Genetic variation of the Mudskipper (Periophthalmus argentilineatus) in the mangrove ecosystem, North Coast of Aceh, Aceh Province, Indonesia
Article Sidebar
Abstract Views: 336
File Views: 176
Main Article Content
Abstract
Abstract. Dewiyanti I, Siregar PKA, Octavina C, Nurfadillah, Nurfadli, Marimuthu K. 2025. Genetic variation of the Mudskipper (Periophthalmus argentilineatus) in the mangrove ecosystem, North Coast of Aceh, Aceh Province, Indonesia. Biodiversitas 26: 6399-6409. Mudskipper (Periophthalmus argentilineatus), a species belonging to the Gobiidae family of the Perciformes order, is an important ecological indicator of mangrove health, yet information on its genetic diversity in Aceh remains limited. This study analyzes genetic variation in mudskipper populations from the northern coast of Aceh using mitochondrial Cytochrome Oxidase Subunit I (COI) gene sequencing. Although the species has been widely studied in other regions of Indonesia, no molecular genetic baseline has ever been reported for Aceh populations, creating a critical gap in understanding their genetic structure and conservation status. Twenty samples were collected in Aceh Besar and Banda Aceh populations and combined with 19 additional COI sequences obtained from GenBank. The results showed that the mudskipper species in the research locations were P. argentilineatus. In addition, P. argentilineatus populations from Aceh Besar and Banda Aceh showed no detectable genetic variation, as reflected by low genetic distance values indicating close genetic relatedness. This pattern suggests that similar environmental pressure may be contributing to genetic homogeneity between the two populations. However, genetic variation of P. argentilineatus was moderate in the Bali population, and there was a high genetic distance between the Aceh Besar and Bali populations, as well as Banda Aceh and Bali (0.393). These findings highlight the need for conservation strategies that maintain mangrove habitat integrity and promote connectivity among regional mudskipper populations. Maintaining mangrove habitat connectivity and implementing periodic genomic monitoring are critical future steps to preserve gene flow in P. argentilineatus populations along the North Coast of Aceh.
Article Details
Issue
Section
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
References
Aji KW, Arisuryanti T. 2021. Molecular identification of mudskipper fish (Periophthalmus spp.) from Baros Beach, Bantul, Yogyakarta. J Trop Biodivers Biotechnol 6 (3): jtbb66391. DOI: 10.22146/jtbb.66391.
Akbar N, Zamani NP, Madduppa HH. 2014. Keragaman genetik ikan tuna sirip kuning (Thunnus albacares) dari dua populasi di Laut Maluku, Indonesia. Depik 3 (1): 65-73. DOI: 10.13170/depik.3.1.1304. [Indonesia]
Ali FS, Ismail M, Aly W. 2020. DNA barcoding to characterize the biodiversity of freshwater fishes of Egypt. Mol Biol Rep 47: 5865-5877. DOI: 10.1007/s11033-020-05657-3.
Allio R, Donega S, Galtier N, Nabholz B. 2017. Large variation in the ratio of mitochondrial to nuclear mutation rate across animals: Implications for genetic diversity and the use of mitochondrial DNA as a molecular marker. Mol Biol Evol 34 (11): 2762-2772. DOI: 10.1093/molbev/msx197.
Ansari A, Trivedi S, Saggu S, Rehman H. 2014. Mudskipper: A biological indicator for environmental monitoring and assessment of coastal waters. J Entomol Zool Stud 2 (6): 22-33. DOI:10.1111/j.1095-. 8649.2010.02807.
Arisuryanti T, Hasan RL, Koentjan JP. 2018. Genetic identification of two mudskipper species (Pisces: Gobiidae) from Bogowonto Lagoon (Yogyakarta, Indonesia) using COI mitochondrial gene as a DNA barcoding marker. AIP Conf Proc 2002 (1): 020068. DOI: 10.1063/1.5050164.
Arisuryanti T, Aji KW, Shabrina FN, Febriyanti D, Daryono BS, Priyono DS. 2024a. Phylogenetic and genetic variation of common mudskippers (Periophthalmus kalolo Lesson, 1831) from the southern coast of Java, Indonesia, inferred from the COI mitochondrial gene. J Genet Eng Biotechnol 22 (1): 100335. DOI: 10.1016/j.jgeb.2023.100335.
Arisuryanti T, Aji KW, Herawati H, Sari IP, Rha’ifa1 FA, Febriyanti D, Priyono DS. 2024b. Cryptic diversity of barred mudskippers, Periophthalmus argentilineatus (Valenciennes, 1837), from the Southern Coast of Java and East Lombok, Indonesia, inferred by COI Mitochondrial Gene. J Trop Biodivers Biotechnol 9 (3): jtbb84328. DOI: 10.22146/jtbb.84328.
Assal N, Lin M. 2021. PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis. J Microbiol Methods 181: 106121. DOI: 10.1016/j.mimet.2020.106121.
Baderan DWK, Aydalina RV, Hamidun MS. 2023. Morphological characteristics and biodiversity of mudskipper fish (Periophthalmus: Gobiidae) in mangrove ecosystem of coastal Bay of Tomini, Boalemo, Gorontalo Province, Indonesia. Biodiversitas 24 (1): 498-507. DOI: 0.13057/biodiv/d240158.
Basith A, Abinawanto A, Kusrini E, Yasman Y. 2021. Genetic diversity analysis and phylogenetic reconstruction of groupers Epinephelus spp. from Madura Island, Indonesia, based on partial sequence of CO1 gene. Biodiversitas 22: 4282-4290. DOI: 10.13057/biodiv/d221020.
Bhattacharya M, Sharma AR, Patra BC, Sharma G, Seo EM, Nam JS, Chakraborty C, Lee SS. 2016. DNA barcoding to fishes: Current status and future directions. Mitochondrial DNA A DNA Mapp Seq Anal 27 (4): 2744-2752. DOI: 10.3109/19401736.2015.1046175.
Bidawi BM, Desrita D, Yunasfi Y. 2017. Hubungan panjang berat dan faktor kondisi ikan belodok (Famili: Gobiidae) pada ekosistem mangrove di Desa Pulau Sembilan Kabupaten Langkat Provinsi Sumatera Utara. Depik 6 (3): 228-234. DOI: 10.13170/depik.6.3.7029. [Indonesia].
Bilandžija H, Morton B, Podnar M, Ćetković H. 2013. Evolutionary history of relict Congeria (Bivalvia: Dreissenidae): Unearthing the subterranean biodiversity of the Dinaric Karst. Front Zool 10 (5): 1-8. DOI: 10.1186/1742-9994-10-5.
Binashikhbubkr K, Al-Misned F, Naim DM. 2024. Genetic diversity and population structure of Kawakawa Euthynnus affinis (Cantor, 1849) in Malaysia and its surrounding waters inferred by mitochondrial DNA cytochrome oxidase I and cytochrome b genes. J King Saud Univ Sci 36: 103193. DOI: 10.1016/j.jksus.2024.103193.
Bolaji DA, Aderonke OLA, Minasu PK. 2023. DNA barcoding and misidentification of some marine fish species in Nigerian industrial trawl fishery. Sci Afr 20: e01662. DOI: 10.1016/j.sciaf.2023.e01662.
Chan BK, Tsao YF, Wangkulangkul K, Amjud K, Sukparangsi W. 2022. Biogeography and biodiversity of the intertidal barnacle Tetraclita species in the Gulf of Thailand and Andaman Sea-influences oceanographic currents and Pleistocene glaciations. Front Mar Sci 8: 774041. DOI: 10.3389/fmars.2021.774041.
Cheng J, Kao H, Dong S. 2020. Population genetic structure and gene flow of rare and endangered Tetraena mongolica Maxim. revealed by reduced representation sequencing. BMC Plant Biol 20: 391. DOI: 10.1186/s12870-020-02594-y.
Dahruddin H, Hutama A, Busson F, Sauri S, Hanner R, Keith P, Had-iaty RK, Hubert N. 2017. Revisiting the ichthyo-diversity of Java and Bali through DNA barcodes: Taxonomic coverage, identification accuracy, cryptic diversity, and identification of exotic species. Mol Ecol Resour 17 (2): 288-299. DOI: 10.1111/1755-0998.12528.
Dewiyanti I, Melanie K, Almuniro S, Damora A, Nufadillah N, Batubara, AS. 2022. Growth patterns and condition factor of the mudskipper (Periophthalmus gracilis) in mangrove ecosystem rehabilitation areas in Banda Aceh and Aceh Besar, Indonesia. Fish Aquat Life 30: 85-94. DOI: 10.2478/aopf-2022-0008.
Dewiyanti I, Rifki M, Octavina C, Ulfa M, Damora A, Nurfadillah N. 2023. Important value index (IVI) and diversity of mangrove vegetation in Aceh Tamiang, Aceh Province. Depik 12 (2): 190-197. DOI: 10.13170/depik.12.2.31130.
Dias PJ, Gilg MR, Lukehurst SS, Kennington WJ, Huhn M, Madduppa HH, McKirdy SJ, De Lestang P, Teo SL, Lee SS, McDonald JI. 2018. Genetic diversity of a hitchhiker and prized food source in the Anthropocene: The Asian green mussel Perna viridis (Mollusca, Mytilidae). Biol Invasions 20: 1749-1770. DOI: 10.1007/s10530-018-1659-6.
Freudiger A, Josi D, Thünken T, Herder F, Flury JM, Marques DA, Taborsky M, Frommen JG. 2021. Ecological variation drives morphological differentiation in a highly social vertebrate. Funct Ecol 35 (10): 2266-2281. DOI: 10.1111/1365-2435.13857.
Grewe PM, Krueger CC, Aquadro CF, Bermingham E, Kincaid HL, May B. 1993. Mitochondrial DNA variation among lake trout (Salvelinus namaycush) strains stocked into Lake Ontario. Can J Fish Aquat Sci 50 (11): 2397-2403. DOI:10.1139/f93-264.
Habib KA, Neogi AK, Rahman M, Oh J, Lee YH, Kim CG. 2021. DNA barcoding of brackish and marine water fishes and shellfish of Sundarbans, the world's largest mangrove ecosystem. PLoS One 16 (8): e0255110. DOI: 10.1371/journal.pone.0255110.
Hakim AA, Dikrurahman, Sahidan M, Retno PA, Dyah WS, Mohammad MK. 2022. A genetic approach for authentication on morphological differences of cultivated pompano species in Indonesia. Biodiversitas 23 (9): 4561-4569. DOI: 10.13057/biodiv/d230923.
Hebert PD, Cywinska A, Ball SL, DeWaard JR. 2003. Biological identifications through DNA barcodes. Biol Sci 270: 313-321. DOI: 10.1098/rspb.2002.2218.
Hedrick PW. 2005. A standardized genetic differentiation measure. Evolution 59: 1633-1638. DOI: 10.1111/j.0014-3820. 2005.tb01814.x.
Hefzi F, Mansyurdin M, Tesri M, Nurainas N. 2023. Genetic variation of Ceiba pentandra (L.) Gaertn. from three populations in West Sumatra, Indonesia, based on RAPD Markers. Intl J Prog Sci Technol 40 (1): 116-126. DOI: 10.52155/ijpsat.v40.1.5507.
Hillis DM, Bull JJ. 1993. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Syst Biol 42 (2): 182-192. DOI: 10.1093/sysbio/42.2.182.
Hohenlohe PA, Funk WC, Rajora OP. 2021. Population genomics for wildlife conservation and management. Mol Ecol 30 (1): 62-82. DOI: 10.1111/mec.15720.
Imtiaz A, Mohd-Nor SA, Darlina MDN. 2017. Review: Progress and potential of DNA barcoding for species identification of fish species. Biodiversitas 18 (4): 1394-1405. DOI: 10.13057/biodiv/d180415.
Jaafar Z, Perrig M, Ming-Chou L. 2009. Periophthalmus variabilis (Teleostei: Gobiidae: Oxudercinae), a valid species of mudskipper, and a re-diagnosis of Periophthalmus novemradiatus. Zool Sci 26 (4): 309-314. DOI: 10.2108/zsj.26.309.
Jimenez LCZ, Queiroz HM, Otero XL, Nóbrega GN, Ferreira TO. 2021. Soil organic matter responses to mangrove restoration: A replanting experience in Northeast Brazil. Intl J Environ Res Public Health 18 (17): 8981. DOI: 10.3390/ijerph18178981.
Kalendar R, Boronnikova S, Seppänen M. 2021. Isolation and purification of DNA from complicated biological samples. Methods Mol Biol 2222: 57-67. DOI: 10.1007/978-1-0716-0997-2_3.
Khusna UA, Fadzilah HN, Raihan M, Pramesti HP, Salfiah EN, Rahayu DA, Nugroho ED, Rahim AA, Rusdianto. 2024. DNA barcode of mudskipper based on DNA mini-barcode COI from the North Coast of East Java, Indonesia. AACL Bioflux 17 (6): 3063-3075.
Kusuma AB, Bengen DG, Madduppa H, Subhan B, Arafat D. 2016. Keanekaragaman genetik karang lunak Sarcophyton trocheliophorumpada populasi Laut Jawa, Nusa Tenggara dan Sulawesi. Enggano 1 (1): 89-96. DOI: 10.31186/jenggano.1.1.89-96. [Indonesia]
Lee PY, Costumbrado J, Hsu CY, Kim YH. 2012. Agarose gel electrophoresis for the separation of DNA fragments. J Vis Exp 20 (62): e3923. DOI: 10.3791/3923.
Leitwein M, Duranton M, Rougemont Q, Gagnaire PA, Bernatchez L. 2020. Using haplotype information for conservation genomics. Trends Ecol Evol 35 (3): 245-258. DOI: 10.1016/j.tree.2019.10.012.
Li J, Ye Y, Wu C, Qi P, Guo B, Chen Y. 2013. Genetic variation of Mytilus coruscus Gould (Bivalvia: Mytilidae) populations in the East China Sea inferred from mtDNA COI gene Sequence. Biochem Syst Ecol 50: 30-38. DOI: 10.1016/j.bse.2013.03.033.
Li FW, Kuo LY, Pryer KM, Rothfels CJ. 2016. Genes translocated into the plastid inverted repeat show decelerated substitution rates and elevated GC content. Genome Biol Evol 8 (8): 2452-2458. DOI: 10.1093/gbe/evw167.
Librado P, Rozas J. 2009. DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25 (11): 1451-1452. DOI: 10.1093/bioinformatics/btp187.
Luo YJ, Satoh N, Endo K. 2015. Mitochondrial gene order variation in the brachiopod Lingula anatina and its implications for mitochondrial evolution in lophotrochozoans. Mar Genom 24 (1): 31-40. DOI: 10.1016/j.margen.2015.08.005.
Marnis H, Syahputra K, Darmawan J, Febrianti D, Tahapari E, Larashati S, Iswanto B, Primanita EHP, Syaifudin M, Subangkit AT. 2024. DNA barcoding of fish diversity from Batanghari River, Jambi, Indonesia. Fish Aquat Sci 27: 87-99. DOI: 10.47853/FAS.2024.e10.
Muhtadi A, Ramadhani S, Yunasfi. 2016. Identification and habitat type of mudskipper (Family: Gobiidae) at the Bali Beach, District of Batu Bara, North Sumatra Province. Biospecies 9 (2): 1-6. DOI: 10.22437/Biospecies.9.2.3156.
Naresh S, Kunasundari B, Gunny AAN, Teoh YP, Shuit SH, Ng QH, Hoo PY. 2019. Isolation and partial characterization of thermophilic cellulolytic bacteria from north Malaysian tropical mangrove soil. Trop Life Sci Res 30 (1): 123-147. DOI: 10.21315/tlsr2019.30.1.8.
Natsir M, Ulya Z, Fitriani R. 2022. Mangrove forest utilization policies reconceptualized with a view to improving the regional economy in Aceh Tamiang District, Indonesia. Biodiversitas 23 (12): 6570-6578. DOI: 10.13057/biodiv/d231256.
Nelson JS, Grande TC, Wilson MV. 2016. Fishes of the World. John Wiley and Sons, United States of America.
Octavina C, Ramadhaniaty M, Daulay RE, Dewiyanti I, Ulfah M. Genetic variation and population structure of brachiopods, Lingula anatina Lamarck, 1801 in the Northern Aceh Shore, Indonesia. Biodiversitas 24 (7): 3951-3959. DOI: 10.13057/biodiv/d240733.
Oh A, Oh-Un B. 2022. Genetic differentiation that is exceptionally high and unexpectedly sensitive to geographic distance in the absence of gene flow: Insights from the genus Eranthis in East Asian regions. Ecol Evol 12: e9007. DOI: 10.1002/ece3.9007.
Osman AGM, El-Ganainy A, Abd-Allah E. 2018. Some reproductive aspects of the areolate grouper, Epinephelus areolotus, from the Gulf of Suez. Egypt J Aquat Res 44 (1): 51-56. DOI: 10.1016/j.ejar.2018.02.002.
Pino‐Querido A, Álvarez‐Castro JM, Vera M, Pardo BG, Fuentes J, Martínez P. 2015. A molecular tool for parentage analysis in the Mediterranean mussel (M. galloprovincialis). Aquac Res 46: 1721-1735. DOI: 10.1111/are.12329.
Polgar G, Zane L, Babbucci M, Barbisan F, Patarnello T, Rüber L, Papetti C. 2024. Phylogeography and demographic history of two widespread Indo-Pacific mudskippers (Gobiidae: Periophthalmus). Mol Phylogenet Evol 73: 161-176. DOI: 10.1016/j.ympev.2014.01.014.
Quang-Le H, Suffredini E, Tien-Pham D, Kim TA, De-Medici D. 2018. Development of a method for direct extraction of viral RNA from bivalve molluscs. Lett Appl Microbiol 67: 426-434. DOI: 10.1111/lam.13065.
Raharinaivo LR, Jaonalison H, Mahafina J, Ponton D. 2020. How to efficiently determine the size at maturity of small-sized tropical fishes: A case study based on 144 species identified via DNA barcoding from southwestern Madagascar. J Appl Ichthyol 36: 402-413. DOI: 10.1111/jai.14046.
Ramadhaniaty M, Setyobudiandi I, Madduppa H. 2018. Morphogenetic and population structure of two species of marine bivalve (Ostreidae: Saccostrea cucullata and Crassostrea iredalei) in Aceh, Indonesia. Biodiversitas 19 (3): 978-988. DOI: 10.13057/biodiv/d190329.
Rebman JP, Simpson MG. 2006. Checklist of the Vascular Plants of San Diego County, 5th edition. San Diego Natural History Museum, San Diego, California.
Rha'ifa FA, Audrea DJ, Hakim L, Arisuryanti T. 2021. DNA barcode of barred mudskipper (Periophthalmus argentilineatus Valenciennes, 1837) from the Tekolok estuary (West Nusa Tenggara, Indonesia) and their phylogenetic relationship with other Indonesian barred mudskippers. J Trop Biodivers Biotechnol 6 (2): 1-13. DOI: 10.22146/jtbb.59702.
Rizal S, Damm P, Wahid MA, Sundermann J, Ilhamsyah Y, Iskandar T, Muhammad M. 2012. General circulation in the Malacca Strait and Andaman Sea: A numerical model study. Am J Environ Sci 8 (5): 479-488. DOI: 10.3844/ajessp.2012.479.488.
Rozas J, Ferrer-Mata A, Sánchez-DelBarrio JC, Guirao-Rico S, Librado P, Ramos-Onsins SE, Sánchez-Gracia A. 2017. DnaSP 6: DNA sequence polymorphism analysis of large data sets. Mol Biol Evol 34 (12): 3299-3302. DOI: 10.1093/molbev/msx248.
Saleky D, Leatemia SP, Pattiasina TF, Isma I, Pangaribuan RD, Welliken MA, Melmambessy EH, Dailami M. 2020. Analisis pola pertumbuhan dan pendekatan DNA barcoding untuk identifikasi Turbo stenogyrus P. Fischer, 1873 (Mollusca: Gastropoda). Biotropika 8 (2): 79-86. DOI: 10.21776/ub.biotropika. [Indonesia]
Saleky D, Dailami M. 2021. Konservasi genetik ikan kakap putih (Lates calcarifer, Bloch, 1790) melalui pendekatan DNA barcoding dan analisis filogenetik di sungai Kumbe Merauke Papua. Jurnal Kelautan Tropis 24 (2): 141-150. DOI: 10.14710/jkt.v24i2.10760. [Indonesia]
Sambrook J, Russell DW. 2001. Molecular Cloning: Ch. 8. In Vitro Amplification of DNA by the Polymerase Chain Reaction Vol. 2. Cold Spring Harbor Laboratory Press, Michigan University, United States.
Santoso H, Suhartono E, Yunita R, Biyatmoko D, Banjarbaru U. 2020. Mudskipper fish as a bio-indicator for heavy metals pollution in a Coastal Wetland. Egypt J Aquat Biol Fish 24 (7): 1073-1095. DOI: 10.21608/ejabf.2020.144402.
Schreiber HA, Bitner MA, Carlson SJ. 2013. Morphological analysis of phylogenetic relationships among extant rhynchonellide brachiopods. J Paleontol 87 (4): 550-569. DOI: 10.1666/12-115.
Shabrina FN, Wibowo K, Arisuryanti T. 2024. Cryptic diversity of mudskipper genus Boleophthalmus (Gobiiformes: Oxudercidae) from the north coast of East Java, Indonesia. Biodiversitas 25 (1): 412-420. DOI: 10.13057/biodiv/d250148.
Shipilina D, Pal A, Stankowski S, Chan YF, Barton NH. 2023. On the origin and structure of haplotype blocks. Mol Ecol 32: 1441-1457. DOI: 10.1111/mec.16793.
Sokefun O, Gan HM, Tan MP. 2022. Characterization of the Nigerian mudskipper species: Phylogenetic relationship, population structure, and DNA barcoding using the Cytochrome Oxidase 1 (CO1) gene. Intl J Fauna Biol Stud 9: 48-51. DOI: 10.22271/23940522.2022.v9.i4a.923.
Soltis PS, Douglas ES. 2003. Applying the bootstrap in phylogeny reconstruction. Statist Sci 18: 256-267. DOI: 10.1214/ss/10639949802003.
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 30 (12): 2725-2729. DOI: 10.1093/molbev/mst197.
Temereva EN, Gebruk AA, Malakhov VV. 2015. Demonstration of the preoral coelom in the brachiopod Lingula anatina with consideration of its phylogenetic significance. Zool Anzeiger-A J Comp Zool 256: 22-27. DOI: 10.1016/j.jcz.2015.03.002.
Tresnati J, Aprianto R, Tuwo A. 2021. Ecosystem and Sustainable Use of Mangrove Forests. Integrated Publications, New Delhi.
Triandiza T, Madduppa H. 2018. Aplikasi analisa morfologi dan DNA barcoding pada penentuan jenis kepiting porcelain (Pisidia sp.) yang berasal dari Pulau Tunda, Banten. Jurnal Sumberdaya Akuatik Indopasifik 2 (2): 81-90. DOI: 10.30862/jsai.v2i2.51. [Indonesia]
Walker SE, Lorsch J. 2013. Sanger dideoxy sequencing of DNA. Methods Enzymol 529: 171-184. DOI: 10.1016/B978-0-12-418687-3.00014-8.
Wang T, Zhang YP, Yang ZY, Liu Z, Du YY. 2020. DNA barcoding reveals cryptic diversity in the underestimated genus Triplophysa (Cypriniformes: Cobitidae, Nemacheilinae) from the northeastern Qinghai-Tibet Plateau. BMC Evol Biol 20 (1): 151. DOI: 10.1186/s12862-020-01718-0.
Wright S. 1978. Evolution and the Genetics of Population, Variability within and Among Natural Populations. The University of Chicago Press, Chicago.
Xin Q, Qing J, He Y. 2024. Analysis of kinship and population genetic structure of 53 apricot resources based on whole genome resequencing. Curr Issues Mol Biol 46 (12): 14106-14118. DOI: 10.3390/cimb46120844.
Ye Y, Ren W, Zhang S, Zhao L, Tang J, Hu L, Chen X. 2022. Genetic diversity of fish in aquaculture and of common carp (Cyprinus carpio) in traditional rice-fish coculture. Agriculture 12 (7): 997. DOI: 10.3390/ agriculture12070997.
Zizka VMA, Weiss M, Leese F. 2020. Can metabarcoding resolve intraspecific genetic diversity changes to environmental stressors? A test case using river macrozoobenthos. Metabarcod Metagenom 4: 23-34. DOI: 10.3897/mbmg.4.51925.