DNA Metabarcoding Reveals Marine Microbiota Composition as Indicators of Anthropogenic Impact in Karimunjawa National Park
DOI:
https://doi.org/10.24002/biota.v11i2.13185Keywords:
16S rDNA, Anthropogenic, Diversity, DNA Metabarcoding, MicroorganismsAbstract
The Karimunjawa Islands in Central Java, Indonesia, are a designated marine national park experiencing increasing anthropogenic pressures. To address these concerns, this study investigates the diversity of marine microbiota in the coastal waters of the Karimunjawa Islands, utilizing DNA metabarcoding techniques to assess their potential as indicators of environmental change. Environmental DNA (eDNA) samples were collected from both high and low human-impacted site. Samples were analysed using Next Generation Sequencing (NGS) technology targeting the 16S gene. Analysis of these samples revealed a high diversity of marine microbiota, comprising 11 phyla, 11 classes, 11 orders, and 12 families. In locations subjected to anthropogenic pressures, the microbial community was dominated by potentially pathogenic taxa, particularly those from the families Arcobacteraceae and Nitrincolaceae, which are commonly associated with degraded water quality and adverse conditions for aquatic life. Conversely, low human-impacted sites exhibited a predominance of Cyanobiaceae, a group known for its ecological roles that support a healthy aquatic environment. The study's findings underscore the importance of marine microbiota as a valuable bioindicator for monitoring environmental health in coastal areas. These results highlight the need for ongoing monitoring and conservation efforts to preserve the delicate balance of the marine ecosystems in the Karimunjawa Islands.
References
Acharya-Patel, N., Allison, M. J., & Helbing, C. C. (2022). Environmental DNA: Revolutionizing ecological assessments with genomics. In Genomics and the Global Bioeconomy. https://doi.org/10.1016/B978-0-323-91601-1.00004-3
Adeyemo, S. M., & Onilude, A. A. (2014). Molecular Identification of Lactobacillus plantarum Isolated from Fermenting Cereals. International Journal for Biotechnology and Molecular Biology Research, 5(6).
Ahmed, M. A., & Campbell, B. J. (2025). Genome-resolved adaptation strategies of Rhodobacterales to changing conditions in the Chesapeake and Delaware Bays. Applied and Environmental Microbiology, 91(2), e02357-24. https://doi.org/10.1128/aem.02357-24
Alwi, M., Mutaqin, B. W., & Marfai, M. A. (2023). Shoreline Dynamics in the Very Small Islands of Karimunjawa – Indonesia: a Preliminary Study. Geoplanning, 10(1). https://doi.org/10.14710/geoplanning.10.1.73-82
Amin, M., Pramujisunu, Y., Cahyani, N. K. D., Mukti, A. T., Lamid, M., Ali, M., & Eroldoğan, O. T. (2023). The structure, composition, and predicted microbiome functional genes in Pacific white shrimp (Litopenaeus vannamei) grow-out ponds with different survival rates through high-throughput sequencing. Aquatic Sciences, 85(3). https://doi.org/10.1007/s00027-023-00979-3
Barnes, M. A., & Turner, C. R. (2016). The ecology of environmental DNA and implications for conservation genetics. In Conservation Genetics (Vol. 17, Issue 1). https://doi.org/10.1007/s10592-015-0775-4
Bischoff, V., Zucker, F., & Moraru, C. (2020). Marine Bacteriophages. In Encyclopedia of Virology: Volume 1-5, Fourth Edition (Vols. 1–5). https://doi.org/10.1016/B978-0-12-809633-8.20988-6
Cahyani, N. K. D., Anggoro, A. W., Al Malik, M. D., Subhan, B., Sani, L. M. I., & Madduppa, H. (2024). Inventorizing marine biodiversity using eDNA data from Indonesian coral reefs: comparative high throughput analysis using different bioinformatic pipelines. Marine Biodiversity, 54(3). https://doi.org/10.1007/s12526-024-01432-w
Callahan, B. J., McMurdie, P. J., Rosen, M. J., Han, A. W., Johnson, A. J. A., & Holmes, S. P. (2016). DADA2: High-resolution sample inference from Illumina amplicon data. Nature Methods, 13(7). https://doi.org/10.1038/nmeth.3869
Caporaso, J. G., Lauber, C. L., Walters, W. A., Berg-Lyons, D., Lozupone, C. A., Turnbaugh, P. J., Fierer, N., & Knight, R. (2011). Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proceedings of the National Academy of Sciences of the United States of America, 108(SUPPL. 1). https://doi.org/10.1073/pnas.1000080107
Chang, H., Ye, T., Xie, Z., & Liu, X. (2025). Application of Environmental DNA in Aquatic Ecosystem Monitoring: Opportunities, Challenges and Prospects. Water, 17(5). https://doi.org/10.3390/w17050661
Collado, L., Inza, I., Guarro, J., & Figueras, M. J. (2008). Presence of Arcobacter spp. in environmental waters correlates with high levels of fecal pollution. Environmental Microbiology, 10(6). https://doi.org/10.1111/j.1462-2920.2007.01555.x
Collado, L., & Figueras, M. J. (2011). Taxonomy, epidemiology, and clinical relevance of the genus Arcobacter. Clinical Microbiology Reviews, 24(1), 174–192. https://doi.org/10.1128/CMR.00034-10
Curson, A. R. J., Williams, B. T., Pinchbeck, B. J., Sims, L. P., Martínez, A. B., Rivera, P. P. L., Kumaresan, D., Mercadé, E., Todd, J. D., & Johnston, A. W. B. (2017). DSYB catalyses the key step of dimethylsulfoniopropionate biosynthesis in many phytoplankton. Nature Microbiology, 2, 17009. https://doi.org/10.1038/nmicrobiol.2017.9
Dong, Y., Gao, J., Wu, Q., Ai, Y., Huang, Y., Wei, W., Sun, S., & Weng, Q. (2020). Co-occurrence pattern and function prediction of bacterial community in Karst cave. BMC Microbiology, 20(1). https://doi.org/10.1186/s12866-020-01806-7
Dupont, C. L., Rusch, D. B., Yooseph, S., Lombardo, M.-J., Alexander Richter, R., Valas, R., Novotny, M., Yee-Greenbaum, J., Selengut, J. D., Haft, D. H., Halpern, A. L., Lasken, R. S., Nealson, K., Friedman, R., & Craig Venter, J. (2012). Genomic insights to SAR86, an abundant and uncultivated marine bacterial lineage. The ISME Journal, 6(6), 1186–1199. https://doi.org/10.1038/ismej.2011.189
Fahma, T. L. N., Cahyani, N. K. D., Jumari, J., Hariyati, R., Soeprobowati, T. R. (2024). Environmental DNA Approach to Identify Protists Community in Sediment of Balekambang Lake, Indonesia, Using 18S rRNA Gene. In: Haddout, S., Priya, K., Hoguane, A.M. (eds) Proceedings of The 2nd International Conference on Climate Change and Ocean Renewable Energy. CCORE 2022. Springer Proceedings in Earth and Environmental Sciences. Springer, Cham. https://doi.org/10.1007/978-3-031-71555-6_25
Fifer, J. E., Bui, V., Berg, J. T., Kriefall, N., Klepac, C., Bentlage, B., & Davies, S. W. (2022). Microbiome Structuring Within a Coral Colony and Along a Sedimentation Gradient. Frontiers in Marine Science, 8. https://doi.org/10.3389/fmars.2021.805202
Fleisher, J. M., Fleming, L. E., Solo-Gabriele, H. M., Kish, J. K., Sinigalliano, C. D., Plano, L., Elmir, S. M., Wang, J. D., Withum, K., Shibata, T., Gidley, M. L., Abdelzaher, A., He, G., Ortega, C., Zhu, X., Wright, M., Hollenbeck, J., & Backer, L. C. (2010). The BEACHES study: Health effects and exposures from non-point source microbial contaminants in subtropical recreational marine waters. International Journal of Epidemiology, 39(5). https://doi.org/10.1093/ije/dyq084
Flombaum, P., Gallegos, J. L., Gordillo, R. A., Rincón, J., Zabala, L. L., Jiao, N., Karl, D. M., Li, W. K. W., Lomas, M. W., Veneziano, D., Vera, C. S., Vrugt, J. A., & Martiny, A. C. (2013). Present and future global distributions of the marine Cyanobacteria Prochlorococcus and Synechococcus. Proceedings of the National Academy of Sciences, 110(24), 9824–9829. https://doi.org/10.1073/pnas.1307701110
Gaines, W. L., Harrod, R. J., & Lehmkuhl, J. F. (1999). Monitoring biodiversity: Quantification and interpretation. General Technical Reports of the US Department of Agriculture, Forest Service, PNW-GTR-443.
Giovannoni, S. J., & Stingl, U. (2005). Molecular diversity and ecology of microbial plankton. Nature, 437(7057), 343–348. https://doi.org/10.1038/nature04158
Gold, Z., Sprague, J., Kushner, D. J., Marin, E. Z., & Barber, P. H. (2021). eDNA metabarcoding as a biomonitoring tool for marine protected areas. PLoS ONE, 16(2 February 2021). https://doi.org/10.1371/journal.pone.0238557
Halim M. A. R., Soeprobowati T. R., Hadiyanto H., (2022). Identification of potential water pollution in coastal areas from anthropogenic activities in Karimunjawa National Park. AACL Bioflux, 15(6), 2969-2981.
Hernández-Zulueta, J., Rubio-Bueno, S., Zamora-Tavares, M. del P., Vargas-Ponce, O., Rodríguez-Troncoso, A. P., & Rodríguez-Zaragoza, F. A. (2024). Metabarcoding the Bacterial Assemblages Associated with Toxopneustes roseus in the Mexican Central Pacific. Microorganisms, 12(6). https://doi.org/10.3390/microorganisms12061195
Huang, J., Xin, Y., Cao, X., & Zhang, W. (2011). Phylogenetic diversity and characterization of 2-haloacid degrading bacteria from the marine sponge Hymeniacidon perlevis. World Journal of Microbiology and Biotechnology, 27(8). https://doi.org/10.1007/s11274-010-0636-8
Iber, B. T., & Kasan, N. A. (2021). Recent advances in Shrimp aquaculture wastewater management. In Heliyon (Vol. 7, Issue 11). https://doi.org/10.1016/j.heliyon.2021.e08283
Isokpehi, R. D., Kim, Y., Krejci, S. E., & Trivedi, V. D. (2024). Ecological Trait-Based Digital Categorization of Microbial Genomes for Denitrification Potential. Microorganisms, 12(4). https://doi.org/10.3390/microorganisms12040791
Kadarusman, K., Rachmawati, R., Setyawidati, N. A. R., Sektiana, S. P., Tapilatu, R. F., Albasri, H., Nurdin, E., Saputra, R. S. H., Noviendri, D., & Nursid, M. (2019). Sumber Daya Hayari Maritim. Amafrad Press, Jakarta.
Kandlikar, G. S., Gold, Z. J., Cowen, M. C., Meyer, R. S., Freise, A. C., Kraft, N. J. B., Moberg-Parker, J., Sprague, J., Kushner, D. J., & Curd, E. E. (2018). Ranacapa: An R package and shiny web app to explore environmental DNA data with exploratory statistics and interactive visualizations . F1000Research, 7. https://doi.org/10.12688/f1000research.16680.1
Kersters, K., De Vos, P., Gillis, M., Swings, J., Vandamme, P., & Stackebrandt, E. (2006). Introduction to the Proteobacteria. In The Prokaryotes. https://doi.org/10.1007/0-387-30745-1_1
Khan, S., & Malik, A. (2021). Exploring the Diversity of Marine Microbiome in Response to Changes in the Environment. In Microbiomes and the Global Climate Change. https://doi.org/10.1007/978-981-33-4508-9_6
Liu, C., Yan, M., Xiong, W., Li, N., & Gao, L. (2024). Editorial: Applications of environmental DNA in the aquatic ecosystem management of East Asia. Frontiers in Marine Science, Volume 11-2024. https://doi.org/10.3389/fmars.2024.1473463
Marchesi, J. R., & Ravel, J. (2015). The vocabulary of microbiome research: a proposal. Microbiome, 3(1). https://doi.org/10.1186/s40168-015-0094-5
Marwayana, O. N., Gold, Z., Meyer, C. P., & Barber, P. H. (2022). Environmental DNA in a global biodiversity hotspot: Lessons from coral reef fish diversity across the Indonesian archipelago. Environmental DNA, 4(1). https://doi.org/10.1002/edn3.257
McIlroy, S. J., & Nielsen, P. H. (2014). The family saprospiraceae. In The Prokaryotes: Other Major Lineages of Bacteria and The Archaea (Vol. 9783642389542). https://doi.org/10.1007/978-3-642-38954-2_138
McMurdie, P. J., & Holmes, S. (2013). Phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data. PLoS ONE, 8(4). https://doi.org/10.1371/journal.pone.0061217
Michán, C., Blasco, J., & Alhama, J. (2021). High-throughput molecular analyses of microbiomes as a tool to monitor the wellbeing of aquatic environments. In Microbial Biotechnology (Vol. 14, Issue 3). https://doi.org/10.1111/1751-7915.13763
Moran, M. A., Durham, B. P., Kubler, J. E., et al. (2022). Sulfur metabolites in the pelagic ocean. Nature Reviews Microbiology, 20, 482–496. https://doi.org/10.1038/s41579-022-00715-0
Morgan, M. C., Boyette, M., Goforth, C., Sperry, K. V., & Greene, S. R. (2009). Comparison of the Biolog OmniLog Identification System and 16S ribosomal RNA gene sequencing for accuracy in identification of atypical bacteria of clinical origin. Journal of Microbiological Methods, 79(3). https://doi.org/10.1016/j.mimet.2009.10.005
Namirimu, T., Kim, Y. J., Park, M.-J., Lim, D., Lee, J.-H., & Kwon, K. K. (2022). Microbial Community Structure and Functional Potential of Deep-Sea Sediments on Low Activity Hydrothermal Area in the Central Indian Ridge. Frontiers in Marine Science, Volume 9-2022. https://doi.org/10.3389/fmars.2022.784807
Oh, J. W., Pushparaj, S. S. C., Muthu, M., & Gopal, J. (2023). Review of Harmful Algal Blooms (HABs) Causing Marine Fish Kills: Toxicity and Mitigation. In Plants (Vol. 12, Issue 23). https://doi.org/10.3390/plants12233936
Orel, N., Fadeev, E., Klun, K., Ličer, M., Tinta, T., & Turk, V. (2022). Bacterial indicators are ubiquitous members of pelagic microbiome in anthropogenically impacted coastal ecosystem. Frontiers in Microbiology, 12, 765091. https://doi.org/10.3389/fmicb.2021.765091
O’Hara, C. C., Frazier, M., & Halpern, B. S. (2021). At-risk marine biodiversity faces extensive, expanding, and intensifying human impacts. Science, 372(6537). https://doi.org/10.1126/science.abe6731
Paerl, H. W., & Otten, T. G. (2013). Harmful Cyanobacterial Blooms: Causes, Consequences, and Controls. Microbial Ecology, 65(4). https://doi.org/10.1007/s00248-012-0159-y
PÁez-Osuna, F. (2001). The environmental impact of shrimp aquaculture: Causes, effects, and mitigating alternatives. In Environmental Management (Vol. 28, Issue 1). https://doi.org/10.1007/s002670010212
Pérez-Cataluña, A., Salas-Massó, N., Diéguez, A. L., Balboa, S., Lema, A., Romalde, J. L., & Figueras, M. J. (2018). Revisiting the taxonomy of the genus Arcobacter: Getting order from the chaos. Frontiers in Microbiology, 9, 2077. https://doi.org/10.3389/fmicb.2018.02077
Pohlner, M., Dlugosch, L., Wemheuer, B., Mills, H., Engelen, B., & Reese, B. K. (2019). The majority of active Rhodobacteraceae in marine sediments belong to uncultured genera: A molecular approach to link their distribution to environmental conditions. Frontiers in Microbiology, 10(APR). https://doi.org/10.3389/fmicb.2019.00659
Pujalte, M. J., Lucena, T., Ruvira, M. A., Arahal, D. R., & Macián, M. C. (2014). The family Rhodobacteraceae. In The Prokaryotes: Alphaproteobacteria and Betaproteobacteria (Vol. 9783642301971). https://doi.org/10.1007/978-3-642-30197-1_377
Qu, L., Li, M., Gong, F., He, L., Li, M., Zhang, C., Yin, K., & Xie, W. (2024). Oxygen-driven divergence of marine group II archaea reflected by transitions of superoxide dismutases. Microbiology Spectrum, 12(1). https://doi.org/10.1128/spectrum.02033-23
Rathore, S. S., Chandravanshi, P., Chandravanshi, A., & Jaiswal, K. (2016). Eutrophication: Impacts of Excess Nutrient Inputs on Aquatic Ecosystem. IOSR Journal of Agriculture and Veterinary Science, 09(10). https://doi.org/10.9790/2380-0910018996
Rolph, H. J., Lennon, A., Riggio, M. P., Saunders, W. P., MacKenzie, D., Coldero, L., & Bagg, J. (2001). Molecular identification of microorganisms from endodontic infections. Journal of Clinical Microbiology, 39(9). https://doi.org/10.1128/JCM.39.9.3282-3289.2001
Ruppert, K. M., Kline, R. J., & Rahman, M. S. (2019). Past, present, and future perspectives of environmental DNA (eDNA) metabarcoding: A systematic review in methods, monitoring, and applications of global eDNA. In Global Ecology and Conservation (Vol. 17). https://doi.org/10.1016/j.gecco.2019.e00547
Satomi, M., & Fujii, T. (2014). The family oceanospirillaceae. In The Prokaryotes: Gammaproteobacteria (Vol. 9783642389221). https://doi.org/10.1007/978-3-642-38922-1_286
Scanlan, D. J., Ostrowski, M., Mazard, S., et al. (2009). Ecological genomics of marine picocyanobacteria. Microbiology and Molecular Biology Reviews, 73(2), 249–299. https://doi.org/10.1128/MMBR.00035-08
Shan, H., Du, Y., Li, T., Wang, F., Li, H., & Wang, H. (2024). Bacterial Community Characteristics and Roles in Nitrogen Transformation in Industrial Farming Systems of Litopenaeus vannamei. Journal of Marine Science and Engineering, 12(5). https://doi.org/10.3390/jmse12050787
Siagian, R. A. S., Sabdono, A., Sunaryo, S., Trianto, A., & Dirgantara, D. (2022). Nutrient Enrichment Impact of Wastewater Shrimp Ponds on Coral Reefs of Nyamplungan Village, Karimunjawa. Ilmu Kelautan: Indonesian Journal of Marine Sciences, 27(3). https://doi.org/10.14710/ik.ijms.27.3.267-278
Singh, J. S., Kumar, A., Rai, A. N., & Singh, D. P. (2016). Cyanobacteria: A precious bio-resource in agriculture, ecosystem, and environmental sustainability. In Frontiers in Microbiology (Vol. 7, Issue APR). https://doi.org/10.3389/fmicb.2016.00529
Stewart, J. R., Gast, R. J., Fujioka, R. S., Solo-Gabriele, H. M., Meschke, J. S., Amaral-Zettler, L. A., Del Castillo, E., Polz, M. F., Collier, T. K., Strom, M. S., Sinigalliano, C. D., Moeller, P. D. R., & Holland, A. F. (2008). The coastal environment and human health: Microbial indicators, pathogens, sentinels and reservoirs. Environmental Health: A Global Access Science Source, 7(SUPPL. 2). https://doi.org/10.1186/1476-069X-7-S2-S3
Terzin, M., Laffy, P. W., Robbins, S., Yeoh, Y. K., Frade, P. R., Glasl, B., Webster, N. S., & Bourne, D. G. (2024). The road forward to incorporate seawater microbes in predictive reef monitoring. In Environmental Microbiome (Vol. 19, Issue 1). https://doi.org/10.1186/s40793-023-00543-4
Uljanovas, D., Gölz, G., Fleischmann, S., Kudirkiene, E., Kasetiene, N., Grineviciene, A., Tamuleviciene, E., Aksomaitiene, J., Alter, T., & Malakauskas, M. (2023). Genomic Characterization of Arcobacter butzleri Strains Isolated from Various Sources in Lithuania. Microorganisms, 11(6). https://doi.org/10.3390/microorganisms11061425
Venâncio, I., Luís, Â., Domingues, F., Oleastro, M., Pereira, L., & Ferreira, S. (2022). The Prevalence of Arcobacteraceae in Aquatic Environments: A Systematic Review and Meta-Analysis. In Pathogens (Vol. 11, Issue 2). https://doi.org/10.3390/pathogens11020244
Wickham, Hadley. (2009). Ggplot2 : elegant graphics for data analysis. Journal of Statistical Software, 35(July).
Wilkins, L. G. E., Leray, M., O’Dea, A., Yuen, B., Peixoto, R. S., Pereira, T. J., Bik, H. M., Coil, D. A., Duffy, J. E., Herre, E. A., Lessios, H. A., Lucey, N. M., Mejia, L. C., Rasher, D. B., Sharp, K. H., Sogin, E. M., Thacker, R. W., Thurber, R. V., Wcislo, W. T., … Eisen, J. A. (2019). Host-associated microbiomes drive structure and function of marine ecosystems. PLoS Biology, 17(11). https://doi.org/10.1371/journal.pbio.3000533
Williams, B. T., Cowles, K., Bermejo Martínez, A., Curson, A. R. J., Zheng, Y., Liu, J., Newton-Payne, S., Hind, A. J., Li, C.-Y., Rivera, P. P. L., Carrión, O., Liu, J., Spurgin, L. G., Brearley, C. A., Mackenzie, B. W., Pinchbeck, B. J., Peng, M., Pratscher, J., Zhang, X.-H., … Todd, J. D. (2019). Bacteria are important dimethylsulfoniopropionate producers in coastal sediments. Nature Microbiology, 4(11), 1815–1825. https://doi.org/10.1038/s41564-019-0527-1
Zada, S., Zhou, H., Xie, J., Hu, Z., Ali, S., Sajjad, W., & Wang, H. (2021). Bacterial degradation of pyrene: Biochemical reactions and mechanisms. In International Biodeterioration and Biodegradation (Vol. 162). https://doi.org/10.1016/j.ibiod.2021.105233
Zahra, Z., Choo, D. H., Lee, H., & Parveen, A. (2020). Cyanobacteria: Review of current potentials and applications. In Environments - MDPI (Vol. 7, Issue 2). https://doi.org/10.3390/environments7020013
Zhang, Y., Zhao, Z., Dai, M., Jiao, N., & Herndl, G. J. (2020). Drivers shaping the diversity and biogeography of total and active bacterial communities in the South China Sea. Molecular Ecology, 29(7), 1212–1228.
Zhu, W., Liu, J., Li, Q., Gu, P., Gu, X., Wu, L., Gao, Y., Shan, J., Zheng, Z., & Zhang, W. (2022). Effects of Nutrient Levels on Microbial Diversity in Sediments of a Eutrophic Shallow Lake. Frontiers in Ecology and Evolution, 10. https://doi.org/10.3389/fevo.2022.909983
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