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PHYTOREMEDIATION POTENTIAL OF VETIVER GRASS [CHRYSOPOGON ZIZANIOIDES (L.)] FOR TETRACYCLINE a
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Rupali Datta , Padmini Das , Stephanie Smith , Pravin Punamiya b
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, Dil M. Ramanathan , Ramana Reddy & Dibyendu Sarkar
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Department of Biological Sciences, Michigan Technological University, Houghton, Michigan, USA b
Department of Earth and Environmental Studies, Montclair State University, Upper Montclair, New Jersey, USA c
New Jersey Center for Science, Technology and Mathematics, Kean University, Union, New Jersey, USA Accepted author version posted online: 05 Jul 2012.
To cite this article: Rupali Datta, Padmini Das, Stephanie Smith, Pravin Punamiya, Dil M. Ramanathan, Ramana Reddy & Dibyendu Sarkar (2013): PHYTOREMEDIATION POTENTIAL OF VETIVER GRASS [CHRYSOPOGON ZIZANIOIDES (L.)] FOR TETRACYCLINE, International Journal of Phytoremediation, 15:4, 343-351 To link to this article: http://dx.doi.org/10.1080/15226514.2012.702803
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International Journal of Phytoremediation, 15:343–351, 2013 C Taylor & Francis Group, LLC Copyright ISSN: 1522-6514 print / 1549-7879 online DOI: 10.1080/15226514.2012.702803
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PHYTOREMEDIATION POTENTIAL OF VETIVER GRASS [CHRYSOPOGON ZIZANIOIDES (L.)] FOR TETRACYCLINE Rupali Datta,1 Padmini Das,2 Stephanie Smith,1 Pravin Punamiya,2 Dil M. Ramanathan,3 Ramana Reddy,1 and Dibyendu Sarkar2 1
Department of Biological Sciences, Michigan Technological University, Houghton, Michigan, USA 2 Department of Earth and Environmental Studies, Montclair State University, Upper Montclair, New Jersey, USA 3 New Jersey Center for Science, Technology and Mathematics, Kean University, Union, New Jersey, USA The presence of veterinary and human antibiotics in soil and surface water is an emerging environmental concern. The current study was aimed at evaluating the potential of using vetiver grass as a phytoremediation agent in removing Tetracycline (TC) from aqueous media. The study determined uptake, translocation, and transformation of TC in vetiver grass as function of initial antibiotic concentrations and exposure time. Vetiver plants were grown for 60 days in a greenhouse in TC contaminated hydroponic system. Preliminary results show that complete removal of tetracycline occurred within 40 days in all TC treatments. Initial concentrations of TC had significant effect (p < 0.0001) on the kinetics of removal. Tetracycline was detected in the root as well as shoot tissues, confirming uptake and rootto-shoot translocation. Liquid-chromatography-tandem-mass-spectrometry analysis of plant tissue samples suggest presence of metabolites of TC in both root and shoot tissues of vetiver grass. The current data is encouraging and is expected to aid in developing a cost-effective, in-situ phytoremediation technique to remove TC group of antibiotics from wastewater. KEY WORDS: tetracycline, antibiotics, phytoremediation, vetiver grass
INTRODUCTION Soil and surface water contamination with human and veterinary antibiotics (VAs) is an emerging environmental concern. Four compounds namely tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC), and doxycycline (DC) belong to a class of antibiotics which are commonly called as tetracyclines (TCs). The TCs are broad-spectrum antibacterials which are widely used as human antibiotics (TC, OTC) as well as in animal feed (CTC, OTC, TC, and DC) to maintain health and improve growth efficiency (Kemper 2008). Capelton et al. (2006) prioritized 83 veterinary medicines for detailed risk assessments based on their usage, potential to reach the environment, and toxicity profile. CTC, OTC, and TC ranked 16, 28, and 31 respectively in their priority list with high usage and potential to reach the environment and priority for the risk assessment (Capelton
Address correspondence to Rupali Datta, Department of Biological Sciences, Michigan Technological University, 1400 Townsend Drive, Houghton, MI, USA. E-mail:
[email protected] 343
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et al. 2006). Primary usage of CTC is restricted to cattle and pigs, OTC is mostly used in veterinary medicines (cattle, sheep, and pigs), but also used as a human drug; whereas TC is one of the most widely used human antibiotics, that is also used in horse, sheep and pigs (Kemper 2008). In community hospitals, TC along with penicillin and ampicillin are the most used antibiotics, which account for about half of the antibiotics given to patients (Sheckler and Bennett 1970). Being bioactive substances, antibiotics are highly effective at low doses and cannot be completely eliminated by the animal or human organisms, and thus excreted after short time of retention (Kemper 2008). Excretion rates of TCs vary between 40–90% depending on the mode of application, the excreting species, and time after administration (Berger, Petersen, and Buning-Pfau 1986; Haller et al. 2001; Halling-Sørensen et al. 2001). Intracorporal degradation of the antibiotics often takes place in the feces, but after excretion these metabolites can be transformed back to their parent compounds (Langhammer 1989). In last decade, many studies from several countries found occurrences of TCs in sewage influent and effluent, in surface water, groundwater and even drinking water (Kemper 2008; Sarmah, Meyer, and Boxall 2006). For instance, presence of TCs in river water (1µg L−1) was first reported in England (Watts et al. 1982). Tetracyclines (ranged from 1000 µg L−1) were most frequently detected in the wastewater lagoons at swine and poultry animal feeding operations from six states of United States (Meyer et al. 2003). A U.S. Geological Survey study found large proportions of TCs ( 70% of the total TC in the plant tissues were located in the shoot which is in contrast with the findings of Migliore, Cozzolino, and Fiori (2003) and Kong et al. (2007) who suggested that the antibiotics mainly accumulate in the root tissues of the plants. Root to shoot translocation factors (TF) (calculated as the TC in shoot/TC in root) of TC in vetiver grass significantly (p = 0.0003) increased with increasing initial TC concentrations in solution (Table 1). The high TF (> 1) across higher TC treatments (10 and 15 mg L−1) indicates the potential of vetiver grass as an effective phytoremediation agent. Unknown peaks other than TC found in the HPLC chromatogram of all root and shoot samples led us to further analyze these samples using liquid chromatography-tandem mass spectrometry. Figure 2 shows the LC-MS/MS chromatograms of root (2A) and shoot (2B) samples. In all root samples, major peaks were found at higher m/z than that of TC (m/z = 445). In contrast, in the shoot samples, major peaks were at lower m/z as compared to the TC. This suggests presence of TC conjugates in the root samples, whereas in shoot, the degradation products of tetracycline prevail. Absence of these peaks in the hydroponic solution and in the control plants not treated with TC suggests that these unknown peaks are plant metabolites of TC. Calculations were also performed to check for the presence of solvents conjugate, mainly ACN-TC and MeOH-TC and salt conjugate, Na-TC and K-TC. Analysis by mass spec revealed absence of both the solvent and salt conjugate with TC. Ongoing studies involve performing liquid chromatography–tandem mass spectrometry to identify the newly formed daughter compounds and metabolites via Meta ID (metabolite identification) in the plant tissue. LTQ-Orbitrap will be used to determine and identify and plant conjugate /non-conjugate metabolites using ion trap (both positive and negative mode simultaneously). Future studies aims towards developing solid phase extraction (SPE) techniques for extraction of compounds from plant tissue and robust and sensitive mass spectrometry method to identify and quantify TC and its metabolites. Our ultimate goal is to develop an in situ biological treatment system to remove TCs from manure stockpiles at lagoons and wastewater using constructed wetlands. This study is the first step towards our long-term goal of remediating TCs contaminated soil and water
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Figure 2 LC-MS-MS chromatograms of the vetiver root (2A) and shoot (2B) suggesting the presence of TC metabolites.
using vetiver grass. Results demonstrated the ability of vetiver grass to uptake, translocate, and biotransform tetracycline. Given the huge biomass of vetiver grass compared to any similar species and it’s tolerance of a wide variety of environmental conditions and matrices indicate its potential as a phytoremediation agent in aqueous systems. The effectiveness of vetiver grass in removing organic contaminants has been reported in various previous reports (Makris et al. 2007; Das et al 2010; Marcacci et al. 2006). Tetracycline from the aqueous media and structural similarity between tetracycline group of antibiotics led us speculate that vetiver grass would also be able to remediate OTC and CTC. We are in process of designing a greenhouse experiment to study the phytoremediation potential (uptake and metabolism of detoxification) of vetiver grass in remediating TC, OTC, and CTC from contaminated municipal wastewater and manure lagoons.
ABBREVIATIONS TC OTC CTC HPLC LC-MS/MS
Tetracycline Oxytetracycline Chlortetracycline High performance liquid chromatography Liquid chromatography tandem mass spectrometry.
ACKNOWLEDGMENTS Student author SS acknowledges the Summer Undergraduate Research Fellowship provided by Michigan Technological University. Student authors PD, PP and RR acknowledge their respective universities for Graduate Assistantships.
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REFERENCES Andra SS, Datta R, Sarkar D, Makris KC, Mullens CP, Sahi SV, Bach SBH. 2009. Induction of Lead-Binding Phytochelatins in Vetiver Grass [Vetiveria zizanioides (L.)]. J Environ Qual 38:868–877. Berger K, Petersen B, Buning-Pfaue H. 1986. Persistence of drugs occurring in liquid manure in the food chain. Archiv Lebensmittelhygiene 37:99–102. Boonsaner M, Hawker DW. 2010. Accumulation of oxytetracycline and norfloxacin from saline soil by soybeans. Sci Total Environ 408:1731–1737. Bradel BG, Preil W, Jeske H. 2000. Remission of the free-branching pattern of Euphorbia pulcherrima by tetracycline treatment. J Phytopathol 148:587–590. Campagnolo ER, Johnson KR, Karpati A, Rubin CS, Kolpin DW Meyer MT, Esteban JE, Currier RW, Smith K, Thug KM, McGeehin M. 2002. Antimicrobial residues in animal waste and water resources proximal to large-scale swine and poultry feeding operations. Sci Total Environ 299:89–95. Capleton AC, Courage C, Rumsby P, Holmes P, Stutt E, Boxall ABA, Levy LS. 2006. Prioritising veterinary medicines according to their potential indirect human exposure and toxicity profile. Toxicol Lett 163:213–223. Danh LT, Truong P, Mammucari R, Tran T, Foster N. 2009. A Choice Plant for Phytoremediation of Heavy Metals and Organic Wastes Intern. J Phytorem 11:664–691. Dalton PA, Smith RJ, Truong PNV. 1996. Vetiver grass hedges for erosion control on a cropped flood plain: Hedge hydraulics. Agric Water Manag 31:91–104. Das P, Datta R, Makris KC, Sarkar D. 2010. Vetiver grass is capable of removing TNT from soil in the presence of urea. Environ Pollut 158:1980–1983. Datta R, Quispe M, Sarkar D. 2011. Greenhouse Study on the Phytoremediation Potential of Vetiver Grass, Chrysopogon zizanioides L., in Arsenic-Contaminated Soils. Bulletin Environ Contamin Toxicol 86:124–128. Dolliver H, Kumar K, Gupta S, Singh A. 2008. Application of Enzyme-Linked Immunosorbent Assay Analysis for Determination of Monensin in Environmental Samples. J Environ Qual 37:1220–1226. Farkas MH, Berry JO, Aga DS. 2007. Chlortetracycline Detoxification in Maize via Induction of Glutathione S-Transferases after Antibiotic Exposure. Environ Sci Technol 41:1450–1456. Fritz JW, Zuo Y. 2007. Simultaneous determination of tetracycline, oxytetracycline, and 4-epitetracycline in milk by high-performance liquid chromatography. Food Chem 105:1297–1301. Grabow WOK, Prozesky OW. 1972. Drug Resistance of Coliform Bacteria in Hospital and City Sewage. Antimicrob Agents Chemo 175–180. Gujarathi NP, Haney BJ, Linden JC. 2005. Phytoremediation potential of Myriophyllum Aquaticum and Pista stratiotes to modify antibiotic growth promoters, tetracycline, oxytetracycline, in aqueous wastewater systems. Internatl J Phytorem 7:99–112. Haller MY, Muller SR, McArdell CS, Alder AC, Suter M. 2001. Quantification of veterinary antibiotics (sulfonamids and trimethoprim) in animal manure by liquid chromatography–mass spectrometry. J Chromatogr A 952:111–120. Halling-Sørensen B, Jensen J, Tjørnelund J, Montfors MHMM. 2001. Worst-case estimations of predicted environmental soil concentrations (PEC) of selected veterinary antibiotics and residues in Danish Agriculture. In: Pharmaceuticals in the Environment - Sources, Fate, Effects and Risks, K¨ummerer K (Ed.). New York (NY): Springer. p., 143–156. Halling-Sørensen B, Lykkeberg A, Ingerslev F, Blackwell P, Tjørnelund J. 2003. Characterisation of the abiotic degradation pathways of oxytetracyclines in soil interstitial water using LC-MS-MS. Chemosphere 18;50:1331–1342. Hoagland DR, Arnon DI. 1950. The water-culture method for growing plants without soil, The College of Agriculture, University of California, Berkeley Circular. 347.
Downloaded by [Van Pelt and Opie Library], [Dr Rupali Datta] at 10:25 01 November 2012
PHYTOREMEDIATION OF TETRACYCLINE BY VETIVER GRASS
351
Kasai K, Kanno T, Endo Y, Wakasa K, Tozawa Y. 2004. Guanosine tetra- and pentaphosphate synthase activity in chloroplasts of a higher plant: association with 70S ribosomes and inhibition by tetracycline. Nucleic Acids Res 32:5732–5741. Kemper N. 2008. Veterinary antibiotics in the aquatic and terrestrial environment, Ecological Indicators 8:1–13. Kong WD, Zhu YG, Liang YC, Zhang J, Smith FA, Yang M. 2007. Uptake of oxytetracycline and its phytotoxicity to alfalfa (Medicago sativa L.). Environ Pollut 147:187–193. Kumar K, Gupta SC, Baidoo SK, Chander Y, Rosen CJ. 2005. Antibiotic uptake by plants from soil fertilized with animal manure. J Environ Qual 34:2082–2085. K¨ummerer K. 2009. Antibiotics in the aquatic environment - A review - Part I. Chemosphere 75:417–434. Langhammer JP. 1989. Studies on the fate of antimicrobial drug residues in manure and in the agricultural environment. PhD-Dissertation, University of Bonn. Makris KC, Shakya M, Datta R, Sarkar D, Pachanoor D. 2007. High uptake of 2,4,6-trinitrotoluene by vetiver grass potential for phytoremediation? Environ Pollut 146:1–4. Marcacci S, Raveton M, Ravanel P, Schwitzgu ebel J-P. 2006. Conjugation of atrazine in vetiver (Chrysopogon zizanioides Nash) grown in hydroponics. Env Expt Bot 56:205–215. Meyer MT, Ferrell G, Bumgarner JE, Cole D, Hutchins S, Krapac I, Johnson K, Kolpin D. 2003. Occurrence of antibiotics in swine confined animal feeding operations lagoon samples from multiple states 1998–2002: Indicators of antibiotic use. In: 3rd International Conference on Pharmaceuticals and Endocrine Disrupting Chemicals in Water, National Ground Water Association, Minneapolis. p. 19–21. Migliore L, Cozzolino S, Fiori M. 2003. Phytotoxicity to and uptake of enrofloxacin in crop plants. Chemosphere 52:1233–1244. Peak N, Knapp CW, Yang RK, Hanfelt MM, Smith MS, Aga DS, Graham DW. 2007. Abundance of six tetracycline resistance genes in wastewater lagoons at cattle feedlots with different antibiotic use strategies. Environ Microbiol 9:143–151. Sall J, Creighton L, Lehman A. 2005. JMP Start Statistics, 3rd ed. SAS Institute, Cary, NC/Pacific Grove, CA. Sarmah AK, Meyer MT, Boxall ABA. 2006 A global perspective on the use, sales, exposure pathways, occurrence, fate and effects of veterinary antibiotics (VAs) in the environment, Chemosphere 65:725–759. Scheckler WE, Bennett JV. 1970. Antibiotic usage in seven community hospitals. J Amer Med Asso 213:264–267. Sturtevant AB, Cassell GH, Feary TW. 1971. Incidence of infectious drug resistance among fecal coliforms isolated from raw sewage. Appl Microbiol 21:487–491. Truong P, Van TT, Pinners E. 2008. Vetiver System Applications; Technical Reference Manual. The Vetiver Network International. 2nd Ed. Thomas KV, Dye C, Schlabach M, Langford KH. 2007. Source to sink tracking of selected human pharmaceuticals from two Oslo city hospitals and a wastewater treatment works. J Environ Monitor 9:1410–1418. Watts CD, Craythorne B, Fielding M, Killops SD. 1982. Nonvolatile organic compounds in treated waters. Environ Health Persp 46:87–89.