EXPLORATION OF INDIGENOUS BACTERIA WITH POTENTIAL FOR TOTAL AMMONIA NITROGEN DEGRADATION FROM RUBBER WASTEWATER AND PHYLOGENETIC TREE CONSTRUCTION BASED ON 16S rRNA GENE SEQUENCES
DOI:
https://doi.org/10.26418/jpmipa.v16i3.93436Keywords:
Bacteria, Bioremediation, Degradation, Rubber Wastewater, Total Amonia Nitrogen (TAN)Abstract
Rubber wastewater contains total ammonia nitrogen (TAN) at levels that can potentially cause pollution if discharged into water bodies without proper treatment. TAN content is one of the commonly used parameters for assessing water quality. Certain groups of bacteria are capable of naturally degrading TAN in the environment. This study aimed to isolate bacteria from rubber wastewater, evaluate their potential to degrade TAN, and identify the most effective isolate at the molecular level. Samples were collected from wastewater treatment ponds using random purposive sampling based on specific criteria. A survey method was used to determine sampling locations, while experimental methods were applied in the laboratory. Bacterial isolation was conducted using the serial dilution technique, followed by inoculation with the pour plate technique. Pure isolates were obtained using the streak plate technique. The degradation potential of the bacterial isolates was tested by inoculating 10% v/v of each isolate into 200 mL of rubber wastewater, followed by incubation at room temperature for six days. TAN levels were analyzed using the phenate method with a spectrophotometer, following the Indonesian National Standard SNI 06-6989.30-2005. Three bacterial isolates (NS-1, NS-2, and NB-1) were obtained from the rubber wastewater. All three isolates demonstrated potential in reducing TAN levels, with final TAN concentrations after incubation ranging from 6.75 to 4.8 mg/L, corresponding to a reduction of 52.50"“66.90%. The most effective isolate in reducing TAN was NS-2. Molecular identification using the PCR-seq method of the 16S rRNA gene revealed that isolate NS-2 showed 99.86"“99.93% sequence identity with Enterobacter sp. This study provides a basis for the development of bioremediation technologies for rubber wastewater treatment to improve aquatic environmental quality.References
BPS-Sumbar. (2025). Produksi Perkebunan Rakyat Menurut Jenis Tanaman di Provinsi Sumatera Barat (ribu ton), 2024.
BPS. (2024). Indonesian Rubber Statistic. 1–120.
BSN. (2005). Cara uji ammonia dengan spektrofotometer secara fenat. SNI. 06-6989.30-2005. Badan Standarisasi Nasional, 6.
BSN. (2008). Metoda Pengambilan Contoh Air Permukaan, SNI 6989.59:2008. Badan Standardisasi Nasional, 1–19. http://ciptakarya.pu.go.id/plp/upload/peraturan/SNI_-6989-59-2008-_Metoda-Pengambilan-Contoh-Air-Limbah.pdf
Chhasatiya, P., & Patel, M. (2018). Removal of ammonical nitrogen from industrial effluent. International Journal of Advance Research, 4(3), 1108–1110. www.IJARIIT.com
Detho, A. … Din, M. F. M. (2022). Ammoniacal nitrogen, chemical oxygen demand, and color reduction in rubber processing industry effluent using zeolite. Desalination and Water Treatment, 270, 185–193. https://doi.org/10.5004/dwt.2022.28761
Duan, S. … Zheng, S. (2022). Heterotrophic nitrifying bacteria in wastewater biological nitrogen removal systems: A review. CREST, 52(13), 2302–2338. https://doi.org/10.1080/10643389.2021.1877976
Et, S. (2010). Ammonium Concentrations in Produced Waters from a Mesothermic Oil Field Subjected to Nitrate Injection Decrease through Formation of Denitrifying Biomass and Anammox Activity. Applied and Environmental Microbiology, 76(15), 4977–4987. https://doi.org/10.1128/AEM.00596-10
Febria, F. A. … Febrion, C. (2024). Degradation of low-density polyethylene by a novel strain of bacteria isolated from the plastisphere of marine ecosystems. Global Journal of Environmental Science and Management, 10(2), 805–820. https://doi.org/10.22034/GJESM.2024.02.24
Ge, S. … Peng, Y. (2015). Detection of nitrifiers and evaluation of partial nitrification for wastewater treatment: A review. Chemosphere, 140, 85–98. https://doi.org/10.1016/j.chemosphere.2015.02.004
Komarudin, N. A. … Velasco, A. B. (2022). Exposure to Ammonia Concentration from the Processing of Crumb Rubber on Environmental Quality: A Review. Asia Pacific Journal of Sustainable Agriculture, Food and Energy, 10(1), 22–27. https://doi.org/10.36782/APJSAFE.V10I1.127
Li, S. … Yu, J. (2024). Nitrogen removal by heterotrophic nitrification-aerobic denitrification bacteria: A review. Desalination and Water Treatment, 317(February), 100227. https://doi.org/10.1016/j.dwt.2024.100227
Lukkari, I. … Christian, G. D. (1993). Determination of total ammonium-nitrogen and free ammonia in a fermentation medium by sequential injection analysis. Fresenius’ Journal of Analytical Chemistry, 346(6–9), 813–818. https://doi.org/10.1007/BF00321296
Luo, L. … Xu, Q. (2016). Isolation, identification, and optimization of culture conditions of a bioflocculant-producing bacterium bacillus megaterium sp1 and its application in aquaculture wastewater treatment. BioMed Research International, 2016. https://doi.org/10.1155/2016/2758168
Nurisman, E. … Rosmania. (2023). Reduksi Kadar Amoniak Limbah Cair Industri Karet pada Airlift Bioreaktor dengan Bakteri Brevundimonas diminuta. JST (Jurnal Sains Dan Teknologi), 12(2), 376–384. https://doi.org/10.23887/jstundiksha.v12i2.48396
Odokuma, L. O., & Akponah, E. (2008). Response of Nitrosomonas, Nitrobacter and Escherichia coli to drilling fluids. Journal of Cell and Animal Biology, 2(2), 43–054. http://www.academicjournals.org/JCAB
Ouyang, L. … Li, S. (2024). Insights into the Nitrogen Removal Mechanism of Heterotrophic Nitrification and Aerobic Denitrification Bacterium Delfitia sp. B7. Water (Switzerland), 16(21), 3042. https://doi.org/10.3390/W16213042/S1
Paśmionka, I. B. … de Prá, M. C. (2021). Toxic Effect of Ammonium Nitrogen on the Nitrification Process and Acclimatisation of Nitrifying Bacteria to High Concentrations of NH4-N in Wastewater. Energies , 14(17). https://doi.org/10.3390/EN14175329
Peng, Y., & Zhu, G. (2006). Biological nitrogen removal with nitrification and denitrification via nitrite pathway. Applied Microbiology and Biotechnology, 73(1), 15–26. https://doi.org/10.1007/s00253-006-0534-z
Pisani, O. … J. Cole, K. (2024). USDA LTAR Common Experiment measurement: Dissolved ammonia (NH3) concentration v1. Protocols.Io. https://doi.org/10.17504/PROTOCOLS.IO.J8NLK8B61L5R/V1
Rana, A. … Ramakrishnan, B. (2019). Enzymology of the nitrogen cycle and bioremediation of toxic nitrogenous compounds. Smart Bioremediation Technologies: Microbial Enzymes, October 2020, 45–61. https://doi.org/10.1016/B978-0-12-818307-6.00003-2
Wimalaweera, I. P. … Weragoda, S. (2024). Enhanced Pretreatment of Natural Rubber Industrial Wastewater Using Magnetic Seed Coagulation with Ca(OH)2. Water (Switzerland), 16(6), 847. https://doi.org/10.3390/W16060847/S1
Ye, G. zi … Ouyang, P. kai. (2010). Isolation of NH₄+-tolerant mutants of Actinobacillus succinogenes for succinic acid production by continuous selection. Journal of Microbiology and Biotechnology, 20(8), 1219–1225. https://doi.org/10.4014/JMB.1003.03030
Zhang … Shi, K. (2022). Indole-Acetic Acid Promotes Ammonia Removal Through Heterotrophic Nitrification, Aerobic Denitrification With Mixed Enterobacter sp. Z1 and Klebsiella sp. Z2. Frontiers in Microbiology, 13(July). https://doi.org/10.3389/fmicb.2022.929036
Zhang, X. … Wang, X. (2015). Chlorine/UV induced photochemical degradation of total ammonia nitrogen (TAN) and process optimization. RSC Advances, 5(78), 63793–63799. https://doi.org/10.1039/C5RA10654K
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Jurnal Pendidikan Matematika dan IPA

This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution 4.0 International License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).

This work is licensed under a Creative Commons Attribution 4.0 International License