Translator Disclaimer
1 January 2020 A Tropical Biodiversity Hotspot Under the New Threat: Discovery and DNA Barcoding of the Invasive Chinese Pond Mussel Sinanodonta Woodiana in Myanmar
Author Affiliations +
Abstract

A well-established invasive Sinanodonta woodiana population was discovered in a floodplain lake of the upstream section of the Irrawaddy River basin, Kachin State, Myanmar. The DNA barcoding reveals that the population belongs to the temperate invasive mtDNA lineage and represents the same cytochrome c oxidase subunit I haplotype, which has been recorded in the invasive European populations. It is the most southern location of a population appertaining to this highly invasive haplotype known to date. The actual distribution of this alien species in Myanmar is still unknown, although it appears to be rather not widespread. Its possible dispersal through the country may affect native benthos communities, which include many unique endemic taxa. However, further expansion of the temperate lineage across South East Asia is supposed to be limited due to a specific environment of tropical floodplain freshwater systems that appears too warm for this cryptic taxon.

Introduction

The Chinese pond mussel, Sinanodonta woodiana (Lea, 1834) is a well-known invasive species of the Unionidae, which is widely spread almost around the world together with its alien fish hosts (Douda, Vrtílek, Slavík, & Reichard, 2012; Lajtner & Crnčan, 2011; Lopes-Lima et al., 2017; Watters, 1997). Recent molecular studies reveal that S. woodiana is rather a complex of several closely related species because it comprises at least seven deeply divergent mtDNA lineages (Bolotov et al., 2016). Among them, the two lineages could only be considered as invasive. The first one is the tropical invasive lineage, which is currently widespread across the Malay Peninsula, the Philippines, and through Indonesia to the Lesser Sunda Islands (Bolotov et al., 2016; Zieritz et al., 2016). The origin of this successful invader is uncertain, but several authors placed it within Taiwan or the southern regions of continental China based on the analysis of primary sources for introduced fishes (Bolotov et al., 2016; Djajasasmita, 1982; Watters, 1997). The second one is the temperate invasive lineage, which has broad nonnative range in Europe, but most likely originated from the Yangtze drainage basin (Bolotov et al., 2016; Watters, 1997).

S. woodiana may affect native mussel populations, including the negative impact via cross-resistance of host fishes (Donrovich et al., 2017; Sousa, Novais, Costa, & Strayer, 2014). Adult Sinanodonta mussels can compete effectively with indigenous taxa for space, food, and host fishes. Their ability to modify natural ecosystems influences biological, physical, and chemical parameters of water environment (Bolotov et al., 2016; Douda et al., 2012; Guarneri et al., 2014; Sousa, Gutiérrez & Aldridge, 2009, 2014; Watters, 1997). An infection of fishes with the glochidia of Sinanodonta may negatively affect their physiological conditions, size, and weight (Douda et al., 2017).

In the present correspondence, we report the first occurrence of S. woodiana from Myanmar. In addition, we discuss the possibility of the further spread of this lineage of S. woodiana species complex across river systems of Southeast Asia and its possible effects on the native freshwater communities.

Methods

Data Collection

Available cytochrome c oxidase subunit I (COI) sequences of S. woodiana species complex and its sister taxa were downloaded from the BOLD IDS and NCBI’s GenBank, resulting in 68 sequences from Europe, China, Russia, South Korea, Japan, Malaysia, the Philippines, and Indonesia (Table 1). The COI sequence, which was provided for the S. aff. woodiana specimen from Romania (NCBI’s GenBank acc. no. JQ435822), was not included (see Bolotov et al., 2016). We have analyzed three available specimens of S. woodiana, which were collected from a floodplain lake of the upstream section of the Irrawaddy River basin, Kachin State, Myanmar in November 2016 during the join fieldworks with specialists of Flora & Fauna International and Department of Fisheries of Myanmar (Table 1). In addition, six new samples of S. woodiana complex from Vietnam and Russia were also sequenced. Sequences of Margaritifera dahurica and M. laosensis were used as out-groups (GenBank acc. nos. KJ161530 and KR006699, respectively).

Table 1.

List of the Mitochondrial COI Sequences ofSinanodonta spp. Examined in the Present Study.

10.1177_1940082917738151-table1.tif

Laboratory Protocols and Phylogenetic Analysis

A total genomic DNA was extracted from the alcohol-preserved mussel foot tissue using the NucleoSpin® Tissue Kit (Machereye Nagel GmbH & Co. KG, Germany), following the manufacturer’s protocol. Primers used for amplification of the COI partial sequences were LCO1490 and HCO 2198 (Folmer, Black, Hoeh, Lutz, & Vrijenhoek, 1994). The polymerase chain reaction (PCR) mix contained approximately 200 ng of total cellular DNA, 10 pmol of each primer, 200 mmol of each dNTP, 2.5 ml of PCR buffer (with 10 × 2 mmol MgCl2), 0.8 units of Taq DNA polymerase (SibEnzyme Ltd., Russia), and H2O, which was added up to a final volume of 25 ml. Thermocycling included one cycle at 95℃ (4 min), followed by 34 cycles at 95℃ (45 s), 50℃ (40 s), and 72℃ (50 s) with a final extension at 72℃ (5 min). Forward and reverse sequence reactions were performed directly on purified PCR products using the ABIPRISM® BigDye™ Terminator v. 3.1 reagents kit and run on an ABI PRISM® 3730 DNA analyzer (Thermo Fisher Scientific Inc., Waltham, MA, USA). The resulting sequences were checked using a sequence alignment editor (BioEdit v. 7.2.5; Hall, 1999).

The alignment of the COI sequences was performed using the ClustalW algorithm (Thompson, Higgins, & Gibson, 1994). For the phylogenetic analyses, each COI sequence of aligned data set was trimmed, leaving a 659-bp fragment. Then, identical COI sequences were removed from the data set using an online FASTA sequence toolbox (FaBox1.41; Villesen, 2007), leaving 35 haplotype sequences (including the two out-group taxa). For phylogenetic analyses, we used the COI data set with unique haplotypes. The best evolutional models for each partition calculated on the base of corrected Akaike Information Criterion through MEGA6 (Tamura, Stecher, Peterson, Filipski, & Kumar, 2013) were as follows: (a) first codon of the COI: HKY; (b) second codon of the COI: TN93 + G (G = 0.21); (c) third codon of the COI: HKY. Phylogenetic relationships were reconstructed based on Bayesian inference using MrBayes v. 3.2.6 (Ronquist et al., 2012) at the San Diego Supercomputer Center through the CIPRES Science Gateway (Miller, Pfeiffer, & Schwartz, 2010). Four Markov chains, one cold and three heated (temperature = 0.1), were run simultaneously for 25,000,000 generations. The resulting phylogeny was constructed using a tree figure drawing tool (Archaeopteryx v. 0.9901 beta; Han & Zmasek, 2009).

Results

A well-established population of S. aff. woodiana was discovered in a floodplain lake of the upstream section of the Irrawaddy River basin, Kachin State, Myanmar (Figures 1Figure 2. to 3). The mean shell length is 110.3 mm (90.2–128.5 mm, N = 20).

Figure 1.

Map of our new record (red asterisk) of the temperate invasive lineage of Sinanodonta aff. woodiana from Myanmar.

10.1177_1940082917738151-fig1.tif

Figure 2.

Habitat of Sinanodonta aff. woodiana in Myanmar: A floodplain lake near Bhamo, the Irrawaddy River drainage basin, Kachin State, Myanmar.

10.1177_1940082917738151-fig2.tif

Figure 3.

Shells of Sinanodonta aff. woodiana from a floodplain lake near Bhamo, the Irrawaddy River drainage basin, Kachin State, Myanmar: (a) Specimen no. RMBH biv269/15*, (b) Specimen no. RMBH biv269/16*, (c) Specimen no. RMBH biv269/9, and (d) Specimen no. RMBH biv269/6. An asterisk indicates sequenced specimens (Table 1). Scale bar = 2 cm.

10.1177_1940082917738151-fig3.tif

The three sequenced specimens of S. aff. woodiana belong to a single COI haplotype, which is identical to that recorded from invasive populations in Europe and from native populations in China (Figure 4). This haplotype belongs to the temperate invasive lineage of Bolotov et al. (2016), which also comprises three additional haplotypes from China. The mean COI p-distance within the temperate invasive lineage varies from 0.15% to 0.66%.

Figure 4.

The majority-rule consensus phylogenetic tree of Sinanodonta spp. and sister taxa recovered from Bayesian inference analysis of an alignment comprising 33 unique COI haplotypes. In addition, haplotypes of Margaritifera dahurica and M. laosensis were used as out-groups (GenBank acc. nos. KJ161530 and KR006699, respectively). Numbers near nodes are Bayesian posterior probabilities. Haplotype codes and lineage codes correspond to Table 1. The scale bar indicates the branch length.

10.1177_1940082917738151-fig4.tif

Populations from Malaysia, Indonesia, and Philippines belong to the tropical invasive lineage of Bolotov et al. (2016; see Figure 4). The sequenced specimens from Singapore are also representatives of this lineage, although they were obtained from ornamental trade (Ng et al., 2016). In contrast, the populations from Vietnam belong to a separate lineage that appears to be a native species, which inhabits the Red River basin (see Figure 4).

Discussion

Among countries of Southeast Asia, representatives of S. woodiana species complex were recorded from Malaysia, Indonesia, Singapore, Philippines, Cambodia, and Vietnam but were unknown from Myanmar, Thailand, and Laos (Bolotov et al., 2016; Ng et al., 2016; Zieritz et al., 2016, 2017).

Our sequenced specimens of S. woodiana from Irrawaddy basin surprisingly reveal the same COI haplotype that was recorded from invasive European populations and that belongs to the temperate lineage of Bolotov et al. (2016).

The taxonomy of S. woodiana species complex is unclear because it includes several divergent mtDNA lineages, each of which may represent a separate cryptic species (Bolotov et al., 2016; Froufe et al., 2017). Moreover, there is a plethora of nominal taxa that were synonymized with S. woodiana. With respect to an integrative approach (Konopleva, Bolotov, Vikhrev, Gofarov, & Kondakov, 2017), a taxonomic revision of such a species complex should be based on molecular sequence data obtained from the topotypes of each nominal taxon because old museum lots with dry shells are not appropriate for the extraction of high-quality DNA. The holotype of S. woodiana was collected from the Pearl River near Guangzhou (USNM: voucher no. 86380, the type locality: “Canton, China”), but any Sinanodonta sequences from this region, which may serve as reference sequences for the species, are not available.

Finally, the genus Sinanodonta should be a focus of the extensive taxonomic revision, along with efforts to estimate the distribution of invasive lineages and to detect the environmental threats and risks from alien mussel impacts.

Implications for Conservation

The actual distribution of S. woodiana in Myanmar is still unknown, although it seems to be rather local, because it was not recorded anywhere during our extensive fieldworks across the country (Bolotov et al., 2017a, 2017b). However, its possible spread through the country may affect native benthos communities, which include many unique endemic taxa of the Unionidae (Bolotov et al., 2017b). Our survey in the Bhamo area has shown that small lakes, which harbor the majority of mussel species in this region, may represent a suitable habitat for S. woodiana. The Chinese pond mussel could successfully inhabit these waterbodies and could compete with native mussels for resources and host fishes (Bolotov et al., 2016; Zieritz et al., 2016). Negative effects of the invasive S. woodiana on freshwater ecosystem functioning have been reported from Europe and Malaysia (Donrovich et al., 2017; Sousa et al., 2014; Zieritz et al., 2016). In Malaysia, this alien species replaced the native unionid assemblages in many river systems (Zieritz et al., 2016). However, we assume that further expansion of the temperate lineage across Southeast Asia may be limited due to the specific environment of tropical floodplain freshwater systems. These environmental conditions appear too warm for this cryptic taxon, which most likely originated from temperate China (see Figure 4). In contrast, a possible invasion of the tropical lineage into large freshwater basins of Myanmar (e.g., the Irrawaddy, Salween, and Sittaung drainage basins) seems to be much more dangerous because this lineage is adapted well to the environment of tropical monsoon freshwater systems, and it may therefore spread throughout the country.

Appearance of the Chinese pond mussel in Myanmar requires regular monitoring and other environmental measures from the government and conservation organizations. Dramatic declining of native species in Malaysia as a result of S. woodiana invasion is an example of lacking of special measures for the invasion management (Zieritz et al., 2016). We assume that current local range of S. woodiana in Myanmar provides benefits for invasion control and eradication efforts.

Acknowledgments

We are grateful to Dr. Tony Whitten, Mr. Frank Momberg, Mr. Zau Lunn, and Mr. Nyein Chan (Fauna & Flora International, Myanmar) for their great help during this study. We would like to express our sincerest gratitude to the Department of Fisheries of Ministry of Livestock, Fisheries and Rural Development (Myanmar) and personally to Mr. Myint Than Soe for the permission of the field work and sampling in Myanmar (sampling permission no. 15/6000/MOAI-3103/2016 and export permission no. MOAI/2016-5856).

Declaration of Conflicting Interests

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was partly funded by grants from the President of the Russia Grant Council (no. MD-7660.2016.5), Federal Agency for Scientific Organizations (project nos. 0410-2014-0028 and 0409-2016-0022), Russian Foundation for Basic Research (no. 16-34-00638), and Northern Arctic Federal University.

References

1.

Bolotov I. N. Bespalaya Y. V. Gofarov M. Y. Kondakov A. V. Konopleva E. S. Vikhrev I. V. (2016) Spreading of the Chinese pond mussel, Sinanodonta woodiana, across Wallacea: One or more lineages invade tropical islands and Europe. Biochemical Systematics and Ecology 67: 58–64. https://doi.org/10.1016/j.bse.2016.05.018Google Scholar

2.

Bolotov I. N. Kondakov A. V. Vikhrev I. V. Aksenova O. V. Bespalaya Y. V. Gofarov M. Y. Tumpeesuwan S. (2017a) Ancient river inference explains exceptional oriental freshwater mussel radiations.Scientific Reports 7: 2135. https://doi.org/10.1038/s41598-017-02312-zGoogle Scholar

3.

Bolotov I. N. Vikhrev I. V. Kondakov A. V. Konopleva E. S. Aksenova O. V. Gofarov M. Y. Tumpeesuwan S. (2017b) New taxa of freshwater mussels (Unionidae) from a species-rich but overlooked evolutionary hotspot in Southeast Asia. Scientific Reports 7: article number 11573. https://doi.org/10.1038/s41598-017-11957-9Google Scholar

4.

Djajasasmita, M. (1982). The occurrence of Anodonta woodiana Lea, 1837 in Indonesia (Pelecypoda: Unionidae). Veliger, 25, 175. Google Scholar

6.

Douda K. Velíšek J. Kolářová J. Rylková K. Slavík O. Horký P. Langrová I. (2017) Direct impact of invasive bivalve (Sinanodonta woodiana) parasitism on freshwater fish physiology: Evidence and implications. Biological Invasions 19: 989–999. https://doi.org/10.1007/s10530-016-1319-7Google Scholar

7.

Douda K. Vrtílek M. Slavík O. Reichard M. (2012) The role of host specificity in explaining the invasion success of the freshwater mussel Anodonta woodiana in Europe. Biological Invasions 14(1): 127–137. https://doi.org/10.1007/s10530-011-9989-7Google Scholar

8.

Folmer O. Black M. Hoeh W. Lutz R. Vrijenhoek R. (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3(5): 294. Google Scholar

9.

Froufe E. Lopes-Lima M. Riccardi N. Zaccara S. Vanetti I. Lajtner J. Bogan A. E. (2017) Lifting the curtain on the freshwater mussel diversity of the Italian Peninsula and Croatian Adriatic coast. Biodiversity and Conservation. https://doi.org/10.1007/s10531-017-1403-zGoogle Scholar

10.

Guarneri I. Popa O. P. Gola L. Kamburska L. Lauceri R. Lopes-Lima M. Riccardi N. (2014) A morphometric and genetic comparison of Sinanodonta woodiana (Lea, 1834) populations: does shape really matter?Aquatic Invasions 9: 183–194. https://doi.org/10.3391/ai.2014.9.2.07Google Scholar

11.

Hall T. A. (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98. Google Scholar

12.

Han M. V. Zmasek C. M. (2009) phyloXML: XML for evolutionary biology and comparative genomics. BMC Bioinformatics 10: 356. https://doi.org/10.1186/1471-2105-10-356Google Scholar

13.

Konopleva E. S. Bolotov I. N. Vikhrev I. V. Gofarov M. Y. Kondakov A. V. (2017) An integrative approach underscores the taxonomic status of Lamellidens exolescens, a freshwater mussel from the Oriental tropics (Bivalvia: Unionidae).Systematics and Biodiversity 15(3): 204–217. https://doi.org/10.1080/14772000.2016.1249530Google Scholar

14.

Lajtner J. Crncan P. (2011) Distribution of the invasive bivalve Sinanodonta woodiana (Lea, 1834) in Croatia.Aquatic Invasions 6: S119–S124. https://doi.org/10.3391/ai.2011.6.s1.027Google Scholar

15.

Lopes-Lima M. Sousa R. Geist J. Aldridge D. C. Araujo R. Bergengren J. Zogaris S. (2017) Conservation status of freshwater mussels in Europe: State of the art and future challenges.Biological Reviews 92(1): 572–607. https://doi.org/10.1111/brv.12244Google Scholar

16.

Miller, M., Pfeiffer, W., & Schwartz, T. (2010, November 14). Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Paper presented at the IEEE Gateway Computing Environments Workshop (pp. 1–8). New York, NY: IEEE. Google Scholar

17.

Ng, T. H., Tan, S. K., Wong, W. H., Meier, R., Chan, S. Y., Tan, H. H., & Yeo, D. C. (2016). Molluscs for sale: Assessment of freshwater gastropods and bivalves in the ornamental pet trade. PloS One, 11(8), e0161130 Retrieved from  https://doi.org/10.1371/journal.pone.0161130Google Scholar

18.

Ronquist F. Teslenko M. van der Mark P. Ayres D. L. Darling A. Höhna S. Huelsenbeck J. P. (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61(3): 539–542. https://doi.org/10.1093/sysbio/sys029Google Scholar

19.

Sayenko E. M. Soroka M. Kholin S. K. (2017) Comparison of the species Sinanodonta amurensis Moskvicheva, 1973 and Sinanodonta primorjensis Bogatov et Zatrawkin, 1988 (Bivalvia: Unionidae: Anodontinae) in view of variability of the mitochondrial DNA cox1 gene and conchological features. Biology Bulletin 44(3): 266–276. https://doi.org/10.1134/s1062359017030086Google Scholar

20.

Song X-L. Ouyang S. Zhou C-H Wu X-P. (2016) Complete maternal mitochondrial genome of freshwater mussel Anodonta lucida (Bivalvia: Unionidae: Anodontinae). Mitochondrial DNA A DNA Mapp Seq Anal 27(1): 549–550. https://doi.org/10.3109/19401736.2014.905852Google Scholar

21.

Soroka M. (2005) Genetic variability among freshwater mussel Anodonta woodiana (Lea, 1834) (Bivalvia: Unionidae) populations recently introduced in Poland. Zoological Science 22(10): 1137–1144. https://doi.org/10.2108/zsj.22.1137Google Scholar

22.

Soroka M. (2010a) Characteristics of mitochondrial DNA of unionid bivalves (Mollusca: Bivalvia: Unionidae). I. Detection and characteristics of doubly uniparental in-heritance (DUI) of unionid mitochondrial DNA. Folia Malacol 18: 147–188. https://doi.org/10.2478/v10125-010-0015-yGoogle Scholar

23.

Soroka M. Burzyñski A. (2010b) Complete sequences of maternally inherited mitochondrial genomes in mussels Unio pictorum (Bivalvia, Unionidae). J. Appl. Genet 51: 469–476. https://doi.org/10.1007/bf03208876Google Scholar

24.

Soroka M. Urbańska M. Andrzejewski W. (2014) Chinese pond mussel Sinanodonta woodiana (Lea, 1834) (Bivalvia): origin of the Polish population and GenBank data. J. Limnol 73(3): 454–458. https://doi.org/10.4081/jlimnol.2014.938Google Scholar

25.

Sousa R. Gutiérrez J. L. Aldridge D. C. (2009) Non-indigenous invasive bivalves as ecosystem engineers. Biological Invasions 11: 2367–2385. https://doi.org/10.1007/s10530-009-9422-7Google Scholar

26.

Sousa R. Novais A. Costa R. Strayer D. (2014) Invasive bivalves in fresh waters: Impacts from individuals to ecosystems and possible control strategies. Hydrobiologia 735: 233–251. Retrieved from  http://dx.doi.org/10.1007/s10750-012-1409-1Google Scholar

27.

Tamura K. Stecher G. Peterson D. Filipski A. Kumar S. (2013) MEGA6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30: 2725–2729. https://doi.org/10.1093/molbev/mst197Google Scholar

28.

Thompson J. D. Higgins D. G. Gibson T. J. (1994) CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.Nucleic Acids Research 22: 4673–4680. Google Scholar

29.

Villesen P. (2007) FaBox: An online toolbox for fasta sequences. Molecular Ecology Notes 7: 965–968. https://doi.org/10.1111/j.1471-8286.2007.01821.xGoogle Scholar

30.

Watters G. T. (1997) A synthesis and review of the expanding range of the Asian freshwater mussel Anodonta woodiana (Bivalvia: Unionidae). Veliger 40: 152–156. Google Scholar

31.

Zhang P. Fang H-Y. Pan W-J. Pan H-C. (2016) The complete mitochondrial genome of Chinese pond mussel Sinanodonta woodiana (Unionoida: Unionidae). Mitochondrial DNA A DNA Mapp Seq Anal 27(3): 1620–1621. https://doi.org/10.3109/19401736.2014.958697Google Scholar

32.

Zieritz A. Bogan A. E. Froufe E. Klishko O. Kondo T. Kovitvadhi U. Sousa R. (2017) Diversity, biogeography and conservation of freshwater mussels (Bivalvia: Unionida) in East and Southeast Asia. Hydrobiologia. 1–16. Retrieved from  https://doi.org/10.1007/s10750-017-3104-8Google Scholar

33.

Zieritz A. Lopes-Lima M. Bogan A. E. Sousa R. Walton S. Rahim K. A. A. McGowan S. (2016) Factors driving changes in freshwater mussel (Bivalvia, Unionida) diversity and distribution in Peninsular Malaysia. Science of the Total Environment 571: 1069–1078. https://doi.org/10.1016/j.scitotenv.2016.07.098Google Scholar
© The Author(s) 2017 This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
Ilya V. Vikhrev, Ekaterina S. Konopleva, Mikhail Y. Gofarov, Alexander V. Kondakov, Yulia E. Chapurina, and Ivan N. Bolotov "A Tropical Biodiversity Hotspot Under the New Threat: Discovery and DNA Barcoding of the Invasive Chinese Pond Mussel Sinanodonta Woodiana in Myanmar," Tropical Conservation Science 10(1), (1 January 2020). https://doi.org/10.1177/1940082917738151
Received: 31 July 2017; Accepted: 30 September 2017; Published: 1 January 2020
JOURNAL ARTICLE
PAGES


SHARE
ARTICLE IMPACT
Back to Top