Heavy Metal and Antibiotic Resistant Bacteria Isolated from Soil Contaminated by the Sugar Industry Effluent

Paper Details

Research Paper 01/02/2016
Views (398) Download (21)

Heavy Metal and Antibiotic Resistant Bacteria Isolated from Soil Contaminated by the Sugar Industry Effluent

Saurabh Saini, Shailja Pant
Int. J. Biosci.8( 2), 36-46, February 2016.
Certificate: IJB 2016 [Generate Certificate]


In this study, antibiotic and heavy metal resistant bacteria were isolated from the agriculture field’s soil around sugar industry situated in Roorkee, District Haridwar, Uttarakhand (India).The antibiotic and heavy metal resistance profiles of the isolates were determined. Fifteen antibiotics and five heavy metals were used for reference. Four promising isolates were recovered, two gram positive E-2 and E-10 and two gram negative isolate E-7 and E-9. Detailed morphological, biochemical, and molecular characterization was done to identify the bacteria. The isolates E-2 and E-10 were identified as Bacillus licheniformis sp, and Teribacillus aidingensis sp. while E-7 and E-9 were identified as Chryseobacterium indologenes sp., and Enterobacter cloacae sp. respectively. These isolates were found resistant to heavy metals such as Ni2+, Pb2+, Cu2+, Zn2+ and Hg2+ at different concentrations ranging from 0.05 mM to 12 mM. The trend for tolerance of heavy metals by isolates was as follows: E-2:- Ni = Cu >Pb> Zn > Hg, E-7: Ni > Cu = Zn >Pb> Hg, E-9:- Pb = Ni > Cu = Zn > Hg and E-10:- Zn > Ni > Cu >Pb> Hg. The Anti-biogram pattern indicates that all isolates were showing resistance against more than five antibiotics and all are designated as multi-drugs resistant bacteria. These bacteria can be explored for bio-absorption of heavy metals from contaminated sites.


Ahemad M. 2012. Implication of Bacterial Resistance against Heavy Metals in Bioremediation: A review. The IIOAB Journal (3)3, 39-46.

Austin CB, Wright MS, Stepanauskas R, Mcarthur JV. 2006. Co-selection of antibiotic and metal resistance. TRENDS in Microbiology 14(4), 176-182.

Bauer AW, Kirby WM, Sherris JC,Turck M. 1996.Antibiotic susceptibility testing by standard single disk method. Am J Clin pathol 45, 493-496.

Bhattacherjee JW, Pathak SP, Gaur A. 1988. Antibiotic and metal tolerance of coli form bacteria isolated from Gomati river water at Luknow city. Journal of general Applied Microbiology 34, 391-399.

Chopra A K, Pathak C, Prasad G. 2009.Scenario of heavy metal contamination in agricultural soil and its management. Journal of Applied and Natural Science 1,99-108.

Claus D, Berkeley RCW. 1968. Genus pseudomonas. In Bergey’s Manual of Systematic Bacteriology. Editted by Sneath PHA, et al. Baltimore,MD: Williams and Willkins Co 2(2), 140-219.

Deeb BE. 2009. Plasmid Mediated Tolerance and Removal of Heavy Metals by Enterobacter sp. American Journal of Biochemistry and Biotechnology 5(1), 47-53.

Frank, JA, Reich C, Sharma S, Weisbaum JS, Wilson BA, Olsen GJ. 2008. Critical Evaluation of Two Primers Commonly Used for Amplification of Bacterial 16S rRNA Genes. Applied and Environmental Microbiology 74(8), 2461–2470. http://dx.doi.org/ 10.1128/AEM.02272-07

Gupta MK, Kumara K, Shrivastava A, Gauri S.2014. Bioremediation of Heavy metal polluted environment using resistant bacteria. Journal of Environmental Research and Development 8(4), 883-889.

Hookoom M, Puchooa D. 2013.Isolation and Identification of Heavy Metals Tolerant bacteria from Industrial and Agricultural areas in Mauritus. Current Research in Microbiology & Biotechnology 1, 119-123.

Ince-Yilmaz E. 2003. Metal tolerance and biosorption capacity of Bacillus circulans strain EB1. Research in Microbiology 154, 409–415.

Kirby JT,Sader SH, Walsh TR, Jones RN. 2004. Antimicrobial Susceptibility and Epidemiology of a Worldwide Collection of Chryseobacteriumspp.: Report from the SENTRY Antimicrobial Surveillance Program (1997–2001). Journal of Clinical Microbiology 42(1), 445–448.

Kumar V, Chopra AK. 2010. Influence of sugar mill effluent on physico-chemical characteristics of soil at Haridwar (Uttarakhand). India.Journal of Applied and Natural Science 2(2), 269-279.

Kocsis B, Szabó D. 2013. Antibiotic resistance mechanisms in Enterobacteriaceae. Microbial pathogens and strategies for combating them: science, technology and education (A. Méndez-Vilas, Ed.), 251-257.

Mak YM, Ho KK. 1991. An improved method for the isolation of chromosomal DNA from various bacteria and cyanobacteria. Nucleic Acids Research 20, 4101-4102.

Matyar F. 2012. Antibiotic and Heavy Metal Resistant in Bacterial Isolated from the Eastern Mediterranean Sea Coast. Bull Environ Contam Toxicol 89, 551-556. http://dx.doi.org/10.1007/s00128-012-0726-4

Monachese M, Burton JP, Reid G. 2012.Bioremediation and Tolerance of humans to heavy Metals through Microbial process: a Potential Role for Probiotics? Applied and environmental Microbiology, 78(18), 6397-6404.

Nanda M, Sharma D, Kumar A. 2011. Removal of Heavy Metals from Industrial Effluent Using Bacteria. International Journal of Environment Sciences 2(2), 781-787.

Nies DH. 1999. Microbial heavy-metal resistance. Appl Microbial Biotechnol 51, 730-750. http://dx.doi.org/10.1007/s002530051457

Pontes DS, Pinheriro FA,  Lima CI, Guedes RLM, Cursino L, Barbosa F, Santos FR, Chartone SE, Nascimento AMA. 2009. Multiple antimicrobial resistances of gram negative bacteria from natural ologotrophic lakes under distinct antropogenic influence in a tropical region. Microb Ecol 58, 762-772 http://dx.doi.org/10.1007/s00248-009-9539-3

Raja CE, Anbazhagan K, Selvam GS. 2006. Isolation and characterization of a metal-resistant Pseudomonas aeruginosa strain. World Journal of Microbiology & Biotechnology 22, 577-585. http://dx.doi.org/10.1007/s11274-005-9074-4

Rajbanshi A. 2008. Study on Heavy Metal Resistant Bacteria in Guheswori Sewage Treatment Plant. Our Nature 6, 52-57. http://dx.doi.org/10.3126/on.v6i1.1655

Rajendran P, Muthukrishnan, J, Gunasekaran P. 2003.Microbes in heavy metal remediation. Indian Journal of Experimental Biology 41, 935-944.

Rajkumar B, Sharma GD, Paul AK. 2012. Isolation and Characterization of Heavy Metal Resistant Bacteria from Barak River Contaminated with Pulp Paper Mill Effluent, South Assam. Bull Environ Contam Toxicol 89, 263–268.

Rani JM, Hemambika B, Hemapriya J, Rajeshkannan V. 2010. Comparative Assessment of Heavy Metal Removal by Immobilized and Dead Bacterial Cells: A Biosorption Approach. Global Journal of Environment Research 4(1), 23-30.

Y Ren, Ren Y, Zhou Z, Guo X, Li Y, Feng L, Wang L. 2010.Complete Genome Sequence of Enterobacter  cloacae  subsp. Cloacae  Type Strain ATCC 13047. Journal of Bacteriology 192(9), 2463–2464. http://dx.doi.org/10.1128/JB.00067-10

Samanta A, Bera P, Khatun M, Sinha C, Pal P, Lalee A, Mandal A. 2012. An investigation on heavy metal tolerance and antibiotic resistance properties of bacterial strain Bacillus sp. isolated from municipal waste. Journal Microbiolology Biotechnology Research2 (1), 178-189.

Saranraj P, Stella D. 2012.Bioremediation of sugar mill effluent by Immobilized Bacterial Consortium. International Journal of Research in Pure and Applied Microbiology 2(4), 43-48.

Saranraj P, Stella D. 2014.Impactof Sugar Mill Effluent to Environment and Bioremediation.A Review.World Applied Sciences Journal 30(3), 299-316.

Silver S. 1996.Bacterial resistance to toxic metal ions-a review.Gene, 179 (1), 9-19. http://dx.doi.org/10.1016/S0378-1119(96)00323-X

Silver S, Mishra TK . 1988. Plasmid-mediated heavy metal resistance. Annual Review of Microbiology 42,717-737. http://dx.doi.org/10.1146/annurev.micro.42.1.717

Sneath PHA, Mair NS, Sharpe ME, Holt JG. 1986. Bergey’s Manual of Sytematic Bacteriology 2, 140-219,

Stiles ME, NG Lai-King. 1981. Biochemical Characteristics and Identification of Enterobacteriaceae Isolated from Meats. Appl. and Environ. Micro 41(3), 639-645,

Suriya J, Bharathiraja S, Rajasekaran R. 2013. Biosorption of Heavy Metals by Biomass of Enterobacter Cloacae Isolated From Metal-Polluted Soils. International Journal of ChemTechResearch 5(3),1329-1338.

Vandamme P, Bernardet JF, Segers P, Kersters K, Holmes B. 1994. New Perspectives in the Classification of the Flavobacteria: Description of Chryseo-bacteriumgen. nov., Bergeyellagen. nov., and Empedobacternorn.rev. International Journal of Systematic Bacteriology 44(4), 827-831.

Veith B, Herzberg C, Steckel S, Feesche J, Maurer KH, Ehrenreich P, Bäumer S, Henne A, Liesegang H, Merkl R, Ehrenreich A, Gottschalk G. 2004.The Complete Genome Sequence of Bacillus licheniformisDSM13, an Organism with Great Industrial Potential. Journal of Molecular Microbiology Biotechnology 7, 204–211.

Verma T, Srinath T, Gadpayle RU, Ramtake PW, Hans RK, Garg SK. 2001. Chromate tolerant bacteria isolated from tannery effluent. Bioresource Technology 78, 31-35. http://dx.doi.org/10.1016/S0960-8524(00)00168-1

Wu G, Kang H, Zhang X, Shao H, Chu L, Ruan C.  2010.A  critical  review  on  the  bio-removal  of hazardous  heavy  metals  from  contaminated  soils: Issues, progress, eco-environmental concerns and opportunities. Journal of Hazardous Materials 174, 1-8. http://dx.doi.org/10.1016/j.jhazmat.2009.09.113

Zeba B, Simporé PJ, Nacoulma OG, Frèrej M. 2005. Identification of metallo-b-lactamase from a clinical isolate at Saint Camillle medical Center of Ouagadougou, Burkina Faso. African Journal of Biotechnology 4(3), 286-288.

Zeba B, Luca FD, Dubus A, Delmarcelle M, Simpore J, Nacoulma OG, Rossolini GM, Fre`Re JM, Docquier JD. 2009. IND-6, a Highly Divergent IND-Type Metallo-Lactamase from Chryseobacterium indologenes Strain 597 Isolated in Burkina Faso. Antimicrobial agents and Chemotherapy, 53(10), 4320-4326. http://dx.doi.org /10.1128/AAC.01607-08