Diesel effect on the growth and biochemical composition of Chlorella spp.: mixotrophy and microalgae-bacteria consortium
DOI:
https://doi.org/10.5281/zenodo.18487497Keywords:
diesel, hydrocarbon removal, microalgae, organic macromolecules, toxic effectAbstract
Environmental pollution by organic compounds is constantly worsening with the increase in mining and industrial activities, which requires the development of treatment technologies to minimize the anthropogenic impact on ecosystems. Therefore, in this study, the effect of diesel on the growth and biochemical composition of Chlorella spp. was evaluated using batch tests at laboratory scale with treatments of 0.25 (T1), 0.50 (T2) and 1.00 (T3) % v/v of diesel. Chlorophyll a concentrations were between 0.51 and 8.52 µg/mL, chlorophyll b between 0.15 and 3.09 µg/mL, total chlorophyll between 0.66 and 11.32 µg/mL and carotenoids between 0.24 and 3.66 µg/mL; while total protein concentrations ranged from 160.32 to 818.44 µg/mL, total carbohydrates from 43.80 to 209.22 µg/mL, and total lipids from 110.30 to 715.31 µg/mL. Changes in the photosynthetic pigment contents of the microalgae (T1 and T2) were observed, as well as stimulation of growth (T1) and production of biomolecules (all treatments), probably due to the mixotrophic growth of the microalgae and/or the symbiotic action with the associated bacteria (microbial consortium). The best performance in the degradation of total petroleum hydrocarbons was obtained at the lowest proportion of diesel applied (T1: 75.8 %). The use of microalgae in the form of mixotrophic culture or in microbial consortia is highly versatile for hydrocarbon degradation, reducing pollution and enabling the production of commercially important metabolites.
Key words: diesel; hydrocarbon removal; microalgae; organic macromolecules; toxic effect.
References
Referencias
Aldaby, E. S. E. y Mawad, A. M. M. (2019). Pyrene biodegradation capability of two different microalgal strains. Global Nest Journal, 21(3), 290-295. https://doi.org/10.30955/gnj.002767
Al-Wasify, R. S. y Hamed, S. R. (2014). Bacterial biodegradation of crude oil using local isolates. International Journal of Bacteriology, 2014, 863272. https://doi.org/10.1155/2014/863272
Ambaye, T. G., Chebbi, A., Formicola, F., Prasad, S., Gomez, F. H., Franzetti, A. y Vaccari, M. (2022). Remediation of soil polluted with petroleum hydrocarbons and its reuse for agriculture: recent progress, challenges, and perspectives. Chemosphere, 293, 133572. https://doi.org/10.1016/j.chemosphere.2022.133572
American Public Health Association (APHA), American Water Works Association (AWWA) y Water Environment Federation (WEF). (2023). Standard methods for the examination of water and wastewater. (24th edition). W. C. Lipps, E. B. Braun-Howland y T. E. Baxter (Eds.). American Public Health Association.
Bento, F. M., de Oliveira-Camargo, F. A., Okeke, B. y Frankenberger-Júnior, W. T. (2003). Bioremediation of soil contaminated by diesel oil. Brazilian Journal of Microbiology, 34(1), 65-68. http://doi.org/10.1590/S1517-83822003000500022
Bligh, E. G. y Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology, 37(8), 911-917. https://doi.org/10.1139/o59-099
Chaudhuri, U. R. (2011). Fundamentals of petroleum and petrochemical engineering. CRC Press.
Chekroun, K. B., Sánchez, E. y Baghour, M. (2014). The role of algae in bioremediation of organic pollutants. International Research Journal of Public and Environmental Health, 1(2), 19-32. https://journalissues.org/wp-content/uploads/2014/07/Chekourn-et-al.pdf
Cortez-Mago, R., Guevara, M., Vásquez, A. y Lodeiros-Seijo, C. (2007). Influencia del petróleo crudo en el crecimiento de microalgas del nororiente de Venezuela. Boletín del Centro de Investigaciones Biológicas, 41(4), 471-483. https://produccioncientificaluz.org/index.php/boletin/article/view/100
Das, P., AbdulQuadir, M., Thaher, M., Khan, S., Chaudhary, A. K., Alghasal, G. y Al-Jabri, H. M. (2019). Microalgal bioremediation of petroleum-derived low salinity and low pH produced water. Journal of Applied Phycology, 31, 435-444. https://doi.org/10.1007/s10811-018-1571-6
de-Bashan, L. E. y Bashan, Y. (2010). Immobilized microalgae for removing pollutants: review of practical aspects. Bioresource Technology, 101(6), 1611-1627. https://doi.org/10.1016/j.biortech.2009.09.043
Díaz-Borrego, L., Vera, A., Marín, J., Aiello, C., Briceño, B. y Morales, E. (2012). Efecto del queroseno y de la concentración de nutrientes en el crecimiento de un cultivo mixto de microalgas (Chlorophyta). Revista de la Universidad del Zulia 3ra Época, 3(6), 102-118. https://www.produccioncientifica.luz.edu.ve/index.php/rluz/article/view/31185
Dubois, M., Guilles, K. A., Hamilton, J. K., Rebers, P. A. y Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 28(3), 350-356. https://doi.org/10.1021/ac60111a017
Egberomoh, G. O. y Fagade, O. E. (2016). Microalgal-bacterial consortium in polyaromatic hydrocarbon degradation of petroleum – based effluent. Journal of Bioremediation & Biodegradation, 7, 359. https://doi.org/10.4172/2155-6199.1000359
El-Sayed-Touliabah, H., El-Sheekh, M. M., Ismail, M. M. y El-Kassas, H. (2022). A review of microalgae- and cyanobacteria-based biodegradation of organic pollutants. Molecules, 27, 1141. https://doi.org/10.3390/molecules27031141
Fabregas, J., Herrero, C. y Veiga, M. (1984). Effect of oil and dispersant on growth and chlorophyll α content of the marine microalga Tetraselmis suecica. Applied and Environmental Microbiology, 47(2), 445-447. https://doi.org/10.1128/aem.47.2.445-447.1984
Gonzalez-Gonzalez, L. M. y de-Bashan, L. E. (2023). The potential of microalgae-bacteria consortia to restore degraded soils. Biology, 12(5), 693. https://doi.org/10.3390/biology12050693
Guldhe, A., Ansari, F. A., Singh, P. y Bux, F. (2017). Heterotrophic cultivation of microalgae using aquaculture wastewater: A biorefinery concept for biomass production and nutrient remediation. Ecological Engineering, 99, 47-53. https://doi.org/10.1016/j.ecoleng.2016.11.013
Hachicha, R., Elleuch, F., Ben-Hlima, H., Dubessay, P., de Baynast, H., Delattre, C., Pierre, G., Hachicha, R., Abdelkafi, S., Michaud, P. y Fendri, I. (2022). Biomolecules from microalgae and cyanobacteria: applications and market survey. Applied Sciences, 12, 1924. https://doi.org/10.3390/app12041924
Hammed, A. M., Prajapati, S. K., Simsek, S. y Simsek, H. (2016). Growth regime and environmental remediation of microalgae. Algae, 31(3), 189-204. http://dx.doi.org/10.4490/algae.2016.31.8.28
Hamouda, R. A., Alhumairi, A. M. y Saddiq, A. A. (2023). Simultaneous bioremediation of petroleum hydrocarbons and production of biofuels by the micro-green alga, cyanobacteria, and its consortium. Heliyon, 9, e16656. https://doi.org/10.1016/j.heliyon.2023.e16656
Hamouda, R. A., Sorour, N. M. y Yeheia, D. S. (2016). Biodegradation of crude oil by Anabaena oryzae, Chlorella kessleri and its consortium under mixotrophic conditions. International Biodeterioration & Biodegradation, 112, 128-134. https://doi.org/10.1016/j.ibiod.2016.05.001
Hanif, A., Shah, F. H., Mumtaz, A. S., Ullah, T., Azeem, M. A., Ahmad, M. y Shah, A. A. (2024). Taxonomic study of freshwater microalgal diversity and its optimum culturing condition of District Karak, Pakistan. Journal of Biodiversity and Environmental Sciences, 24(3), 96-116. https://www.innspub.net/wp-content/uploads/2024/06/JBES-V24-No3-p97-116.pdf
Hebert, D., Phipps, P. J. y Strange, R. E. (1971). Chemical analysis of microbial cells. Methods in Microbiology, 5, 209-344. http://dx.doi.org/10.1016/S0580-9517(08)70641-X
Hossain, M. F., Akter, M. A., Sohan, M. S. R., Sultana, D. N., Reza, M. A. y Hoque, K. M. F. (2022). Bioremediation potential of hydrocarbon degrading bacteria: isolation, characterization, and assessment. Saudi Journal of Biological Sciences, 29(1), 211-216. https://doi.org/10.1016/j.sjbs.2021.08.069
Jeffrey, S. W. y Humphrey, G. F. (1975). New spectrophotometric equations for determining chlorophylls a, b, c1 and c2 in higher plants, algae and natural phytoplankton. Biochemie und Physiologie der Pflanzen, 167(2), 191-194. http://doi.org/10.1016/S0015-3796(17)30778-3
Ji, Y., Hu, W., Li, X., Ma, G., Song, M. y Pei, H. (2014). Mixotrophic growth and biochemical analysis of Chlorella vulgaris cultivated with diluted monosodium glutamate wastewater. Bioresource Technology, 152, 471-476. https://doi.org/10.1016/j.biortech.2013.11.047
Jiménez-Guanipa, H. y Viedma, E. (2018). Energía, cambio climático y desarrollo sostenible. Impacto sobre los derechos humanos. Fundación Heinrich Böll.
Khan, Z. I., Ahmad, K., Siddique, S., Ahmad, T., Bashir, H., Munir, M., Mahpara, S., Malik, I. S., Wajid, K., Ugulu, I., Nadeem, M., Noorka, I. R. y Chen, F. (2020). A study on the transfer of chromium from meadows to grazing livestock: an assessment of health risk. Environmental Science and Pollution Research International, 27(21), 26694-26701. https://doi.org/10.1007/s11356-020-10660-z
Kochert, G. (1978). Carbohydrate determination by the phenol-sulfuric acid method. En: J. Hellebust y J. Craigie (Eds.). Handbook of Phycological Methods. Physiological and Biochemical Methods. Cambridge University Press.
Lee, J., Hong, S., An, S. A. y Khim, J. S. (2023). Methodological advances and future directions of microalgal bioassays for evaluation of potential toxicity in environmental samples: a review. Environment International, 173, 107869. https://doi.org/10.1016/j.envint.2023.107869
León-Vaz, A., Torres-Franco, A. F., García-Encina, P. A. y Muñoz, R. (2025). Developing a microalgal-bacterial consortium for the removal of organic pollutants from petrochemical industry. Journal of Water Process Engineering, 73, 107663. https://doi.org/10.1016/j.jwpe.2025.107663
Lobban, C., Chapman, D. y Kremer, B. (1988). Experimental phycology: a laboratory manual. Cambridge University Press.
Lopes da Silva, T., Moniz, P., Silva, C. y Reis, A. (2021). The role of heterotrophic microalgae in waste conversion to biofuels and bioproducts. Processes, 9, 1090. https://doi.org/10.3390/pr9071090
López, J., Quintero, G., Guevara, A., Jaimes, D., Gutiérrez, S. y García, J. (2006). Biorremediación de suelos contaminados con hidrocarburos derivados del petróleo. Nova, 4(5), 82-90. https://doi.org/10.22490/24629448.351
Lowry, H., Rosebrough, J., Farr, L. y Randall, J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265-275. https://pubmed.ncbi.nlm.nih.gov/14907713/
Lucana, N. y Huanca, R. (2014). Estructura bacteriana. Revista de Actualización Clínica, 49, 2589- 2593. http://revistasbolivianas.umsa.bo/pdf/raci/v49/v49_a01.pdf
Madigan, M. T., Martinko, J. M., Bender, K. S., Buckley, D. H. y Stahl, D. A. (2015). Brock, Biología de microorganismos. (14ta edición). Pearson Educación S. A.
Manasseh, I. M. y Humphrey, S. S. (2024). Bioremediation of oil spill: concept, methods and applications. Discover Chemistry, 1, 42. https://doi.org/10.1007/s44371-024-00038-2
Marsh, J. y Weinstein, D. (1966). Simple charring method for determination of lipids. Journal of Lipid Research, 7(4), 574-576. https://doi.org/10.1016/S0022-2275(20)39274-9
Miazek K., Kratky, L., Sulc, R., Jirout, T., Aguedo, M., Richel, A. y Goffin, D. (2017). Effect of organic solvents on microalgae growth, metabolism and industrial bioproduct extraction: a review. International Journal of Molecular Sciences, 18, 1429. https://doi.org/10.3390/ijms18071429
Mona, S., Kaushik, A. y Kaushik, C. P. (2011). Biosorption of reactive dye by waste biomass of Nostoc linckia. Ecological Engineering, 37(10), 1589-1594. https://doi.org/10.1016/j.ecoleng.2011.04.005
Muñoz-Peñuela, M., Ramírez, J., Otero-Paternina, A. M., Medina-Robles, V. M., Cruz-Casallas, P. E. y Velasco-Santamaría, V. (2012). Effect of culture medium on growth and protein content of Chlorella vulgaris. Revista Colombiana de Ciencias Pecuarias, 25(3), 438-449. http://www.scielo.org.co/pdf/rccp/v25n3/v25n3a12.pdf
Needham, J. y Needham, P. R. (1978). Guía de estudio de los seres vivos de las aguas dulces. Editorial Reverté, S. A.
Okeke, E. S., Okoye, C. O., Chidike-Ezeorba, T. P., Mao, G., Chen, Y., Xu, H., Song, C., Feng, W. y Wu, X. (2022). Emerging bio-dispersant and bioremediation technologies as environmentally friendly management responses toward marine oil spill: a comprehensive review. Journal of Environmental Management, 322, 116123. https://doi.org/10.1016/j.jenvman.2022.116123
Otero-Paternina, A., Cruz-Casallas, P. E. y Velasco-Santamaría, Y. M. (2013). Evaluación del efecto del hidrocarburo fenantreno sobre el crecimiento de Chlorella vulgaris (Chlorellaceae). Acta Biológica Colombiana, 18(1), 87-98. https://revistas.unal.edu.co/index.php/actabiol/article/view/24558/39935
Patowary, K., Patowary, R., Kalita, M. C. y Deka, S. (2016). Development of an efficient bacterial consortium for the potential remediation of hydrocarbons from contaminated sites. Frontiers in Microbiology, 14(7), 1092. https://doi.org/10.3389/fmicb.2016.01092
Proietti-Tocca, G., Agostino, V., Menin, B., Tommasi, T., Fino, D. y Di Caprio, F. (2024). Mixotrophic and heterotrophic growth of microalgae using acetate from different production processes. Reviews in Environmental Science and Bio/Technology, 23, 93-132. https://doi.org/10.1007/s11157-024-09682-7
Radice, R. P., De Fabrizio, V., Donadoni, A., Scopa, A. y Martelli, G. (2023). Crude oil bioremediation: from bacteria to microalgae. Processes, 11, 442. https://doi.org/10.3390/pr11020442
Rodriguez, K. N. D., Santos, R. T., Nagpala, M. J. M. y Opulencia, R. B. (2023). Metataxonomic characterization of enriched consortia derived from oil spill-contaminated sites in Guimaras, Philippines, reveals major role of Klebsiella sp. in hydrocarbon degradation. International Journal of Microbiology, 2023, 3247448. https://doi.org/10.1155/2023/3247448
Roy, U. K., Wagner, J. y Radu, T. (2023). Production of metabolites in microalgae under alkali halophilic growth medium using a dissolved inorganic carbon source. Waste and Biomass Valorization, 14, 3339-3354. https://doi.org/10.1007/s12649-023-02053-3
Sánchez, J. C. (2021). Afectación de los ecosistemas marino-costeros por los derrames de hidrocarburos. Boletín de la Academia de Ciencias Físicas, Matemáticas y Naturales, LXXXI(1), 35-39. https://acfiman.org/wp-content/uploads/2022/07/LXXXI.N1.P35-39.2021.pdf
Satpati, G. G., Gupta, S., Biswas, R. K., Choudhury, A. K., Kim, J. W. y Davoodbasha, M. A. (2023). Microalgae mediated bioremediation of polycyclic aromatic hydrocarbons: strategies, advancement and regulations. Chemosphere, 344, 140337. https://doi.org/10.1016/j.chemosphere.2023.140337
Schmitt, D., Müller, A., Csögör, Z., Frimmel, F. H. y Posten, C. (2001). The adsorption kinetics of metal ions onto different microalgae and siliceous earth. Water Research, 35(3), 779-785. https://doi.org/10.1016/S0043-1354(00)00317-1
Silva, D. A., Cardoso, L. G., Silva, J. S., Oliveira de Souza, C., França-Lemos, P. V., de Almeida, P. F., de Souza-Ferreira, E., Lombardi, A. T. y Druzian, J. I. (2022). Strategy for the cultivation of Chlorella vulgaris with high biomass production and biofuel potential in wastewater from the oil industry. Environmental Technology & Innovation, 25, 102204. https://doi.org/10.1016/j.eti.2021.102204
Strickland, J. y Parsons, T. (1972). A practical handbook of seawater analysis. (2nd edition). Fisheries Research Board Canadian.
Vo, T. P., Danaee, S., Chaiwong, C., Pham, B. T., Poddar, N., Kim, M., Kuzhiumparambil, U., Songsomboon, C., Pernice, M., Ngo, H. H., Ralph, P. J. y Vo, P. H. N. (2024). Microalgae-bacteria consortia for organic pollutants remediation from wastewater: a critical review. Journal of Environmental Chemical Engineering, (6)12, 114213. https://doi.org/10.1016/j.jece.2024.114213
Wei, X., Peng, P., Meng, Y., Li, T., Fan, Z., Wang, Q. y Chen, J. (2021). Degradation performance of petroleum-hydrocarbon-degrading bacteria and its application in remediation of oil contaminated soil. IOP Conference Series: Earth and Environmental Science, 766, 012096. https://doi.org/10.1088/1755-1315/766/1/012096
Wilkes, H., Jarling, R. y Schwarzbauer, J. (2020). Hydrocarbons and lipids: an introduction to structure, physicochemical properties, and natural occurrence. En: H. Wilkes (Ed.). Hydrocarbons, oils and lipids: diversity, origin, chemistry and fate. Handbook of hydrocarbon and lipid microbiology. Springer Nature. https://doi.org/10.1007/978-3-319-90569-3_34
Wu, B., Xiu, J., Yu, L., Huang, L., Yi, L. y Ma, Y. (2023). Degradation of crude oil in a co-culture system of Bacillus subtilis and Pseudomonas aeruginosa. Frontiers in Microbiology, 14, 1132831. https://doi.org/10.3389/fmicb.2023.1132831
Yacubson, S. (1969). Algas de ambientes acuáticos continentales, nuevas para Venezuela (Cyanophyta, Chlorophyta). Boletín del Centro de Investigaciones Biológicas, 3, 1-87. https://produccioncientificaluz.org/index.php/boletin/article/view/168
Yamaguchi, T., Ishida, M. y Suzuki, T. (1999). Biodegradation of hydrocarbons by Prototheca zopfii in rotating biological contactors. Process Biochemistry, 35, 403-409. https://doi.org/10.1016/S0032-9592(99)00086-2
