Rapid Colonization of Uranium Mining-Impacted Waters, the Biodiversity of Successful Lineages of Phytoplankton Extremophiles

  • Beatriz Baselga-Cervera
  • Camino García-BalboaEmail author
  • Héctor M. Díaz-Alejo
  • Eduardo Costas
  • Victoria López-Rodas
Environmental Microbiology


Anthropogenic extreme environments are emphasized as interesting sites for the study of evolutionary pathways, biodiversity, and extremophile bioprospection. Organisms that grow under these conditions are usually regarded as extremophiles; however, the extreme novelty of these environments may have favor adaptive radiations of facultative extremophiles. At the Iberian Peninsula, uranium mining operations have rendered highly polluted extreme environments in multiple locations. In this study, we examined the phytoplankton diversity, community structure, and possible determining factors in separate uranium mining-impacted waters. Some of these human-induced extreme environments may be able to sustain indigenous facultative extremophile phytoplankton species, as well as alleged obligate extremophiles. Therefore, we investigated the adaptation capacity of three laboratory strains, two Chlamydomonas reinhardtii and a Dictyosphaerium chlorelloides, to uranium-polluted waters. The biodiversity among the sampled waters was very low, and despite presenting unique taxonomic records, ecological patterns can be identified. The microalgae adaptation experiments indicated a gradient of ecological novelty and different phenomena of adaptation, from acclimation in some waters to non-adaptation in the harshest anthropogenic environment. Certainly, phytoplankton extremophiles might have been often overlooked, and the ability to flourish in extreme environments might be a functional feature in some neutrophilic species. Evolutionary biology and microbial biodiversity can benefit the study of recently evolved systems such as uranium-polluted waters. Moreover, anthropogenic extremophiles can be harnessed for industrial applications.


Uranium mining impacted waterbodies Phytoplankton Anthropogenic extreme environments Microbial biodiversity Facultative extremophiles Adaptation 



uranium mining impacted water bodies





ENUSA was instrumental in facilitating sampling. Pedro Caravantes, Antonio García-Sanchez, and José Mariano Igual provided invaluable assistance with sampling and chemical analyses. Thanks are given to Lara de Miguel Fernandez for her excellent technical support.

Authors’ Contributions

BB-C, CG-B, EC, and VL-R conceived and planned the experiments. BB-C and CG-B interpreted the results and worked in the manuscript, with a special interest in the conceptual model. BB-C and CG-B carried out sampling and experimentation. HMD aided in the interpretation of the results. HMD and EC performed the statistical analyses. BB-C wrote the manuscript with inputs from all authors. CG-G, EC, and VL-R supervised the project.

Funding Information

This study is supported by the Spanish Secretaría de Estado, Investigación, Desarrollo e Innovación, grant CTM-2013-44366-R.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

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High resolution image (TIF 13126 kb)


  1. 1.
    Martiny JBH, Bohannan BJM, Brown JH, Colwell RK, Fuhrman JA, Green JL, Horner-Devine MC, Kane M, Krumins JA, Kuske CR, Morin PJ, Naeem S, Øvreås L, Reysenbach AL, Smith VH, Staley JT (2006) Microbial biogeography: putting microorganisms on the map. Nat Rev Microbiol 4:102–112. CrossRefGoogle Scholar
  2. 2.
    Rothschild LJ (2001) Life in extreme environments. Ad Astra 14:1092–1101. Google Scholar
  3. 3.
    Anitori RP (2012) Extremophiles: microbiology and biotechnology. Casiter Academic Press, Norfolk, UKGoogle Scholar
  4. 4.
    Fuciños P, González R, Atanes E et al (2012) Lipases and esterases’ from extremophiles: overview and case example of the production and purification of an esterase from Thermus thermophilus HB27. Methods Mol Biol 861:239–266. CrossRefGoogle Scholar
  5. 5.
    Low-Décarie E, Fussmann GF, Dumbrell AJ, Bell G (2016) Communities that thrive in extreme conditions captured from a freshwater lake. Biol Lett 12:20160562. CrossRefGoogle Scholar
  6. 6.
    Usami R, Echigo A, Fukushima T, Mizuki T, Yoshida Y, Kamekura M (2007) Alkalibacillus silvisoli sp. nov., an alkaliphilic moderate halophile isolated from non-saline forest soil in Japan. Int J Syst Evol Microbiol 57:770–774. CrossRefGoogle Scholar
  7. 7.
    Gibbons SM, Caporaso JG, Pirrung M, Field D, Knight R, Gilbert JA (2013) Evidence for a persistent microbial seed bank throughout the global ocean. Proc Natl Acad Sci 110:4651–4655. CrossRefGoogle Scholar
  8. 8.
    Finlay BJ (2002) Global dispersal of free-living microbial eukaryote species. Science 296:1061–1063. CrossRefGoogle Scholar
  9. 9.
    Baas-Becking L (1934) Geobiologie; of inleiding tot de milieukundeGoogle Scholar
  10. 10.
    Rothschild LJ, Mancinelli RL (2001) Life in extreme environments. Nature 409:1092–1101. CrossRefGoogle Scholar
  11. 11.
    Varshney P, Mikulic P, Vonshak A, Beardall J, Wangikar PP (2015) Extremophilic micro-algae and their potential contribution in biotechnology. Bioresour Technol 184:363–372. CrossRefGoogle Scholar
  12. 12.
    Seckbach J (2007) Algae and cyanobacteria in extreme environments. Springer, DordrechtCrossRefGoogle Scholar
  13. 13.
    Denicola DM (2000) A review of diatoms found in highly acidic environments. Hydrobiologia 433:111–122CrossRefGoogle Scholar
  14. 14.
    Costas E, Flores-Moya A, Perdigones N, Maneiro E, Blanco JL, García ME, López-Rodas V (2007) How eukaryotic algae can adapt to the Spain’s Rio Tinto: a neo-Darwinian proposal for rapid adaptation to an extremely hostile ecosystem. New Phytol 175:334–339. CrossRefGoogle Scholar
  15. 15.
    Amaral Zettler LA, Gómez F, Zettler E, Keenan BG, Amils R, Sogin ML (2002) Microbiology: eukaryotic diversity in Spain’s river of fire. Nature 417:137–137. CrossRefGoogle Scholar
  16. 16.
    Markich SJ (2002) Uranium speciation and bioavailability in aquatic systems: an overview. Sci World J 2:707–729. CrossRefGoogle Scholar
  17. 17.
    Kim J-I, Buckau G, Klenze R (1992) The chemical behavior of transuranium elements and barrier functions in natural aquifer systems. MRS Proc 294:3. CrossRefGoogle Scholar
  18. 18.
    Sattler B, Post B, D. R, et al (2012) Life at extremes: environments, organisms, and strategies for survival. 554.
  19. 19.
    Zirnstein I, Arnold T, Krawczyk-Bärsch E, Jenk U, Bernhard G, Röske I (2012) Eukaryotic life in biofilms formed in a uranium mine. Microbiologyopen 1:83–94. CrossRefGoogle Scholar
  20. 20.
    Schippers A, Hallmann R, Wentzien S, Sand W (1995) Microbial diversity in uranium mine waste heaps. Appl Environ Microbiol 61:2930–2935Google Scholar
  21. 21.
    Nevin KP, Finneran KT, Lovley DR (2003) Microorganisms associated with uranium bioremediation in a high-salinity subsurface sediment. Appl Environ Microbiol 69:3672–3675. CrossRefGoogle Scholar
  22. 22.
    Campos MB, de Azevedo H, Nascimento MRL, Roque CV, Rodgher S (2011) Environmental assessment of water from a uranium mine (Caldas, Minas Gerais state, Brazil) in a decommissioning operation. Environ Earth Sci 62:857–863. CrossRefGoogle Scholar
  23. 23.
    García-Balboa C, Baselga-Cervera B, García-Sanchez A, Igual JM, Lopez-Rodas V, Costas E (2013) Rapid adaptation of microalgae to bodies of water with extreme pollution from uranium mining: an explanation of how mesophilic organisms can rapidly colonise extremely toxic environments. Aquat Toxicol 144–145:116–123. CrossRefGoogle Scholar
  24. 24.
    Baselga-Cervera B, Romero-López J, García-Balboa C et al (2018) Improvement of the uranium sequestration ability of a Chlamydomonas sp. (ChlSP strain) isolated from extreme uranium mine tailings through selection for potential bioremediation application. Front Microbiol 9:523. CrossRefGoogle Scholar
  25. 25.
    Baselga-Cervera B, López-Rodas V, García-Balboa C, Costas E (2013) Microalgae : the first nuclear engineers ? An Real Acad Farm 79:634–645Google Scholar
  26. 26.
    Bellinger EG, Sigee DC (2015) Freshwater algae: identification and use as bioindicators. John Wiley & Sons, Ltd, Chichester, UKGoogle Scholar
  27. 27.
    Kimura M, Maruyama T (1966) The mutational load with epistatic gene interactions in fitness. Genetics 54:213–220Google Scholar
  28. 28.
    Novick A, Szilard L (1950) Experiments with the Chemostat on spontaneous mutations of bacteria. Proc Natl Acad Sci U S A 36:708–719CrossRefGoogle Scholar
  29. 29.
    Dykhuizen DE, Hartl DL (1983) Selection in chemostats. Microbiol Rev 47:150–168Google Scholar
  30. 30.
    Wortel MT, Bosdriesz E, Teusink B, Bruggeman FJ (2016) Evolutionary pressures on microbial metabolic strategies in the chemostat. Sci Rep 6:29503. CrossRefGoogle Scholar
  31. 31.
    Goho S, Bell G (2000) The ecology and genetics of fitness in Chlamydomonas. IX. The rate of accumulation of variation of fitness under selection. Evolution 54:416–424CrossRefGoogle Scholar
  32. 32.
    Kliphuis AMJ, Klok AJ, Martens DE, Lamers PP, Janssen M, Wijffels RH (2012) Metabolic modeling of Chlamydomonas reinhardtii: energy requirements for photoautotrophic growth and maintenance. J Appl Phycol 24:253–266. CrossRefGoogle Scholar
  33. 33.
    Ferenci T (2007) Bacterial physiology, regulation and mutational adaptation in a Chemostat environment. Adv Microb Physiol 53:169–315. CrossRefGoogle Scholar
  34. 34.
    Stent G, Calendar R (1978) Molecular genetics. Free San FrGoogle Scholar
  35. 35.
    Williams M (1977) Sterological techniques. In: Glanert AM (ed) Practical methods in electron microscopy. North Holland/America Elsevier, Amsterdam, pp 1–216Google Scholar
  36. 36.
    Weibel ER, Bolender R (1973) Stereological techniques for electron microscopic morphometry. In: Hayat M (ed) Principles and techniques of electron microscopy. Van Nostrand Reynold CO, Nwe York, pp 237–293Google Scholar
  37. 37.
    World Nuclear Association (2018) Occupational Safety in Uranium Mining - World Nuclear Association. Accessed 21 Mar 2019
  38. 38.
    Shannon C (1948) A mathematical theory of communication. Bell Syst Tech J 27:379–423CrossRefGoogle Scholar
  39. 39.
    Simpson EH (1949) Measurement of diversity. Nature 163:688–688CrossRefGoogle Scholar
  40. 40.
    Pal R, Choudhury AK (2014) An introduction to phytoplanktons: diversity and ecologyGoogle Scholar
  41. 41.
    Wolowski K, Turnau K, Henriques FS (2008) The algal flora of an extremely acidic, metal-rich drainage pond of Sao Domingos pyrite mine (Portugal). Cryptogam Algol 29:313–324Google Scholar
  42. 42.
    López-Rodas V, Marvá F, Rouco M, Costas E, Flores-Moya A (2008) Adaptation of the chlorophycean Dictyosphaerium chlorelloides to stressful acidic, mine metal-rich waters as result of pre-selective mutations. Chemosphere 72:703–707. CrossRefGoogle Scholar
  43. 43.
    Płachno BJ, Wołowski K, Augustynowicz J, Łukaszek M (2015) Diversity of algae in a thallium and other heavy metals-polluted environment. Ann Limnol Int J Limnol 51:139–146. CrossRefGoogle Scholar
  44. 44.
    Cao Y, Kalin M (1999) Phytoplankton in mine waste water community structure, control factors and biological monitoring. Nat Resour Canada 240Google Scholar
  45. 45.
    Reed RH, Gadd GM (1990) Metal tolerance in eukaryotic and prokaryotic algaeGoogle Scholar
  46. 46.
    Rai LC, Gaur JP, Kumar HD (1981) Phycology and heavy-metal pollution. Biol Rev 56:99–151. CrossRefGoogle Scholar
  47. 47.
    Steinberg CEW, Schäfer H, Beisker W (1998) Do acid-tolerant cyanobacteria exist? Acta Hydrochim Hydrobiol 26:13–19.<13::AID-AHEH13>3.0.CO;2-V CrossRefGoogle Scholar
  48. 48.
    Nancucheo I, Barrie Johnson D (2012) Acidophilic algae isolated from mine-impacted environments and their roles in sustaining heterotrophic acidophiles. Front Microbiol 3:325. CrossRefGoogle Scholar
  49. 49.
    Pick U (1999) Dunaliella Acidophila — A Most Extreme Acidophilic Alga. Enigmatic microorganisms and life in extreme environments. Springer Netherlands, Dordrecht, pp 465–478CrossRefGoogle Scholar
  50. 50.
    Peterson HG, Nyholm N, Nelson M et al (1996) Development of aquatic plant bioassays for rapid screening and interpretive risk assessments of metal mining liquid waste waters. Water Science and Technology, pp 155–161Google Scholar
  51. 51.
    Peterson HG, Healey FP, Wagemann R (1984) Metal toxicity to algae: a highly pH dependent phenomenon. Can J Fish Aquat Sci 41:974–979. CrossRefGoogle Scholar
  52. 52.
    Maeda S, Kusadome K, Arima H, Ohki A, Naka K (1992) Biomethylation of arsenic and its excretion by the algaChlorella vulgaris. Appl Organomet Chem 6:407–413. CrossRefGoogle Scholar
  53. 53.
    Cullen WR, Harrison LG, Li H, Hewitt G (1994) Bioaccumulation and excretion of arsenic compounds by a marine unicellular alga,polyphysa peniculus. Appl Organomet Chem 8:313–324. CrossRefGoogle Scholar
  54. 54.
    Wang Y, Wang S, Xu P, Liu C, Liu M, Wang Y, Wang C, Zhang C, Ge Y (2015) Review of arsenic speciation, toxicity and metabolism in microalgae. Rev Environ Sci Biotechnol/Technol 14:427–451. CrossRefGoogle Scholar
  55. 55.
    Levy JL, Stauber JL, Adams MS, Maher WA, Kirby JK, Jolley DF (2005) Toxicity, biotransformation, and mode of action of ARSENIC in two freshwater microalgae (Chlorella sp. and Monoraphidium arcuatum). Environ Toxicol Chem 24:2630–2639. CrossRefGoogle Scholar
  56. 56.
    Vocke RW, Sears KL, O’Toole JJ, Wildman RB (1980) Growth responses of selected freshwater algae to trace elements and scrubber ash slurry generated by coal-fired power plants. Water Res 14:141–150. CrossRefGoogle Scholar
  57. 57.
    Baos R, García-Villada L, Agrelo M, López-Rodas V, Hiraldo F, Costas E (2002) Short-term adaptation of microalgae in highly stressful environments: an experimental model analysing the resistance of Scenedesmus intermedius (Chlorophyceae) to the heavy metals mixture from the Aznalcóllar mine spill. Eur J Phycol 37:593–600. CrossRefGoogle Scholar
  58. 58.
    Franklin NM, Stauber JL, Markich SJ, Lim RP (2000) pH-dependent toxicity of copper and uranium to a tropical freshwater alga (Chlorella sp.). Aquat Toxicol 48:275–289. CrossRefGoogle Scholar
  59. 59.
    De Schamphelaere KAC, Stauber JL, Wilde KL et al (2005) Toward a biotic ligand model for freshwater green algae: surface-bound and internal copper are better predictors of toxicity than free Cu2+-ion activity when pH is varied. Environ Sci Technol 39:2067–2072. CrossRefGoogle Scholar
  60. 60.
    Wilde KL, Stauber JL, Markich SJ, Franklin NM, Brown PL (2006) The effect of pH on the uptake and toxicity of copper and zinc in a tropical freshwater alga (Chlorella sp.). Arch Environ Contam Toxicol 51:174–185. CrossRefGoogle Scholar
  61. 61.
    Goulet RR, Thompson PA, Serben KC, Eickhoff CV (2015) Impact of environmentally based chemical hardness on uranium speciation and toxicity in six aquatic species. Environ Toxicol Chem 34:562–574. CrossRefGoogle Scholar
  62. 62.
    Fortin C, Dutel L, Garnier-Laplace J (2004) Uranium complexation and uptake by a green alga in relation to chemical speciation: the importance of the free uranyl ion. Environ Toxicol Chem 23:974–981. CrossRefGoogle Scholar
  63. 63.
    Lavoie M, Sabatier S, Garnier-Laplace J, Fortin C (2014) Uranium accumulation and toxicity in the green alga Chlamydomonas reinhardtii is modulated by pH. Environ Toxicol Chem 33:1372–1379. CrossRefGoogle Scholar
  64. 64.
    Herlory O, Bonzom J-M, Gilbin R (2013) Sensitivity evaluation of the green alga Chlamydomonas reinhardtii to uranium by pulse amplitude modulated (PAM) fluorometry. Aquat Toxicol 140–141:288–294. CrossRefGoogle Scholar
  65. 65.
    Charles AL, Markich SJ, Stauber JL, De Filippis LF (2002) The effect of water hardness on the toxicity of uranium to a tropical freshwater alga Chlorella sp. Aquat Toxicol 60:61–73CrossRefGoogle Scholar
  66. 66.
    Chankova SG, Bryant PE (2002) Acceleration of DNA-double strand rejoining during the adaptive response of Chlamydomonas reinhardtii. Radiats Biol Radioecol 42:600–603Google Scholar
  67. 67.
    Rea G, Esposito D, Damasso M, Serafini A, Margonelli A, Faraloni C, Torzillo G, Zanini A, Bertalan I, Johanningmeier U, Giardi MT (2008) Ionizing radiation impacts photochemical quantum yield and oxygen evolution activity of photosystem II in photosynthetic microorganisms. Int J Radiat Biol 84:867–877. CrossRefGoogle Scholar
  68. 68.
    Gomes T, Xie L, Brede D, Lind OC, Solhaug KA, Salbu B, Tollefsen KE (2017) Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: photosynthetic performance and ROS formation. Aquat Toxicol 183:1–10. CrossRefGoogle Scholar
  69. 69.
    Novis PM, Harding JS (2007) Extreme Acidophiles. In: Seckbach J (ed) Algae and cyanobacteria in extreme environments. Springer, Dordrecht, pp 443–463CrossRefGoogle Scholar
  70. 70.
    Gould SJ (2002) The structure of evolutionary theory. Belknap Press of Harvard University PressGoogle Scholar
  71. 71.
    López-Rodas V, Marvá F, Costas E, Flores-Moya A (2008) Microalgal adaptation to a stressful environment (acidic, metal-rich mine waters) could be due to selection of pre-selective mutants originating in non-extreme environments. Environ Exp Bot 64:43–48. CrossRefGoogle Scholar
  72. 72.
    Flores-Moya A, Costas E, Bañares-España E, García-Villada L, Altamirano M, López-Rodas V (2005) Adaptation of Spirogyra insignis (Chlorophyta) to an extreme natural environment (sulphureous waters) through preselective mutations. New Phytol 166:655–661. CrossRefGoogle Scholar
  73. 73.
    Cervera BB, Rodas VL, Balboa G (2016) Mechanisms of rapid adaptation to environmental stressors in phytoplankton. J Environ Anal Toxicol 6.
  74. 74.
    Sniegowski PD (2005) Linking mutation to adaptation: overcoming stress at the spa. New Phytol 166:360–362. CrossRefGoogle Scholar
  75. 75.
    Souza-Egipsy V, Altamirano M, Amils R, Aguilera A (2011) Photosynthetic performance of phototrophic biofilms in extreme acidic environments. Environ Microbiol 13:2351–2358. CrossRefGoogle Scholar
  76. 76.
    Rowe OF, Sánchez-España J, Hallberg KB, Johnson DB (2007) Microbial communities and geochemical dynamics in an extremely acidic, metal-rich stream at an abandoned sulfide mine (Huelva, Spain) underpinned by two functional primary production systems. Environ Microbiol 9:1761–1771. CrossRefGoogle Scholar
  77. 77.
    Valente T, Gomes CL (2007) The role of two acidophilic algae as ecological indicators of acid mine drainage sites.pdf. J Iber Geol 33:283–294Google Scholar
  78. 78.
    Gimmler H (2001) Acidophilic and acidotolerant algae. Algal adaptation to environmental stresses. Springer, Berlin Heidelberg, pp 259–290CrossRefGoogle Scholar
  79. 79.
    Seckbach J, Oren A (2007) Oxygenic photosynthetic microorganisms in extreme environments. Springer, Dordrecht, pp 3–25Google Scholar
  80. 80.
    Impey C (2007) The living Cosmos: our search for life in the universeGoogle Scholar
  81. 81.
    Li S-J, Hua Z-S, Huang L-N, Li J, Shi SH, Chen LX, Kuang JL, Liu J, Hu M, Shu WS (2015) Microbial communities evolve faster in extreme environments. Sci Rep 4:6205. CrossRefGoogle Scholar
  82. 82.
    Bowers KJ, Mesbah NM, Wiegel J (2009) Biodiversity of poly-extremophilic bacteria: does combining the extremes of high salt, alkaline pH and elevated temperature approach a physico-chemical boundary for life? Saline Syst 5:9. CrossRefGoogle Scholar
  83. 83.
    García-Meza JV, Barrangue C, Admiraal W (2005) Biofilm formation by algae as a mechanism for surviving on mine tailings. Environ Toxicol Chem 24:573–581. CrossRefGoogle Scholar
  84. 84.
    Wilde EW, Benemann JR (1993) Bioremoval of heavy metals by the use of microalgae. Biotechnol Adv 11:781–812CrossRefGoogle Scholar
  85. 85.
    Kalin M, Wheeler WN, Meinrath G (2004) The removal of uranium from mining waste water using algal/microbial biomass. J Environ Radioact 78:151–177. CrossRefGoogle Scholar
  86. 86.
    Baselga-Cervera B, García-Balboa C, López-Rodas V, Fernández Díaz M, Costas E (2019) Evidence of microalgal isotopic fractionation through enrichment of depleted uranium. Sci Rep 9:1973. CrossRefGoogle Scholar

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Authors and Affiliations

  1. 1.Animal Science (Genetics), School of Veterinary MedicineComplutense University of MadridMadridSpain
  2. 2.Ecology, Evolution and Behavior DepartmentUniversity of MinnesotaSt. PaulUSA

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