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DNA Barcoding of Freshwater Eels Anguilla spp from Kuari River Based on Cytochrome C Oxidase Subunit I (COI) Gene
Corresponding Author(s) : Mochamad Syaifudin
Jurnal Ilmiah Perikanan dan Kelautan, 2024: IN PRESS ISSUE (JUST ACCEPTED MANUSCRIPT, 2024)
Abstract
Graphical Abstract
Highlight Research
- Freshwater eels in Kuari River consisted of two species marmorata and A. bengalensis
- Anguilla marmorata had the highest similarity of 99.82%-100%, while bengalensis indicated the highest identity of 99.84%-100% to the same species in the GenBank database.
- Phylogenetic species indicated that Anguilla marmorata and Anguilla bengalensis form two different sub-clusters.
- The water quality in the Bengkulu Kuari River were still in tolerance for fish growth and survival.
Abstract
Freshwater eels (Anguilla spp.) are classified in the family Anguillidae, and included in the catadromous group. This study aimed to determine the COI gene sequences of the mitochondrial DNA, analyze the genetic distances, and phylogenetics, and characterize the physical and chemical parameters of between freshwater eels habitat in the Kuari River Bengkulu. This research was conducted from November 2020 – April 2021. The methods used in barcoding eel species were DNA isolation, DNA amplification using PCR (Polymerase Chain Reaction), electrophoresis, and sequencing of COI gene regions in mtDNA. The COI mtDNA gene fragments were obtained from PCR results with an annealing temperature of 50°C for 30 seconds in 35 cycles. BLASTn analysis of eel samples AM3 and AM4 had the highest similarity of 99.82%-100% to Anguilla marmorata, and samples AB2-AB5 indicated the highest identity of 99.84%-100% to Anguilla bengalensis. Phylogenetic species indicated that Anguilla marmorata and Anguilla bengalensis form two different sub-clusters. The water qualities in the Bengkulu Kuari River were temperatures 26.5-27.5oC, pH 7.1-8.7, dissolved oxygen 6.19-9.54 mg L-1, brightness 21-47 cm, ammonia 0.16-0.41 mg L-1, total alkalinity 20–52 mg L-1, TDS 33–49 mg L-1, salinity 0.3-0.4 ppt and water velocity 0.5-0.8 ms-1. The COI gene in DNA barcoding is very appropriate to be used for the identification of Anguilla spp species by comparing the DNA sequence of the COI gene with the existing database in the Genbank.
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- Affandi, R. (2005). Strategy on utilization of eel (Anguilla sp.) resources in Indonesia. Jurnal Iktiologi Indonesia, 5(2):77-81.
- Anjarsari, Y., Syaifudin, M., Jubaedah, D., Taqwa, F. H., & Yonarta, D. (2021). Phylogenetic of featherback Chitala sp from South Sumatra based on cytochrome C oxidase subunit I (COI) gene. IOP Conference Series: Earth and Environmental Science, 810(2):1-7.
- Arai, T., Chin, T. C., Kwong, K. O., & Azizah, M. N. S. (2015). Occurrence of the tropical eels Anguilla bengalensis bengalensis and Anguilla bicolor bicolor in Peninsular Malaysia and implications for eel taxonomy. Marine Biodiversity Records, 8(28):1-4.
- Arai, T., & Taha, H. (2021). Contrasting patterns of genetic population structure in tropical freshwater eels of genus Anguilla in the Indo-Pacific. Heliyon, 7(5):1-10.
- Arai, T., Chai, I. J., Iizuka, Y., & Chang, C. W. (2020). Habitat segregation and migration in tropical anguillid eels, Anguilla bengalensis bengalensis and A. bicolor bicolor. Scientific Reports, 10(1):1-13.
- Arai, T., & Chino, N. (2018). Opportunistic migration and habitat use of the giant mottled eel Anguilla marmorata (Teleostei: Elopomorpha). Scientific Reports, 8(1):1-10.
- Avise, J. C. (1989). Gene trees and organismal histories: A phylogenetic approach to population biology . Evolution, 43(6):1192-1208.
- Barendse, J., Roel, A., Longo, C., Andriessen, L., Webster, L. M. I., Ogden, R., & Neat, F. (2019). DNA barcoding validates species labelling of certified seafood. Current Biology 29(6):198-199.
- Basith, A., Kusrini, E., Abinawanto, A., & 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(10):4282-4290.
- Bengkulu Provincial Government. (2021). Bengkulu at a glance.
- Briones, A. A., Yambot, A. V, Shiao, J.-C., Iizuka, Y., & Tzeng, W.-N. (2007). Migratory pattern and habitat use of tropical eels Anguilla spp. (Teleostei: Anguilliformes: Anguillidae) in the Philippines, as revealed by otolith microchemistry. The Raffles Bulletin of Zoology 14(1):141-149.
- European Food Safety Authority (EFSA). (2009). Scientific opinion: Animal welfare aspects of husbandry systems for farmed European eel. The European Food Safety Authority Journal, 809(10):1-18.
- Fahmi, M. R. (2015). Conservation of genetic tropical fish eel (Anguilla spp) waters in Indonesia. [in English]. Jurnal Penelitian Perikanan Indonesia, 21(1):45-54.
- Falah, I. N., Adharini, R. I., & Ratnawati, S. E. (2023). Molecular identification of elvers (Anguilla spp.) from river estuaries in Central Java, Indonesia using DNA barcoding based on mtDNA CO1 sequences. Jurnal Ilmiah Perikanan dan Kelautan, 15(1):121-130.
- Fekri, L., Affandi, R., Rahardjo, M. F., Budiardi, T., Simanjuntak C. P. H., Fauzan, T. & Indrayani, I. (2018). The effect of temperature on the physiological condition and growth performance of freshwater eel elver Anguilla bicolor bicolor (McClelland, 1844). Jurnal Akuakultur Indonesia, 17(2):181-190.
- Fishbase. (2021, June 12). Anguilla bengalensis and Anguilla marmorata.
- Government Regulation of Republic Indonesia No. 82. (2001). Water quality management and control water pollution. Jakarta.
- Han, Y-S, Yambot, A. V, Zhang, H, & Hung, C-L. (2012). Sympatric spawning but allopatric distribution of Anguilla japonica and Anguilla marmorata: Temperature- and oceanic current-dependent sieving. Plos One, 7(6):1-11.
- Han, C., Dong, S., Li, L., Gao, Q., & Zhou, Y. (2021). DNA barcoding and mini-DNA barcoding reveal mislabeling of salmonids in different distribution channels in the Qingdao area. Journal of Ocean University of China, 20(6):1537-1544.
- Hanzen, C., Lucas, M. C., O’Brien, G., Downs, C. T., & Willows-Munro, S. (2020). African freshwater eel species (Anguilla spp.) identification through DNA barcoding. Marine and Freshwater Research, 71(11):1543-1548.
- Hebert, P. D. N., Ratnasingham, S., & deWaard, J. R. (2003). Barcoding animal life: Cytochrome C oxidase subunit 1 divergences among closely related species. Proceedings. Biological Sciences / The Royal Society, 270(1):96-99.
- Hillis, D. M., & Bull, J. J. (1993). An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology, 42(2):182-192.
- Huyen, K. T., & Linh, N. Q. (2020). Phylogenetic analysis of Anguilla marmorata population in Thua Thien Hue, Vietnam based on the cytochrome C oxidase I (COI) gene fragments. AMB Express, 10(1):1-8.
- Jellyman, D. J, & Arai T. (2016). Juvenile eels: Upstream migration and habitat use. In: T. Arai (Ed.), Biology and ecology of Anguillid eels. (pp. 143-170) CRC Press Taylor & Francis Group.
- Kardin, Sara, L., & Pangerang, U. K. (2016). Some biological aspects of eels (Anguilla sp.) in Mosolo Waters of Wawonii Island, Konawe Archipelago. Manajemen Sumberdaya Perairan, 1(4):355-365.
- Kultz, D. (2005). Molecular and evolutionary basis of the cellular stress response. Annual Review Physiology, 2005(67):225-257.
- Laudien, J., Flint, N. S., Van Der Bank, F. H., & Brey, T. (2003). Genetic and morphological variation in four populations of the surf clam Donax serra (Röding) from Southern African sandy beaches. Biochemical Systematics and Ecology, 31(7):751-772.
- Lawson T. B. (1995). Fundamentals of aquacultural engineering. New York: Chapmanand Hall.
- Mallya, Y. J. (2007). The effects of dissolved oxygen on fish growth in aquaculture. UNU-Fisheries Training Programme. 30 pp.
- Marnis, H., Syahputra, K., Darmawan, J., Febrianti, D., Tahapari, E., Larashati, S., Iswanto, B., Primanita, E. P. H., Syaifudin, M., & Subangkit, A. T. (2024). DNA barcoding of fish diversity from Batanghari River, Jambi, Indonesia. Fisheries and Aquatic Sciences, 27(2):87-99.
- Muchlisin, Z. A., Batubara, A. S., Fadli, N., Muhammadar, A. A., Utami, A. I., Farhana, N., & Siti-Azizah, M. N. (2017). Assessing the species composition of tropical eels (Anguillidae) in Aceh Waters, Indonesia, with DNA barcoding gene cox1. F1000Research, 6(258):1-11.
- Mulis. (2015). Enlargement of eel seeds with different types of feed. Scientific Journal of Fisheries and Marine Affairs, 3(1):20-24.
- Peninal, S., Subramanian, J., Elavarasi, A., & Kalaiselvam, M. (2017). Genetic identification of marine eels through DNA barcoding from Parangipettai coastal waters. Genomics Data, 11(1):81-84.
- Putra, G. I. N., Indrawan, G. S., & Faiqoh, E. (2024). DNA barcoding of cardinalfish (Apogonidae) in Gilimanuk Bay, Bali, Indonesia. Jurnal Ilmiah Perikanan dan Kelautan, 16(1):1-10.
- Rasmussen, R. S., Morrissey, M. T., & Hebert, P. D. N. (2009). DNA barcoding of commercially important salmon and trout species (Oncorhynchus and Salmo) from North America. Journal of Agricultural and Food Chemistry, 57(18):8379-8385.
- Rasmussen, M.D. & Kellis, M. (2011). Accurate gene-tree reconstruction by learning gene-and species-specific substitution rates across multiple 59 complete genomes. Genome Research, 17(12):1932-1942.
- Saanin, H. (1984). Taxonomy and key identification of fish. Volumes 1 and 2. Jakarta: Bina Cipta.
- Sadler, K. (1981). The toxicity of ammonia to the European eel (Anguilla anguilla L.). Aquaculture, 26(1-2):173-181.
- Silfvergrip, A. M. (2009). Report 5943 - CITES identification guide to the freshwater eels (Anguillidae) with focus on the European eel Anguilla anguilla.
- Stecher, G., Knyaz, C., Tamura, K., & Kumar, S. (2020). Molecular evolutionary genetics analysis (MEGA) for macOS. Molecular Biology and Evolution, 37(4):1237-1239.
- Syaifudin, M., Bekaert, M., Taggart, J., Hulata, G., D’Cotta, H., Baroiller, J., Penman, D., & McAndrew, B. (2017a). DNA typing across ten tilapia species using cytochrome C oxidase subunit I (COI) in Abstracts: Twelfth International Symposium on Genetics in Aquaculture 2015.. Aquaculture, 472(4/1):125-125.
- Syaifudin, M., Gultom, E. T., & Wijayanti, M. (2023). DNA authentication of Indonesian leaffish Pristolepis grooti from Kelekar River and Ogan River in South Sumatra based on cytochrome C oxidase subunit I (COI) Gene. Journal of Tropical Biodiversity and Biotechnology, 8(2):1-11.
- Syaifudin, M., Jubaedah, D., Muslim, M., & Daryani, A. (2017b). DNA authentication of Asian redtail catfish Hemibagrus nemurus from Musi and Penukal River, South Sumatra Indonesia. Genetics of Aquatic Organisms, 1(12):43-48.
- Syaifudin, M., Jubaedah, D., Taqwa, F. H., & Octaviani, R. (2021). Phylogenetic of marble goby (Oxyeleotris marmorata Blkr.) in South Sumatra based on cytochrome C oxidase subunit I (COI) gene. Genetics of Aquatic Organisms, 6(1):1-6.
- Tamura, K., Stecher, G., & Kumar, S. (2021). MEGA1: Molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution, 38(7):3022-3027.
- Ward, R. D., Zemlak, T. S, Innes, B. H, Last, P. R., Hebert, P. D. N. (2005). DNA barcoding Australia’s fish species. Philosophical Transactions of the Royal Society B, 360(1462):1847–57.
- Watanabe, S., Aoyama, J., Miller, M. J., Ishikawa, S., Feunteun, E., & Tsukamoto, K. (2008). Evidence of population structure in the giant mottled eel, Anguilla marmorata, using total number of vertebrae. Copeia, 2008(3):680-688.
- Wibowo, A., Hubert, N., Dahruddin, H., Steinke, D., Suhaimi, R. A., Samuel, Atminarso, D., Anggraeni, D. P., Trismawanti, I., Baumgartner, L. J., & Ning, N. (2021). Assessing temporal patterns and species composition of glass eel (Anguilla spp.) cohorts in Sumatra and Java using DNA barcodes. Diversity, 13(5):1-15.
- Zan, N. D., Sarbini, A., Taha, H., Tan, I. V., Azri, A., Kahar, R., Metali, F., Ahmad, N., & Arai, T. (2020). Occurrence and ecological implication of a tropical anguillid eel, Anguilla marmorata, in Brunei Darussalam, Borneo Island. Zoologia, 37(6):1-7.
References
Affandi, R. (2005). Strategy on utilization of eel (Anguilla sp.) resources in Indonesia. Jurnal Iktiologi Indonesia, 5(2):77-81.
Anjarsari, Y., Syaifudin, M., Jubaedah, D., Taqwa, F. H., & Yonarta, D. (2021). Phylogenetic of featherback Chitala sp from South Sumatra based on cytochrome C oxidase subunit I (COI) gene. IOP Conference Series: Earth and Environmental Science, 810(2):1-7.
Arai, T., Chin, T. C., Kwong, K. O., & Azizah, M. N. S. (2015). Occurrence of the tropical eels Anguilla bengalensis bengalensis and Anguilla bicolor bicolor in Peninsular Malaysia and implications for eel taxonomy. Marine Biodiversity Records, 8(28):1-4.
Arai, T., & Taha, H. (2021). Contrasting patterns of genetic population structure in tropical freshwater eels of genus Anguilla in the Indo-Pacific. Heliyon, 7(5):1-10.
Arai, T., Chai, I. J., Iizuka, Y., & Chang, C. W. (2020). Habitat segregation and migration in tropical anguillid eels, Anguilla bengalensis bengalensis and A. bicolor bicolor. Scientific Reports, 10(1):1-13.
Arai, T., & Chino, N. (2018). Opportunistic migration and habitat use of the giant mottled eel Anguilla marmorata (Teleostei: Elopomorpha). Scientific Reports, 8(1):1-10.
Avise, J. C. (1989). Gene trees and organismal histories: A phylogenetic approach to population biology . Evolution, 43(6):1192-1208.
Barendse, J., Roel, A., Longo, C., Andriessen, L., Webster, L. M. I., Ogden, R., & Neat, F. (2019). DNA barcoding validates species labelling of certified seafood. Current Biology 29(6):198-199.
Basith, A., Kusrini, E., Abinawanto, A., & 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(10):4282-4290.
Bengkulu Provincial Government. (2021). Bengkulu at a glance.
Briones, A. A., Yambot, A. V, Shiao, J.-C., Iizuka, Y., & Tzeng, W.-N. (2007). Migratory pattern and habitat use of tropical eels Anguilla spp. (Teleostei: Anguilliformes: Anguillidae) in the Philippines, as revealed by otolith microchemistry. The Raffles Bulletin of Zoology 14(1):141-149.
European Food Safety Authority (EFSA). (2009). Scientific opinion: Animal welfare aspects of husbandry systems for farmed European eel. The European Food Safety Authority Journal, 809(10):1-18.
Fahmi, M. R. (2015). Conservation of genetic tropical fish eel (Anguilla spp) waters in Indonesia. [in English]. Jurnal Penelitian Perikanan Indonesia, 21(1):45-54.
Falah, I. N., Adharini, R. I., & Ratnawati, S. E. (2023). Molecular identification of elvers (Anguilla spp.) from river estuaries in Central Java, Indonesia using DNA barcoding based on mtDNA CO1 sequences. Jurnal Ilmiah Perikanan dan Kelautan, 15(1):121-130.
Fekri, L., Affandi, R., Rahardjo, M. F., Budiardi, T., Simanjuntak C. P. H., Fauzan, T. & Indrayani, I. (2018). The effect of temperature on the physiological condition and growth performance of freshwater eel elver Anguilla bicolor bicolor (McClelland, 1844). Jurnal Akuakultur Indonesia, 17(2):181-190.
Fishbase. (2021, June 12). Anguilla bengalensis and Anguilla marmorata.
Government Regulation of Republic Indonesia No. 82. (2001). Water quality management and control water pollution. Jakarta.
Han, Y-S, Yambot, A. V, Zhang, H, & Hung, C-L. (2012). Sympatric spawning but allopatric distribution of Anguilla japonica and Anguilla marmorata: Temperature- and oceanic current-dependent sieving. Plos One, 7(6):1-11.
Han, C., Dong, S., Li, L., Gao, Q., & Zhou, Y. (2021). DNA barcoding and mini-DNA barcoding reveal mislabeling of salmonids in different distribution channels in the Qingdao area. Journal of Ocean University of China, 20(6):1537-1544.
Hanzen, C., Lucas, M. C., O’Brien, G., Downs, C. T., & Willows-Munro, S. (2020). African freshwater eel species (Anguilla spp.) identification through DNA barcoding. Marine and Freshwater Research, 71(11):1543-1548.
Hebert, P. D. N., Ratnasingham, S., & deWaard, J. R. (2003). Barcoding animal life: Cytochrome C oxidase subunit 1 divergences among closely related species. Proceedings. Biological Sciences / The Royal Society, 270(1):96-99.
Hillis, D. M., & Bull, J. J. (1993). An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology, 42(2):182-192.
Huyen, K. T., & Linh, N. Q. (2020). Phylogenetic analysis of Anguilla marmorata population in Thua Thien Hue, Vietnam based on the cytochrome C oxidase I (COI) gene fragments. AMB Express, 10(1):1-8.
Jellyman, D. J, & Arai T. (2016). Juvenile eels: Upstream migration and habitat use. In: T. Arai (Ed.), Biology and ecology of Anguillid eels. (pp. 143-170) CRC Press Taylor & Francis Group.
Kardin, Sara, L., & Pangerang, U. K. (2016). Some biological aspects of eels (Anguilla sp.) in Mosolo Waters of Wawonii Island, Konawe Archipelago. Manajemen Sumberdaya Perairan, 1(4):355-365.
Kultz, D. (2005). Molecular and evolutionary basis of the cellular stress response. Annual Review Physiology, 2005(67):225-257.
Laudien, J., Flint, N. S., Van Der Bank, F. H., & Brey, T. (2003). Genetic and morphological variation in four populations of the surf clam Donax serra (Röding) from Southern African sandy beaches. Biochemical Systematics and Ecology, 31(7):751-772.
Lawson T. B. (1995). Fundamentals of aquacultural engineering. New York: Chapmanand Hall.
Mallya, Y. J. (2007). The effects of dissolved oxygen on fish growth in aquaculture. UNU-Fisheries Training Programme. 30 pp.
Marnis, H., Syahputra, K., Darmawan, J., Febrianti, D., Tahapari, E., Larashati, S., Iswanto, B., Primanita, E. P. H., Syaifudin, M., & Subangkit, A. T. (2024). DNA barcoding of fish diversity from Batanghari River, Jambi, Indonesia. Fisheries and Aquatic Sciences, 27(2):87-99.
Muchlisin, Z. A., Batubara, A. S., Fadli, N., Muhammadar, A. A., Utami, A. I., Farhana, N., & Siti-Azizah, M. N. (2017). Assessing the species composition of tropical eels (Anguillidae) in Aceh Waters, Indonesia, with DNA barcoding gene cox1. F1000Research, 6(258):1-11.
Mulis. (2015). Enlargement of eel seeds with different types of feed. Scientific Journal of Fisheries and Marine Affairs, 3(1):20-24.
Peninal, S., Subramanian, J., Elavarasi, A., & Kalaiselvam, M. (2017). Genetic identification of marine eels through DNA barcoding from Parangipettai coastal waters. Genomics Data, 11(1):81-84.
Putra, G. I. N., Indrawan, G. S., & Faiqoh, E. (2024). DNA barcoding of cardinalfish (Apogonidae) in Gilimanuk Bay, Bali, Indonesia. Jurnal Ilmiah Perikanan dan Kelautan, 16(1):1-10.
Rasmussen, R. S., Morrissey, M. T., & Hebert, P. D. N. (2009). DNA barcoding of commercially important salmon and trout species (Oncorhynchus and Salmo) from North America. Journal of Agricultural and Food Chemistry, 57(18):8379-8385.
Rasmussen, M.D. & Kellis, M. (2011). Accurate gene-tree reconstruction by learning gene-and species-specific substitution rates across multiple 59 complete genomes. Genome Research, 17(12):1932-1942.
Saanin, H. (1984). Taxonomy and key identification of fish. Volumes 1 and 2. Jakarta: Bina Cipta.
Sadler, K. (1981). The toxicity of ammonia to the European eel (Anguilla anguilla L.). Aquaculture, 26(1-2):173-181.
Silfvergrip, A. M. (2009). Report 5943 - CITES identification guide to the freshwater eels (Anguillidae) with focus on the European eel Anguilla anguilla.
Stecher, G., Knyaz, C., Tamura, K., & Kumar, S. (2020). Molecular evolutionary genetics analysis (MEGA) for macOS. Molecular Biology and Evolution, 37(4):1237-1239.
Syaifudin, M., Bekaert, M., Taggart, J., Hulata, G., D’Cotta, H., Baroiller, J., Penman, D., & McAndrew, B. (2017a). DNA typing across ten tilapia species using cytochrome C oxidase subunit I (COI) in Abstracts: Twelfth International Symposium on Genetics in Aquaculture 2015.. Aquaculture, 472(4/1):125-125.
Syaifudin, M., Gultom, E. T., & Wijayanti, M. (2023). DNA authentication of Indonesian leaffish Pristolepis grooti from Kelekar River and Ogan River in South Sumatra based on cytochrome C oxidase subunit I (COI) Gene. Journal of Tropical Biodiversity and Biotechnology, 8(2):1-11.
Syaifudin, M., Jubaedah, D., Muslim, M., & Daryani, A. (2017b). DNA authentication of Asian redtail catfish Hemibagrus nemurus from Musi and Penukal River, South Sumatra Indonesia. Genetics of Aquatic Organisms, 1(12):43-48.
Syaifudin, M., Jubaedah, D., Taqwa, F. H., & Octaviani, R. (2021). Phylogenetic of marble goby (Oxyeleotris marmorata Blkr.) in South Sumatra based on cytochrome C oxidase subunit I (COI) gene. Genetics of Aquatic Organisms, 6(1):1-6.
Tamura, K., Stecher, G., & Kumar, S. (2021). MEGA1: Molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution, 38(7):3022-3027.
Ward, R. D., Zemlak, T. S, Innes, B. H, Last, P. R., Hebert, P. D. N. (2005). DNA barcoding Australia’s fish species. Philosophical Transactions of the Royal Society B, 360(1462):1847–57.
Watanabe, S., Aoyama, J., Miller, M. J., Ishikawa, S., Feunteun, E., & Tsukamoto, K. (2008). Evidence of population structure in the giant mottled eel, Anguilla marmorata, using total number of vertebrae. Copeia, 2008(3):680-688.
Wibowo, A., Hubert, N., Dahruddin, H., Steinke, D., Suhaimi, R. A., Samuel, Atminarso, D., Anggraeni, D. P., Trismawanti, I., Baumgartner, L. J., & Ning, N. (2021). Assessing temporal patterns and species composition of glass eel (Anguilla spp.) cohorts in Sumatra and Java using DNA barcodes. Diversity, 13(5):1-15.
Zan, N. D., Sarbini, A., Taha, H., Tan, I. V., Azri, A., Kahar, R., Metali, F., Ahmad, N., & Arai, T. (2020). Occurrence and ecological implication of a tropical anguillid eel, Anguilla marmorata, in Brunei Darussalam, Borneo Island. Zoologia, 37(6):1-7.