milani L (2019a) Germ plasm provides clues on meiosis: the concerted action of germ plasm granules and mitochondria in gametogenesis of the clam Ruditapes philippinarum. Zygote 27(1):25–35. https://doi.org/10.1017/S0967199418000588

    Article  CAS  PubMed  Google Scholar 

  • Reunov A, Yakovlev K, Hu J, Reunova Y, Komkova A, Alexandrova Y, Pimenova E, Tiefenbach J, Krause H (2019b) Close association between vasa-positive germ plasm granules and mitochondria correlates with cytoplasmic localization of 12S and 16S mtrRNAs during zebrafish spermatogenesis. Differentiation 109:34–41. https://doi.org/10.1016/j.diff.2019.08.002

    Article  CAS  PubMed  Google Scholar 

  • Reunov A, Alexandrova Y, Komkova A, Reunova Y, Pimenova E, Vekhova E, Milani L (2021a) VASA-induced cytoplasmic localization of CYTB-positive mitochondrial substance occurs by destructive and nondestructive mitochondrial effusion, respectively, in early and late spermatogenic cells of the Manila clam. Protoplasma 258(4):817–825. https://doi.org/10.1007/s00709-020-01601-1

    Article  CAS  PubMed  Google Scholar 

  • Reunov A, Lutaenko K, Vekhova E, Zhang J, Zakharov E, Sharina S, Alexandrova Y, Reunova Y, Akhmadieva A, Adrianov A (2021b) In the Asia-Pacific region, the COI DNA test revealed the divergence of the bivalve mollusc Mactra chinensis into three species; can these species be distinguished using shell coloration and sperm structure? Helgol Mar Res 75:7. https://doi.org/10.1186/s10152-021-00553-0

    Article  Google Scholar 

  • Sekine Y, Yamakawa Y, Takazawa S, Yingping L, Toba M (2006) Geographic variation in the COXI gene of the Short-neck clam Ruditapes philippinarum in coastal regions of Japan and China. Venus 65:229–224

    Google Scholar 

  • Sigurdson JB, Sundari G (1990) Shellular changes in the colour in the tree-climbing bivalve Enigmonia aenigmatica (Holten 1802) (Annomiidae). Raffles Bull Zool 38:213–218

    Google Scholar 

  • Silina AV (2010) Population characteristics of the bivalve Ruditapes philippinarum from Cheju Island coasts, Korea. Korean J Malacol 26(3):227–234

    Google Scholar 

  • Skurikhina LA, Kartavtsev YF, Chichvarkhin AY, Pankova MV (2001) Study of two species of mussels, Mytilus trossulus and Mytilus galloprovinciallis (Bivalvia: Mytilidae), and their hybrids in Peter the Great Bay of the Sea of Japan with the use of PCR markers. Genetika 37:1717–1720

    CAS  PubMed  Google Scholar 

  • Sokolova IM, Berger VJ (2000) Physiological variation related to shell colour polymorphism in White Sea Littorina saxatilis. J Exp Mar Biol Ecol 245(1):1–23. https://doi.org/10.1016/S0022-0981(99)00132-X

    Article  Google Scholar 

  • Tan Y, Fang L, Qiu M, Huo Z, Yan X (2020) Population genetics of the Manila clam (Ruditapes philippinarum) in East Asia. Sci Rep 10:21890. https://doi.org/10.1038/s41598-020-78923-w

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tomiyama T (2021) Ecophenotypic plasticity in shell growth direction of asari clam Ruditapes philippinarum. J Mar Biol Assoc United Kingdom 101(3):555–560. https://doi.org/10.1017/S0025315421000412

    Article  CAS  Google Scholar 

  • Williams ST (2017) Molluscan shell colour. Biol Rev 92:1039–1058. https://doi.org/10.1111/brv.12268

    Article  PubMed  Google Scholar 

  • Yan X, Nie H, Huo Z et al (2019) Clam genome sequence clarifies the molecular basis of its benthic adaptation and extraordinary shell color diversity. iScience 19:1225–1237. https://doi.org/10.1016/j.isci.2019.08.049

    Article  PubMed  PubMed Central  Google Scholar 

  • Yuan H, Xu X, Yang F, Zhao L, Yan X (2020) Impact of seawater acidification on shell property of the Manila clam Ruditapes philippinarum grown within and without sediment. J Oceanol Limnol 38(1):236–248. https://doi.org/10.1007/s00343-019-8281-z

    Article  Google Scholar 

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Acknowledgements

Many thanks to Diving Department Head A. Goloseev and to divers N. Prohorov, K. Dudka, A. Oskolkov, M. Eremenko, I. Krasilnikov, S. Izrailskiy (National Scientific Centre of Marine Biology, Vladivostok, Russia) for very professional collection of R. philippinarum. Many thanks to Elizabeth Alexandrov for photographing mollusc shells. We are deeply indebted to Denis Fomin for the helpful assistance during observations with the scanning electron microscope and transmission electron microscope in the Microscopy Core Facility of the National Scientific Centre of Marine Biology (Vladivostok, Russia). A part of the electron microscopy study was done in the Microscopy Facility at St. Francis Xavier University (Antigonish, NS, Canada).

Funding

Part of the genetic testing was funded by the Italian Ministry of Education, University and Research MIUR-SIR Programme (Grant Number RBSI14G0P5) funded to LM.

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Author notes

  1. Arkadiy Reunov

    Present address: Department of Biology, St. Francis Xavier University, Antigonish, NS, B2G 2W5, Canada

Authors and Affiliations

  1. A.V. Zhirmunsky National Scientific Center of Marine Biology (NSCMB), Russian Academy of Sciences, Vladivostok, 690041, Russia

    Arkadiy Reunov, Evegenia Vekhova, Yana Alexandrova, Yulia Reunova, Anna Akhmadieva & Eugenia Pimenova

  2. Department of Biological, Geological and Environmental Sciences (BiGeA), University of Bologna (Italy), Bologna, Italy

    Liliana Milani, Giovanni Piccinini & Mariangela Iannello

  3. Canadian Centre for DNA Barcoding, Biodiversity Institute of Ontario, Guelph, Canada

    Evgeny Zakharov

  4. Far Eastern Federal University, School of Natural Sciences, 10 Ajax Bay, Russky Island, Vladivostok, 690950, Russia

    Anna Akhmadieva

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Contributions

All authors contributed to the study. AR conceived, design the study, analyzed the data, wrote the paper. LM analyzed the data, revised the paper. GP, MI collected the samples, performed genetic analyses, revised the paper. YR participated in the work using scanning electron microscopy and performed statistical analyses. EZ participated in genetic analyses. EV, YA, AA, EP collected samples, performed routine work using scanning electron microscopy and transmission electron microscopy. All authors read and approved the final version of the manuscript.

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Correspondence to Arkadiy Reunov.

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Reunov, A., Vekhova, E., Milani, L. et al. In the Sea of Japan and the Adriatic Sea, a COX1 DNA test revealed genetic similarity of Manila clams, although shell parameters, spermatogenesis patterns, and sperm characteristics are area specific. Biologia 78, 2803–2817 (2023). https://doi.org/10.1007/s11756-023-01414-6

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