Abstract
Investigation of bacterial chromate tolerance has mostly focused on strains originating from polluted sites. In the present study, we isolated 33 chromate tolerant strains from diverse environments harbouring varying concentrations of chromium (Cr). All of these strains were able to grow on minimal media with at least 2 mM hexavalent chromium (Cr(VI)) and their classification revealed that they belonged to 12 different species and 8 genera, with a majority (n = 20) being affiliated to the Bacillus cereus group. Selected B. cereus group strains were further characterised for their chromate tolerance level and the ability to remove toxic Cr(VI) from solution. A similar level of chromate tolerance was observed in isolates originating from environments harbouring high or low Cr. Reference B. cereus strains exhibited the same Cr(VI) tolerance which indicates that a high chromate tolerance could be an intrinsic group characteristic. Cr(VI) removal varied from 22.9% (strain PCr2a) to 98.5% (strain NCr4). Strains NCr1a and PCr12 exhibited the ability to grow to the greatest extent in Cr(VI) containing media (maximum growth of 65.3% and 64.9% relative to that in the absence of Cr(VI), respectively) accompanied with high chromate removal activity (73.7% and 74.4%, respectively), making them prime candidates for the investigation of chromate tolerance mechanisms in Gram-positive bacteria and Cr(VI) bioremediation applications.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ackerley DF, Gonzalez CF, Park CH et al (2004) Chromate-reducing properties of soluble flavoproteins from Pseudomonas putida and Escherichia coli. Appl Environ Microbiol 70:873–882
Ackerley DF, Barak Y, Lynch SV et al (2006) Effect of chromate stress on Escherichia coli K-12. J Bacteriol 188:3371–3381. https://doi.org/10.1128/JB.188.9.3371-3381.2006
Ahemad M (2014) Bacterial mechanisms for Cr(VI) resistance and reduction: an overview and recent advances. Folia Microbiol (Praha) 59:321–332. https://doi.org/10.1007/s12223-014-0304-8
Alam MZ, Ahmad S, Malik A (2011) Prevalence of heavy metal resistance in bacteria isolated from tannery effluents and affected soil. Environ Monit Assess 178:281–291. https://doi.org/10.1007/s10661-010-1689-8
Alvarez AH, Moreno-Sánchez R, Cervantes C (1999) Chromate efflux by means of the ChrA chromate resistance protein from Pseudomonas aeruginosa. J Bacteriol 181:7398–7400
Atlas R (2010) Handbook of microbiological media, 4th edn. CRC Press, Boca Raton
Banerjee S, Joshi SR, Mandal T, Halder G (2017) Insight into Cr 6 + reduction efficiency of Rhodococcus erythropolis isolated from coalmine waste water. Chemosphere 167:269–281. https://doi.org/10.1016/j.chemosphere.2016.10.012
Barak Y, Ackerley DF, Dodge CJ et al (2006) Analysis of novel soluble chromate and uranyl reductases and generation of an improved enzyme by directed evolution. Appl Environ Microbiol 72:7074–7082. https://doi.org/10.1128/AEM.01334-06
Bencheikh-Latmani R, Obraztsova A, Mackey MR et al (2007) Toxicity of Cr(III) to Shewanella sp. strain MR-4 during Cr(VI) reduction. Environ Sci Technol 41:214–220. https://doi.org/10.1021/es0622655
Bharagava RN, Mishra S (2018) Hexavalent chromium reduction potential of Cellulosimicrobium sp. isolated from common effluent treatment plant of tannery industries. Ecotoxicol Environ Saf 147:102–109. https://doi.org/10.1016/j.ecoenv.2017.08.040
Blackwood KS, Turenne CY, Harmsen D et al (2004) Reassessment of sequence-based targets for identification of bacillus species. J Clin Microbiol 42:1626–1630. https://doi.org/10.1128/JCM.42.4.1626
Branco R, Morais PV (2013) Identification and characterization of the transcriptional regulator chrb in the chromate resistance determinant of ochrobactrum tritici 5bvl1. PLoS ONE 8:e77987. https://doi.org/10.1371/journal.pone.0077987
Branco R, Alpoim MC, Morais PV (2004) Ochrobactrum tritici strain 5bvl1—characterization of a Cr(VI)-resistant and Cr(VI)-reducing strain. Can J Microbiol 50:697–703. https://doi.org/10.1139/w04-048
Branco R, Chung A-P, Veríssimo A, Morais PV (2005) Impact of chromium-contaminated wastewaters on the microbial community of a river. FEMS Microbiol Ecol 54:35–46. https://doi.org/10.1016/j.femsec.2005.02.014
Branco R, Chung AP, Johnston T et al (2008) The chromate-inducible chrBACF operon from the transposable element TnOtChr confers resistance to chromium(VI) and superoxide. J Bacteriol 190:6996–7003. https://doi.org/10.1128/JB.00289-08
Branco R, Cristóvão A, Morais PV (2013) Highly sensitive, highly specific whole-cell bioreporters for the detection of chromate in environmental samples. PLoS ONE 8:e54005. https://doi.org/10.1371/journal.pone.0054005
BSI group (2012) BS EN 15934:2012. Sludge, treated biowaste, soil and waste—calculation of dry matter fraction after determination of dry residue or water content
Caliz J, Montserrat G, Martí E et al (2012) The exposition of a calcareous Mediterranean soil to toxic concentrations of Cr, Cd and Pb produces changes in the microbiota mainly related to differential metal bioavailability. Chemosphere 89:494–504. https://doi.org/10.1016/j.chemosphere.2012.05.002
Camargo FAO, Okeke BC, Bento FM, Frankenberger WT (2005) Diversity of chromium-resistant bacteria isolated from soils contaminated with dichromate. Appl Soil Ecol 29:193–202. https://doi.org/10.1016/j.apsoil.2004.10.006
Chatterjee S, Sau GB, Mukherjee SK (2009) Plant growth promotion by a hexavalent chromium reducing bacterial strain, Cellulosimicrobium cellulans KUCr3. World J Microbiol Biotechnol 25:1829–1836. https://doi.org/10.1007/s11274-009-0084-5
Chen Z, Huang Z, Cheng Y et al (2012) Cr(VI) uptake mechanism of Bacillus cereus. Chemosphere 87:211–216. https://doi.org/10.1016/j.chemosphere.2011.12.050
Cheung KH, Gu J-D (2005) Chromate reduction by Bacillus megaterium TKW3 isolated from marine sediments. World J Microbiol Biotechnol 21:213–219. https://doi.org/10.1007/s11274-004-3619-9
Cheung KHH, Gu J-DD (2007) Mechanism of hexavalent chromium detoxification by microorganisms and bioremediation application potential: a review. Int Biodeterior Biodegrad 59:8–15. https://doi.org/10.1016/j.ibiod.2006.05.002
Cheung K, Lai H, Gu J (2006) Membrane-associated hexavalent chromium reductase of Bacillus megaterium TKW3 with induced expression. J Microbiol Biotechnol 16:855
Chirwa E, Molokwane P (2011) Biological Cr(VI) reduction: microbial diversity, kinetics and biotechnological solutions to pollution. In: Sofo A (ed) Biodiversity. InTech, London, pp 75–100
Chowdhury M, Mostafa MG, Biswas TK et al (2015) Characterization of the effluents from leather processing industries. Environ Process 2:173–187. https://doi.org/10.1007/s40710-015-0065-7
Claude KZG, Adolphe Z (2013) Proteomic analysis of chromate response in Staphylococcus saprophyticus isolated from a fly ash dumping site. Afr J Biotechnol 12:2322–2330. https://doi.org/10.5897/AJB12.127
Córdoba A, Vargas P, Dussan J (2008) Chromate reduction by Arthrobacter CR47 in biofilm packed bed reactors. J Hazard Mater 151:274–279. https://doi.org/10.1016/J.JHAZMAT.2007.10.072
De Bruijn JPF, Mondaca MA (2000) Chromate reduction by Serratia marcescens immobilized on activated carbon. Toxicol Environ Chem 76:125–135. https://doi.org/10.1080/02772240009358923
Dey S, Paul AK (2015) Hexavalent chromate reduction during growth and by immobilized cells of Arthrobacter sp. SUK 1205. Sci Technol Dev 34:158–168. https://doi.org/10.3923/std.2015.158.168
Dhal B, Thatoi HN, Das NN, Pandey BD (2013) Chemical and microbial remediation of hexavalent chromium from contaminated soil and mining/metallurgical solid waste: a review. J Hazard Mater 250–251:272–291. https://doi.org/10.1016/j.jhazmat.2013.01.048
Díaz-Magaña A, Aguilar-Barajas E, Moreno-Sánchez R et al (2009) Short-chain chromate ion transporter proteins from Bacillus subtilis confer chromate resistance in Escherichia coli. J Bacteriol 191:5441–5445. https://doi.org/10.1128/JB.00625-09
Dworkin M, Falkow S, Rosenberg E et al (eds) (2006) The Prokaryotes a handbook on the biology of bacteria third edition volume 4: Bacteria: Firmicutes, Cyanobacteria, 3rd edn. Springer
Fritze D (2004) Taxonomy of the genus Bacillus and related genera: the aerobic endospore-forming bacteria. Phytopathology 94:1245–1248. https://doi.org/10.1094/PHYTO.2004.94.11.1245
Gawlik B, Bidoglio G (2006) Background values in European soils and sewage sludges. Results of a JRC-coordinated study on background values. Part III—conclusions, comments and recommendations
He M, Li X, Guo L et al (2010) Characterization and genomic analysis of chromate resistant and reducing Bacillus cereus strain SJ1. BMC Microbiol 10:221. https://doi.org/10.1186/1471-2180-10-221
He M, Li X, Liu H et al (2011) Characterization and genomic analysis of a highly chromate resistant and reducing bacterial strain Lysinibacillus fusiformis ZC1. J Hazard Mater 185:682–688. https://doi.org/10.1016/j.jhazmat.2010.09.072
He Z, Shentu J, Yang X et al (2015) Heavy metal contamination of soils: sources, indicators, and assessment. J Environ Indic 9:17–18
Henne KL, Nakatsu CH, Thompson DK, Konopka AE (2009a) High-level chromate resistance in Arthrobacter sp. strain FB24 requires previously uncharacterized accessory genes. BMC Microbiol 9:199. https://doi.org/10.1186/1471-2180-9-199
Henne KL, Turse JE, Nicora CD et al (2009b) Global proteomic analysis of the chromate response in Arthrobacter sp. Strain FB24. J Proteome Res 8:1704–1716. https://doi.org/10.1021/pr800705f
Hu P, Brodie EL, Suzuki Y et al (2005) Whole-genome transcriptional analysis of heavy metal stresses in Caulobacter crescentus. J Bacteriol 187:8437–8449. https://doi.org/10.1128/JB.187.24.8437-8449.2005
Ibrahim ASS, El-Tayeb M, Elbadawi YB, Al-Salamah AA (2011) Bioreduction of Cr (VI) by potent novel chromate resistant alkaliphilic Bacillus sp strain KSUCr5 isolated from hypersaline Soda lakes. African J Biotechnol 10:7207–7218. https://doi.org/10.5897/AJB11.1030
International Agency for Research on Cancer (2012) IARC Monographs On the evaluation of carcinogenic risks to humans volume 100c. Arsenic, Metals, Fibres And Dusts
ISO (2005) ISO method 10390:2005 soil quality—determination of pH
ISO (2009) ISO method 11277:2009 soil quality—determination of particle size distribution in mineral soil material—method by sieving and sedimentation
Jahan M, Akhtar N, Khan N et al (2015) Characterization of tannery wastewater and its treatment by aquatic macrophytes and algae. Bangladesh J Sci Ind Res 49:233–242. https://doi.org/10.3329/bjsir.v49i4.22626
Jiménez G, Urdiain M, Cifuentes A et al (2013) Description of Bacillus toyonensis sp. nov., a novel species of the Bacillus cereus group, and pairwise genome comparisons of the species of the group by means of ANI calculations. Syst Appl Microbiol 36:383–391. https://doi.org/10.1016/j.syapm.2013.04.008
Jung MY, Kim J-S, Paek WK et al (2011) Bacillus manliponensis sp. nov., a new member of the Bacillus cereus group isolated from foreshore tidal flat sediment. J Microbiol 49:1027–1032. https://doi.org/10.1007/s12275-011-1049-6
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874. https://doi.org/10.1093/molbev/msw054
Kumari V, Yadav A, Haq I et al (2016) Genotoxicity evaluation of tannery effluent treated with newly isolated hexavalent chromium reducing Bacillus cereus. J Environ Manage 183:204–211. https://doi.org/10.1016/j.jenvman.2016.08.017
Liu B, Liu G-H, Hu G-P et al (2014) Bacillus bingmayongensis sp. nov., isolated from the pit soil of Emperor Qin’s Terra-cotta warriors in China. Antonie Van Leeuwenhoek 105:501–510. https://doi.org/10.1007/s10482-013-0102-3
Liu Y, Lai Q, Göker M et al (2015) Genomic insights into the taxonomic status of the Bacillus cereus group. Sci Rep 5:14082. https://doi.org/10.1038/srep14082
Logan NA, De Vos P (2009) Genus I. Bacillus cohn 1872, 174AL. In: Vos P, De Garrity GM, Jones D et al (eds) Bergey’s manual of systematic bacteriology volume three the firmicutes, 2nd edn. Springer, Dordrecht Heidelberg, pp 21–130
Lun L, Li D, Yin Y et al (2016) Characterization of chromium waste form based on biocementation by Microbacterium sp. GM-1. Indian J Microbiol 56:353–360. https://doi.org/10.1007/s12088-016-0579-3
Mergeay M (1995) Heavy metal resistances in microbial ecosystems. In: Akkermans ADL, Van Elsas JD, De Bruijn FJ (eds) Molecular microbial ecology manual. Springer, Dordrecht, pp 439–455
Middleton SS, Latmani RB, Mackey MR et al (2003) Cometabolism of Cr(VI) by Shewanella oneidensis MR-1 produces cell-associated reduced chromium and inhibits growth. Biotechnol Bioeng 83:627–637. https://doi.org/10.1002/bit.10725
Morais PV, Branco R, Francisco R (2011) Chromium resistance strategies and toxicity: what makes Ochrobactrum tritici 5bvl1 a strain highly resistant. Biometals 24:401–410. https://doi.org/10.1007/s10534-011-9446-1
Murugavelh S, Mohanty K (2013) Isolation, identification and characterization of Cr(VI) reducing Bacillus cereus from chromium contaminated soil. Chem Eng J 230:1–9. https://doi.org/10.1016/j.cej.2013.06.049
Nancharaiah YV, Dodge C, Venugopalan VP et al (2010) Immobilization of Cr(VI) and its reduction to Cr(III) phosphate by granular biofilms comprising a mixture of microbes. Appl Environ Microbiol 76:2433–2438. https://doi.org/10.1128/AEM.02792-09
Nicholson WL, Munakata N, Horneck G et al (2000) Resistance of bacillus endospores to extreme terrestrial and extraterrestrial environments. Microbiol Mol Biol Rev 64:548–572. https://doi.org/10.1128/MMBR.64.3.548-572.2000
Nies A, Nies DH, Silver S (1989) Cloning and expression of plasmid genes encoding resistances to chromate and cobalt in Alcaligenes eutrophus. J Bacteriol 171:5065–5070
Patra RC, Malik S, Beer M et al (2010) Molecular characterization of chromium (VI) reducing potential in Gram positive bacteria isolated from contaminated sites. Soil Biol Biochem 42:1857–1863. https://doi.org/10.1016/j.soilbio.2010.07.005
Pattanapipitpaisal P, Brown NL, Macaskie LE (2001a) Chromate reduction by Microbacterium liquefaciens immobilised in polyvinyl alcohol. Biotechnol Lett 23:61–65. https://doi.org/10.1023/A:1026750810580
Pattanapipitpaisal P, Brown NL, Macaskie LE (2001b) Chromate reduction and 16S rRNA identification of bacteria isolated from a Cr(VI)-contaminated site. Appl Microbiol Biotechnol 57:257–261. https://doi.org/10.1007/s002530100758
Ramírez-Díaz MI, Díaz-Pérez C, Vargas E et al (2008) Mechanisms of bacterial resistance to chromium compounds. Biometals 21:321–332. https://doi.org/10.1007/s10534-007-9121-8
Ramisse V, Patra G, Garrigue H et al (1996) Identification and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA. FEMS Microbiol Lett 145:9–16. https://doi.org/10.1016/0378-1097(96)00358-8
Ross DS, Sjogren RE, Bartlett RJ (1981) Behavior of chromium in soils: IV. Toxicity to microorganisms1. J Environ Qual 10:145. https://doi.org/10.2134/jeq1981.00472425001000020004x
Roychowdhury R, Mukherjee P, Roy M (2016) Identification of chromium resistant bacteria from dry fly ash sample of mejia MTPS thermal power plant, West Bengal, India. Bull Environ Contam Toxicol 96:210–216. https://doi.org/10.1007/s00128-015-1692-4
Sagar S, Dwivedi A, Yadav S et al (2012) Hexavalent chromium reduction and plant growth promotion by Staphylococcus arlettae Strain Cr11. Chemosphere 86:847–852. https://doi.org/10.1016/J.CHEMOSPHERE.2011.11.031
Sambrook J, Russell DW (2001) Commonly used techniques in molecular cloning. In: Molecular cloning, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Samuel J, Paul ML, Ravishankar H et al (2013) The differential stress response of adapted chromite mine isolates Bacillus subtilis and Escherichia coli and its impact on bioremediation potential. Biodegradation 24:829–842. https://doi.org/10.1007/s10532-013-9631-8
Shakoori a R, Makhdoom M, Haq RU (2000) Hexavalent chromium reduction by a dichromate-resistant gram-positive bacterium isolated from effluents of tanneries. Appl Microbiol Biotechnol 53:348–351. https://doi.org/10.1007/s002530050033
Singh AL, Murali Krishna P, Nageswara Rao T, Sarma PN (2006) Uptake of Cr(VI) in the presence of sulphate by Bacillus mycoides in aerobic environment. Indian J Biotechnol 5:506–509
Singh A, Vyas D, Malaviya P (2016) Two-stage phyto-microremediation of tannery effluent by Spirodela polyrrhiza (L.) Schleid. and chromium resistant bacteria. Bioresour Technol 216:883–893. https://doi.org/10.1016/J.BIORTECH.2016.06.025
Thatoi H, Das S, Mishra J et al (2014) Bacterial chromate reductase, a potential enzyme for bioremediation of hexavalent chromium: A review. J Environ Manage 146:383–399. https://doi.org/10.1016/j.jenvman.2014.07.014
Torsvik V, Frida A, Daae L (1990) High diversity in DNA of soil bacteria. Appl Environ Microbiol 56:782–787
Trivedi P, Pandey A, Sa T (2007) Chromate reducing and plant growth promoting activies of psychrotrophic Rhodococcus erythropolis MtCC 7905. J Basic Microbiol 47:513–517. https://doi.org/10.1002/jobm.200700224
Turpeinen R, Kairesalo T, Häggblom MM (2004) Microbial community structure and activity in arsenic-, chromium- and copper-contaminated soils. FEMS Microbiol Ecol 47:39–50. https://doi.org/10.1016/S0168-6496(03)00232-0
Urone PF (1955) Stability of colorimetric reagent for chromium, s-diphenylcarbazide, in various solvents. Anal Chem 27:1354–1355. https://doi.org/10.1021/ac60104a048
USEPA (2007) US EPA method 7000B: Flame Atomic Absorption Spectrophotometry
Vatsouria A, Vainshtein M, Kuschk P et al (2005) Anaerobic co-reduction of chromate and nitrate by bacterial cultures of Staphylococcus epidermidis L-02. J Ind Microbiol Biotechnol 32:409–414. https://doi.org/10.1007/s10295-005-0020-0
Viti C, Giovannetti L (2001) The impact of chromium contamination on soil heterotrophic and photosynthetic microorganisms. Ann Microbiol 51:201–213
Viti C, Mini A, Ranalli G et al (2006) Response of microbial communities to different doses of chromate in soil microcosms. Appl Soil Ecol 34:125–139. https://doi.org/10.1016/j.apsoil.2006.03.003
Wang S-S, Ye S-L, Han Y-H et al (2016) Biosorption and bioaccumulation of chromate from aqueous solution by a newly isolated Bacillus mycoides strain 200AsB1. RSC Adv 6:101153–101161. https://doi.org/10.1039/C6RA23879C
Wani R, Kodam KM, Gawai KR, Dhakephalkar PK (2007) Chromate reduction by Burkholderia cepacia MCMB-821, isolated from the pristine habitat of alkaline crater lake. Appl Microbiol Biotechnol 75:627–632. https://doi.org/10.1007/s00253-007-0862-7
Zhang K, Li F (2011) Isolation and characterization of a chromium-resistant bacterium Serratia sp. Cr-10 from a chromate-contaminated site. Appl Microbiol Biotechnol 90:1163–1169. https://doi.org/10.1007/s00253-011-3120-y
Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214. https://doi.org/10.1089/10665270050081478
Zhang H, Ma Y, Liu P, Li X (2016) Multidrug resistance operon emrAB contributes for chromate and ampicillin co-resistance in a Staphylococcus strain isolated from refinery polluted river bank. Springerplus 5:1648. https://doi.org/10.1186/s40064-016-3253-7
Zheng Z, Li Y, Zhang X et al (2015) A Bacillus subtilis strain can reduce hexavalent chromium to trivalent and an nfrA gene is involved. Int Biodeterior Biodegrad 97:90–96. https://doi.org/10.1016/j.ibiod.2014.10.017
Acknowledgements
Authors would like to thank Albert Breier and Petra Olejnikova from STU Bratislava for MALDI TOF analysis. The project was financed by the SCOPES programme of the Swiss National Science Foundation (SNSF) and the Swiss Agency for Development and Cooperation (SDC) (Grant Number SCOPES IZ73Z0_152527/1) to RBL, DR and IB, and by grant APVV-14-0181 to IB and by grant from Slovak Research Agency (ITMS 26230120006).
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Tamindžija, D., Chromikova, Z., Spaić, A. et al. Chromate tolerance and removal of bacterial strains isolated from uncontaminated and chromium-polluted environments. World J Microbiol Biotechnol 35, 56 (2019). https://doi.org/10.1007/s11274-019-2638-5
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-019-2638-5