International Journal of Environmental Bioremediation & Biodegradation
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International Journal of Environmental Bioremediation & Biodegradation. 2014, 2(5), 220-227
DOI: 10.12691/ijebb-2-5-2
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Isolation and Characterization of Linuron Degrading Bacteria from Soils under Horticultural Production in Kenya

Philip Miriti1, , Gabriel Magoma.2, Hamadi I. Boga3 and Aggrey Nyende B1

1Institute of Biotechnology Research Jomo Kenyatta University of agriculture and Technology P.O Box 62000 (00200) Nairobi, Kenya

2Pan African University Institute of Basic Science Technology And Innovation Jomo Kenyatta University of agriculture and Technology – AICAD Block C, Room 101 P. O Box 62000 00200 Nairobi, Kenya

3Hamandi Iddi, Jomo Kenyatta University of agriculture and Technology P.O Box 62000 (00200) Nairobi, Kenya

Pub. Date: September 08, 2014

Cite this paper:
Philip Miriti, Gabriel Magoma., Hamadi I. Boga and Aggrey Nyende B. Isolation and Characterization of Linuron Degrading Bacteria from Soils under Horticultural Production in Kenya. International Journal of Environmental Bioremediation & Biodegradation. 2014; 2(5):220-227. doi: 10.12691/ijebb-2-5-2


Pesticides use has been one of the major factors in improving productivity in agricultural enterprises in Kenya. Phenylurea herbicides are one of the main categories of crop protection products that kill weeds and other plants that grow where they are unwanted. In recent years researchers have been paying greater attention to this family of herbicides because of their high biotoxicity and possible carcinogenic properties and lengthy periods of time for their removal from the environment. However, systematic studies on microbial removal of these pesticide residues are scarce in developing countries including Kenya. A survey in four horticultural regions of central and rift-valley Kenya showed that out of the twenty two (22) formulations of pesticides used linuron had the highest frequency of application at 45.9%. Through enrichment and isolation a total of six isolates were obtained from linuron contaminated soils. Degradation kinetics of the pesticides residues was monitored by High pressure liquid chromatography (HPLC). Level of degradation was scored by the data fit with a linear regression by the line y =49508x with an R2 value of 0.9831. The corroboration of various degradation intermediate metabolites was aided by GC-MS machine. The identified metabolites from linuron were 3, 4 dichloroanilline, 3,-chloroanalline, 4-chloroanalline N, O dimethylhydroxylamine, and N-methyl-N-methoxy carbamic acid methyl ester. Isolates designated LoG-8A, Lwa-2A, Lsh-6B, LJk-5C, Lla-1A and Lwa-2C showed the ability to degrade 70.7- 98.9% of 50mgl-1 of linuron within 84 days. Morphological, biochemical and 16S rRNA sequence analysis of these isolates indentified them with the genus Pseudomonas, Stenotrophomanas, Burkholderia, Lysinibacillus, Arthrobacter and Flavobacteria. The study demonstrated that soils from horticultural farms in Kenya harbor biodegrades for linuron. Further work can show whether these microbes can be used for the development of bioremediation strategy.

pesticides biotoxicity bioremediation Linuron and contamination

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[1]  Amine-Khodja., A. Boulkamh, A. and Boule., P. “Photochemical behavior of phenylurea herbicides.” Photochemical and photobiology science. 3: 145-156. 2004.
[2]  Anthony, R., Smith, W., and Carol, A, “Induction of enzymes of 2, 4 dichlorophenoxyacetate degradation in Burkholderia cepacia 2a and toxicity of metabolic intermediates”, Biodegradation., 19, (5). 669-681. Sep. 2008.
[3]  Alexander, K. Steve., K., and Strete. D, Microbiology: a photographic atlas for the laboratory. Benjamin Cummings publishers, California, 2001.
[4]  Benitez, F.J., Acero, J.L., Real, F.J., and Garcia, C, “Removal of phenyl-urea herbicides in ultrapure water by ultrafiltration and nanofiltration processes”. Water Research, 43 (2) 267-276. Feb 2009.
[5]  Birech, R., Bernhard, F., and Joseph, M.. “Towards reducing synthetic pesticide imports in favour of locally available botanicals in Kenya” in the Conference on International Agricultural Research for Develop, Bonn Germany 8-12.
[6]  Boon, N., Goris, J., De Vos, P., Verstraete, W., and Top, E. “Genetic diversity among 3-chloroaniline-and aniline-degrading strains of the Comamonadaceae”. Appllied and Environmental Microbiology. 67 (3) 1107-1115. Mar 2001.
[7]  Caux, P.Y., Kent, R. A., Fan, G. T., and Grande, C. “Canadian water quality guidelines for Linuron.” Environ mental Toxicololgy and Water Quality 13 (1). 1-41. Dec 1998.
[8]  Cullington, J., and Walker A. “A rapid biodegradation of diuron and other phenylurea herbicides by a soil bacterium”. Soil Biolology and Biochemistry. 31 (5). 677-686. May 1999.
[9]  Dejonghe, W., Goris J., Dierickx, A., De Dobbeleer, V., Crul, V., De Vos, P., Verstraete, W., and Top, E. “Diversity of 3-chloroaniline and 3,4-dichloroaniline degrading bacteria isolated from three different soils and involvement of their plasmids in chloroaniline degradation.” FEMS microbiology and ecology. 42 (2). 315-325. Jan 2006.
[10]  Devers, M. Azhari, N.E. Udikovic Kolic, N. and Martin Laurent, F. Detection and organization of atrazine degrading genetic potential of seventeen bacterial isolates belonging to divergent taxa indicate a recent common origin of their catabolic functions, FEMS Microbiology Letters 273 (1) 78-86. June 2007.
[11]  Gregersen T, “Rapid method for distinction of Gram-negative from Gram-positive Bacteria”. European Journal of applied Microbial Biotechnology 5 (2) 123-127. 1978
[12]  Gooddy, D.C., Mathias S.A., Harrison I., Lapworth. D.J. and Kim A.W, “The significance of colloids in the transport of pesticides through Chalk.” Science Total Environment. 385 (1-3) 262-271. Oct 2007.
[13]  Guzzella, L., Capri, L., DiCorcia, A., Barra Caracciolo, A. and Guiliano, G. “Fate of Diuron and Linuron in a Field Lysimeter Experiment.” Journal of Environmental Quality 35: 312-323. 2006.
[14]  Hiroaki, I., Kazuya, A. and Yoshie, H. “Isolation and characterization of a new 2, 4-dinitrophenol-degrading bacterium Burkholderia sp. strainKU-46 and its degradation pathway.” FEMS Microbiology Letters, 274 (1). 112-117. Sep. 2007.
[15]  Holt, J. G., Krieg, N. R., Sneath, P. H. A., Staley, J. T. and Williams, S. T. Bergey’s Manual of Determinative Bacteriology, 9th Ed. Williams and Wilkins publishers, London, 1994.
[16]  Johnson, A. Besien, T. Bhardwaj, C. Dixon A, Gooddy, D. Haria, A. and White, C. “Penetration of herbicides to groundwater in an unconfined chalk aquifer following normal soil applications.” Contaminant hydrology. 53: 101-117. 2001.
[17]  Lambright, C., Ostby J., Bobsiene K., Wilson V., Hotchkiss K., Mann C. and Gray E, “ Cellular and Molecular Mechanisms of action of Linuron an Antiandrogenic Herbicide that Produces Malformations in Male Rats”. Toxicological Science, 56 (2). 389-399. Aug 2000.
[18]  Lamoree, M., Swart, C., van der Horst, A. and van Hattum B. “Determination of diuron and the antifouling paint biocide Irgarol1051 in Dutch marinas and coastal waters.” Journal of Chromatography A, 970: 183-190, 2002.
[19]  Macharia, I., Mithofer, D.and Waibel, H. “Potential environmental impacts of pesticides use in the vegetable sub-sector in kenya.” African Journal of Horticultural Science 2, 138-151. 2009.
[20]  Marmur, J. “A procedure for the isolation of deoxyribonucleic acid from micro-organisms.” Journal of Molecular Biolology 3: 208-218. Jan 1961.
[21]  Mehrdad, E., Wabule, M.N., Ngaruiya, P.N., Kimmins, F.K. and Silverside, P.J. “Problems facing the flower industry and registration for Biocontrol Agents in Kenya,” in Proceedings of the PCPB/KARI/DFID CPP Workshop, Nakuru, Kenya, 14-16 May 2003.
[22]  Muhammad, S., Asif, T., Muhammad, A., Khuram, M., Naeem, S., Muhammad, I. and Imran, Q. “Effect of weed-crop competition on growth and yield of garden cress Lepidium sativum L”. Journal of Medicinal Plants Research. 5 (26), 6169-6172. Nov 2011.
[23]  Philip, B., Pieter-Jan, D., Rene De, M., and Dirk, S. “Characterization of novel linuron-mineralizing bacterial consortia enriched from long-term Linuron-treated agricultural soils.” FEMS microbial and ecology 62: 374-385. Nov 2007.
[24]  Polz, M. and Cavanaugh, C. “Bias in template-to-product ratios in multitemplate PCR.” Applied Environmental Microbiology 64 (10) 3724-3730. Oct 1998.
[25]  PCPB (2005). Pest Control Products Board, July, 2004-June, 2005 Annual Report.
[26]  Ramanathan, M. and Lalithakumari, D. “Complete mineralization of methylparathion by Pseudomonas sp. A3”. Applied biochemistry and biotechnology. 80 (1) 1-12. April 1999.
[27]  Ritter, R. Scheringer, M. MacLeod, M. Moeckel, C. Jones, K. and Hungerbühler K. “Intrinsic human elimination half lives of polychlorinated biphenyls derived from the temporal evolution of cross-sectional biomonitoring data from the UK population.” Environmental Health Perspect 119: 225-231.2011
[28]  Richardson, M. “Pesticides—friend or foe?” Water Science and Technology 37 (8) 19-25. August 1998.
[29]  Scassellati-Sforzolini, G. Pasquini, R. Moretti, M. Villarini, M. Fatigoni, C. Dolara, P. Monarca, S. Caderni, G. Kuchenmeister, F. Schmezer, P. and B. L. Pool-Zobel. “In vivo studies on genotoxicity of pure and commercial linuron. Mutat. Res. Genet. Toxicol.” Environ mental Mutagen. 390 (3) 207-22. 1997.
[30]  Smith, D. Alvey, S. and Crowley, D. “Cooperative catabolic pathways within an atrazine degrading enrichment culture isolated from soil.’ FEMS Microbiology and Ecology. 53 (2): 265-273. Jul 2005.
[31]  Sorensen, S.R., Bending, G.D., Jacobsen, C.S., Walker, A. and Aamand, J. “Microbial degradation of isoproturon and related phenylurea herbicides in and below agricultural fields.” FEMS Microbiology of Ecology, 45 (1) 1-11. July 2003.
[32]  Surovtseva, E. G. Sukhikh, A. P. and Ivoilov, V. S. “Isozymes of the pathway for aniline and 4-chloroaniline preparatory metabolism in Alcaligenes sp. Mikrobiologiy, 61: 99-106. 1993
[33]  Tamura, K. Dudley, J. Nei, M. and Kumar, S. “MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0.” Molecular Biology and Evolution 24 (8): 1596-1599. August 2007
[34]  Thompson, J. Higgins, D. and Gibson, T. “CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.” Oxford journal of nucleic acid research. 22 (22) 4673-4680. Sep 1994.
[35]  Tixier., C. Sancelme., M. Bonnemoy., F. Cuer., A. and Veschambre., H. “ Degradation products of a phenylurea herbicide, diuron: synthesis, ecotoxicity, and biotransformation.” Environmental Toxicology & Chemistry, 20: 1381-1389, 2001.
[36]  U.S. Environmental Protection Agency (U.S. EPA) (1995) R.E.D. Facts Linuron (EPA-738-F-95-003). pp. 1-11 (
[37]  Uyttebroek, M. Breugelmans, P. Janssen, M. Wattiau, P. Joffe, B. Karlson, U. Ortega-Calvo, JJ. Bastiaens, L. Ryngaert, A. Hausner, M. Springael, D. “Distribution of the Mycobacterium community and polycylic aromatic hydrocarbons (PAHs) among different size fractions of a long-term PAH-contaminated soil.” Environmental Microbiology 8 (5) 836-847. May 2006.
[38]  Ware, G.W. and Whitacre, D.M. The Pesticide Book, MeisterPro Information Resources, Willoughby, Ohio. pp. 496.
[39]  Widehem P, Ait-Aissa S, Tixier C, Sancelme M, Veschambre H and Truffaut, N. “ Isolation, characterization and diuron transformation capacities of a bacterial strain Arthrobacter sp. N2.” Chemoshere, 46 (4) 527-534. Jan 2002.