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Specific detection of bioavailable phenanthrene and mercury by bacterium reporters in the red soil

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Abstract

Genetically engineered Pseudomonas putida reporters (BMB-PL and BMB-ME), which, respectively, carried phnS-luxCDABE and merR-egfp cassette, were used to determine bioavailable phenanthrene and mercury. Over a spiked range of concentrations and aged for 6 days in red soil samples, the reporters were tested to determine the optimal assay conditions and the bioavailable phenanthrene (0–60 mg kg−1) and Hg2+ (0–240 μg kg−1) were evaluated by the signal of the relative fluorescent units and relative luminescence units. Single contamination was carried out and good correlations were obtained between signal strength and pollutant concentrations, whereas interference and bioavailability repression were observed in dual-contamination experiments. Other heavy metal ions at nanomolar level did not interfere with BMB-ME measurement while BMB-PL showed some response to other polycyclic aromatic hydrocarbons or their intermediate products during degradation. Comparing high-performance liquid chromatography methods with the bacterial reporters, both BMB-ME and BMB-PL appeared to have a detection limit (mercury <40 μg kg−1; phenanthrene <24 mg kg −1) similar to the instrumental analysis. Although physical parameters may affect the interaction of pollutants with bioreporter cells, advantages include the inherent biological relevance of the response, rapid response time, and potential for field deployment. Our results strongly suggest that the BMB-ME and BMB-PL bioreporters constitute an adaptable system for easily detecting the bioavailability of mercury and phenanthrene in the red soils.

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References

  • Applegate BM, Kehrmeyer SR, Sayler GS (1998) A chromosomally based tod-luxCDABE whole-cell reporter for benzene, toluene, ethybenzene, and xylene (BTEX) sensing. Appl Environ Microbiol 64(7):2730–2735

    CAS  Google Scholar 

  • Baird GS, Zacharias DA, Tsien RY (2000) Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. Proc Natl Acad Sci USA 97(22):11984–11989

    Article  CAS  Google Scholar 

  • Barkay T, Gillman M, Turner RR (1997) Effects of dissolved organic carbon and salinity on bioavailability of mercury. Appl Environ Microbiol 63(11):4267–4271

    CAS  Google Scholar 

  • Barkay T, Turner RR, Rasmussen LD, Kelly CA, Rudd JW (1998) Luminescence facilitated detection of bioavailable mercury in natural waters. Methods Mol Biol 102:231–246

    CAS  Google Scholar 

  • Belkin S (2003) Microbial whole-cell sensing systems of environmental pollutants. Curr Opin Microbiol 6(3):206–212

    Article  CAS  Google Scholar 

  • Benbelkacem H, Mathe S, Debellefontaine H (2004) Taking mass transfer limitation into account during ozonation of pollutants reacting fairly quickly. Water Sci Technol 49(4):25–30

    CAS  Google Scholar 

  • Casavant NC, Thompson D, Beattie GA, Phillips GJ, Halverson LJ (2003) Use of a site-specific recombination-based biosensor for detecting bioavailable toluene and related compounds on roots. Environ Microbiol 5(4):238–249

    Article  CAS  Google Scholar 

  • Cebron A, Louvel B, Faure P, France-Lanord C, Chen Y, Murrell JC, Leyval C (2011) Root exudates modify bacterial diversity of phenanthrene degraders in PAH-polluted soil but not phenanthrene degradation rates. Environ Microbiol 13(3):722–736

    Article  CAS  Google Scholar 

  • Close D, Xu T, Smartt A, Rogers A, Crossley R, Price S, Ripp S, Sayler G (2012) The evolution of the bacterial luciferase gene cassette (lux) as a real-time bioreporter. Sensors (Basel) 12(1):732–752

    Article  CAS  Google Scholar 

  • Deepthike HU, Tecon R, Van Kooten G, Van der Meer JR, Harms H, Wells M, Short J (2009) Unlike PAHs from Exxon Valdez crude oil, PAHs from Gulf of Alaska coals are not readily bioavailable. Environ Sci Technol 43(15):5864–5870

    Article  CAS  Google Scholar 

  • Echols KR, Brumbaugh WG, Orazio CE, May TW, Poulton BC, Peterman PH (2008) Distribution of pesticides, PAHs, PCBs, and bioavailable metals in depositional sediments of the lower Missouri River, USA. Arch Environ Contam Toxicol 55(2):161–172

    Article  CAS  Google Scholar 

  • Gao Y, Zhu L (2004) Plant uptake, accumulation and translocation of phenanthrene and pyrene in soils. Chemosphere 55(9):1169–1178

    Article  CAS  Google Scholar 

  • Hakkila K, Maksimow M, Karp M, Virta M (2002) Reporter genes lucFF, luxCDABE, gfp, and dsred have different characteristics in whole-cell bacterial sensors. Anal Biochem 301(2):235–242

    Article  CAS  Google Scholar 

  • Hansen LH, Sorensen SJ (2000) Versatile biosensor vectors for detection and quantification of mercury. FEMS Microbiol Lett 193(1):123–127

    Article  CAS  Google Scholar 

  • Impellitteri CA, Saxe JK, Cochran M, Janssen GM, Allen HE (2003) Predicting the bioavailability of copper and zinc in soils: modeling the partitioning of potentially bioavailable copper and zinc from soil solid to soil solution. Environ Toxicol Chem 22(6):1380–1386

    Article  CAS  Google Scholar 

  • Ivask A, Virta M, Kahru A (2002) Construction and use of specific luminescent recombinant bacterial sensors for the assessment of bioavailable fraction of cadmium, zinc, mercury and chromium in the soil. Soil Biol Biochem 34(10):1439–1447

    Article  CAS  Google Scholar 

  • Ivask A, Francois M, Kahru A, Dubourguier HC, Virta M, Douay F (2004) Recombinant luminescent bacterial sensors for the measurement of bioavailability of cadmium and lead in soils polluted by metal smelters. Chemosphere 55(2):147–156

    Article  CAS  Google Scholar 

  • Johnsen AR, Wick LY, Harms H (2005) Principles of microbial PAH-degradation in soil. Environ Pollut 133(1):71–84

    Article  CAS  Google Scholar 

  • Kim Y, Webster DA, Stark BC (2003) Evidence for stable transformation of Pseudomonas aeruginosa with a pUC-based plasmid. Biotechnol Lett 25(12):959–962

    Article  CAS  Google Scholar 

  • Kohler S, Belkin S, Schmid RD (2000) Reporter gene bioassays in environmental analysis. Fresenius J Anal Chem 366(6–7):769–779

    CAS  Google Scholar 

  • Kong H, He J, Gao Y, Wu H, Zhu X (2011) Cosorption of phenanthrene and mercury(II) from aqueous solution by soybean stalk-based biochar. J Agric Food Chem 59(22):12116–12123

    Article  CAS  Google Scholar 

  • Leonardi V, Giubilei MA, Federici E, Spaccapelo R, Sasek V, Novotny C, Petruccioli M, D’Annibale A (2008) Mobilizing agents enhance fungal degradation of polycyclic aromatic hydrocarbons and affect diversity of indigenous bacteria in soil. Biotechnol Bioeng 101(2):273–285

    Article  CAS  Google Scholar 

  • Lewis MA, Quarles RL, Dantin DD, Moore JC (2004) Evaluation of a Florida coastal golf complex as a local and watershed source of bioavailable contaminants. Mar Pollut Bull 48(3–4):254–262

    Article  CAS  Google Scholar 

  • MacLeod CJ, Morriss AW, Semple KT (2001) The role of microorganisms in ecological risk assessment of hydrophobic organic contaminants in soils. Adv Appl Microbiol 48:171–212

    Article  CAS  Google Scholar 

  • Martin F, Torelli S, Le Paslier D, Barbance A, Martin-Laurent F, Bru D, Geremia R, Blake G, Jouanneau Y (2012) Betaproteobacteria dominance and diversity shifts in the bacterial community of a PAH-contaminated soil exposed to phenanthrene. Environ Pollut 162:345–353

    Article  CAS  Google Scholar 

  • Marx CJ, Lidstrom ME (2001) Development of improved versatile broad-host-range vectors for use in methylotrophs and other Gram-negative bacteria. Microbiology 147(Pt 8):2065–2075

    CAS  Google Scholar 

  • Nadim A (2009) Modeling of mass transfer limitation in biomolecular assays. Ann NY Acad Sci 1161:34–43

    Article  Google Scholar 

  • Paitan Y, Biran I, Shechter N, Biran D, Rishpon J, Ron EZ (2004) Monitoring aromatic hydrocarbons by whole cell electrochemical biosensors. Anal Biochem 335(2):175–183

    Article  CAS  Google Scholar 

  • Peijnenburg W, Sneller E, Sijm D, Lijzen J, Traas T, Verbruggen E (2004) Implementation of bioavailability in standard setting and risk assessment. Environ Sci 11(3):141–149

    Google Scholar 

  • Petanen U, Romantschuk M (2002) Use of bioluminescent bacterial sensors as an alternative method for measuring heavy metals in soil extracts. Anal Chim Acta 456(1):55–61

    Article  CAS  Google Scholar 

  • Petanen T, Virta M, Karp M, Romantschuk M (2001) Construction and use of broad host range mercury and arsenite sensor plasmids in the soil bacterium Pseudomonas fluorescens OS8. Microb Ecol 41(4):360–368

    CAS  Google Scholar 

  • Rastall AC, Neziri A, Vukovic Z, Jung C, Mijovic S, Hollert H, Nikcevic S, Erdinger L (2004) The identification of readily bioavailable pollutants in Lake Shkodra/Skadar using semipermeable membrane devices (SPMDs), bioassays and chemical analysis. Environ Sci Pollut Res Int 11(4):240–253

    Article  CAS  Google Scholar 

  • Reid BJ, Stokes JD, Jones KC, Semple KT (2000) Nonexhaustive cyclodextrin-based extraction technique for the evaluation of PAH bioavailability. Environ Sci Technol 34(15):3174–3179

    Article  CAS  Google Scholar 

  • Selifonova O, Burlage R, Barkay T (1993) Bioluminescent sensors for detection of bioavailable Hg(II) in the environment. Appl Environ Microbiol 59(9):3083–3090

    CAS  Google Scholar 

  • Semple DM, Ramsden F, McIntosh AM (2003) Reduced binocular depth inversion in regular cannabis users. Pharmacol Biochem Behav 75(4):789–793

    Article  CAS  Google Scholar 

  • Shuttleworth KL, Cerniglia CE (1995) Environmental aspects of PAH biodegradation. Appl Biochem Biotechnol 54(1–3):291–302

    Article  CAS  Google Scholar 

  • Stiner L, Halverson LJ (2002) Development and characterization of a green fluorescent protein-based bacterial biosensor for bioavailable toluene and related compounds. Appl Environ Microbiol 68(4):1962–1971

    Article  CAS  Google Scholar 

  • Tauriainen S, Karp M, Chang W, Virta M (1997) Recombinant luminescent bacteria for measuring bioavailable arsenite and antimonite. Appl Environ Microbiol 63(11):4456–4461

    CAS  Google Scholar 

  • Tecon R, Wells M, van der Meer JR (2006) A new green fluorescent protein-based bacterial biosensor for analysing phenanthrene fluxes. Environ Microbiol 8(4):697–708

    Article  CAS  Google Scholar 

  • Toyooka T, Ibuki Y (2007) DNA damage induced by coexposure to PAHs and light. Environ Toxicol Pharmacol 23(2):256–263

    Article  CAS  Google Scholar 

  • Tusseau-Vuillemin MH, Gourlay C, Lorgeoux C, Mouchel JM, Buzier R, Gilbin R, Seidel JL, Elbaz-Poulichet F (2007) Dissolved and bioavailable contaminants in the Seine river basin. Sci Total Environ 375(1–3):244–256

    Article  CAS  Google Scholar 

  • Van der Meer JR, Tropel D, Jaspers M (2004) Illuminating the detection chain of bacterial bioreporters. Environ Microbiol 6(10):1005–1020

    Article  CAS  Google Scholar 

  • Vrana B, Paschke A, Popp P, Schuurmann G (2001) Use of semipermeable membrane devices (SPMDs). Determination of bioavailable, organic, waterborne contaminants in the industrial region of Bitterfeld, Saxony-Anhalt, Germany. Environ Sci Pollut Res Int 8(1):27–34

    Article  CAS  Google Scholar 

  • Wang R, Liu G, Chou CL, Liu J, Zhang J (2010) Environmental assessment of PAHs in soils around the Anhui Coal District, China. Arch Environ Contam Toxicol 59(1):62–70

    Article  CAS  Google Scholar 

  • Wei H, Cheng H, Ting M, Wen-Hui Z, Xian-Gui L (2010) A chromosomally based luminescent bioassay for mercury detection in red soil of China. Appl Microbiol Biotechnol 87(3):981–989

    Article  CAS  Google Scholar 

  • Willardson BM, Wilkins JF, Rand TA, Schupp JM, Hill KK, Keim P, Jackson PJ (1998) Development and testing of a bacterial biosensor for toluene-based environmental contaminants. Appl Environ Microbiol 64(3):1006–1012

    CAS  Google Scholar 

  • Yan J, Wang L, Fu PP, Yu H (2004) Photomutagenicity of 16 polycyclic aromatic hydrocarbons from the US EPA priority pollutant list. Mutat Res 557(1):99–108

    Article  CAS  Google Scholar 

  • Yang L, Wang Z, Xu L (2006) Simultaneous determination of phenols (bibenzyl, phenanthrene, and fluorenone) in Dendrobium species by high-performance liquid chromatography with diode array detection. J Chromatogr A 1104(1–2):230–237

    Article  CAS  Google Scholar 

  • Zhong WH, Cai ZC, Zhang H (2007) Effects of long-term application of inorganic fertilizers on biochemical properties of a rice-planting red soil. Pedosphere 17(4):419–428

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The project was financially supported by the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (Grant No. 10KJD610001) and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions. The authors thank the anonymous reviewers for their valuable comments and suggestions that greatly improved this manuscript.

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Correspondence to D. Chuan-Chao.

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Wei, H., Ze-Ling, S., Le-Le, C. et al. Specific detection of bioavailable phenanthrene and mercury by bacterium reporters in the red soil. Int. J. Environ. Sci. Technol. 11, 685–694 (2014). https://doi.org/10.1007/s13762-013-0216-1

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  • DOI: https://doi.org/10.1007/s13762-013-0216-1

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