Special Issues
Isolation and Phylogenetic Analysis of Polyphosphate Accumulating Organisms in Water and Sludge of Intensive Catfish Ponds in the Mekong Delta, Vietnam
American Journal of Life Sciences
Volume 1, Issue 2, April 2013, Pages: 61-71
Received: Mar. 30, 2013; Published: May 30, 2013
Views 4555      Downloads 213
Le Quang Khoi, Tien Giang Applied Research & Technological Science Service Center, Vietnam; Dept. Microbiology Biotechnology, Can Tho City, Vietnam; Science and Technology Department, Tien Giang province, Vietnam; Biotechnology R&D Institute, Can Tho University, Can Tho City, Vietnam
Cao Ngoc Diep, Tien Giang Applied Research & Technological Science Service Center, Vietnam; Dept. Microbiology Biotechnology, Can Tho City, Vietnam; Science and Technology Department, Tien Giang province, Vietnam; Biotechnology R&D Institute, Can Tho University, Can Tho City, Vietnam
Article Tools
Follow on us
Polyphosphate accumulating organisms were isolated from water and sludge samples of intensive catfish ponds in the Mekong Delta, Vietnam. The Results of estimation of intracellular polyphosphate concentration conducted on each of monocultures indicated that the content of intracellular polyphosphate varied from 2 mg/l to 148.1 mg/l after 6 days of incubation in the medium. Of 191 isolates, twenty-one have uptake and store intracellular phosphate from 19.6 to 148.1 mg/l. They have shaped like a rods and short rods or cocci, a few of them were slightly curved or straight or curved rods. The majority of them are gram-positive (76.2%) and the remains are gram-negative. The partial 16S rRNA genes of these isolates were sequenced and compared with bacterial 16S rRNA genes in Genbank using BlastN Program. Phylogenetic tree was constructed on the basic 16S rRNA gene sequences demonstrating the population of high phosphate accumulating bacteria obtained from samples of catfish ponds were affiliated with four major bacterial lineages. Twenty-one bacteria isolates from samples of catfish ponds included in four classes: Bacilli, Actinobacteria, Beta-proteobacteria, Gamma-proteobacteria. The majority of the strains showed excess phosphate accumulation. Strains related to Bacillus sp. were dominant bacteria group constituted up to 52.4% of all identified isolates, but high phosphate accumulating bacteria are Burkholderia vietnamiensis TVT003L within class Beta-proteobacteria, Acinetobacter radioresistens TGT013L within Gamma-proteobacteria and Arthrobacter protophomiae VLT002L within class Actinobacteria. Methyl blue Loeffler’s staning and electron microscopy examination confirmed that the bacteria had stored polyphosphate granules intracellularly.
Polyphosphate Accumulating Organisms, Polyphosphate, Phosphate Removal, Catfish Pond, Bacilli, Actinobacteria
To cite this article
Le Quang Khoi, Cao Ngoc Diep, Isolation and Phylogenetic Analysis of Polyphosphate Accumulating Organisms in Water and Sludge of Intensive Catfish Ponds in the Mekong Delta, Vietnam, American Journal of Life Sciences. Vol. 1, No. 2, 2013, pp. 61-71. doi: 10.11648/j.ajls.20130102.17
N. P. van Zalinge, P. Degen, P. Pongsiri, S. Nuov, J. G. Jensen, V. H. Nguyen, X .Choulamany, "The Mekong River System" In: R. L. Welcomme, T. Petr (Eds.), Proceedings of the Second International Symposium on the Management of Large Rivers for Fisheries,", Volume 1. FAO Regional Office for Asia and the Pacific, Bangkok, Thailand, RAP Publication 2004/16, pp. 335–357.
Sub-Institute of Water Resources Planning, "Analysis of sub-area 10 V basin development plan," Viet Nam National Mekong Committee, Ho Chi Minh City, November 2003, 87 pp.
Lam T. Phan, Tam M. Bui, Thuy T. T. Nguyen, Geoff J. Gooley, Brett A. Ingram, Hao V. Nguyen, Phuong T. Nguyen, S. S. De Silva, "Current status of farming practices of striped catfish, Pangasianodon hypophthalmus in the Mekong Delta, Vietnam," Aquaculture 296, 2009, pp. 227-236.
S. S. De Silva, Brett. A. Ingram, Phuong T. Nguyen, Tam M. Bui, Geoff J. Gooley, Giovanni M. Turchini, "Estimation of Nitrogen and Phosphorus in Effluent from the Striped Catfish Farming Sector in the Mekong Delta, Vietnam," Ambio, Vol. 39(7), published online 2010 July, pp. 504-514.
P. L. Bond, R. Erhart, M. Wagner, J. Keller, L. L. Blackall, "Identification of some of the the major groups of bacteria in efficient anf non-efficient biolical phosphorus removal activated sludge systems," Appl. Environ. Microbiol., Vol. 65(9), 1999, pp. 4077-4088.
T. Mino, M. C. M. Van Loosdrecht and J. J. Heijnen, "Microbiology and biochemistry of the enhanced biological phosphate removal process," Water Res., Vol. 32, 1998, pp. 3193–3207.
A. Oehmen, P. C. Lemos, G. Carvalho, Z. Yuan, J. Keller, L. L. Backall, M. A. M. Reis, "Advances in enhanced biological phosphorus removal: from micro to macro scale," Water Research, Vol. 41, 2007, pp. 2272-2300.
R-J. Seviour, T. Mino, M. Onuki, "The microbiology of biological phosphorus removal in activated sludge systems," FEMS Microbiology Reviews, Vol. 27, 2003, pp. 99-127.
K. Nakamura, K. Masuda., and E. Mikami, "Isolation of a new type of polyphosphate accumulating bacteria and its phosphate removal characristics," Journal of fermentation and bioengineering, Vol. 71, 1991, pp. 258-263.
Y. Ubukata and S. Takii, "Induction ability of excess phosphate accumulation for phosphate removing bacteria," Water Res., Vol. 28, 1994, pp. 247-249.
G. W. Fuhs and M. Chen, "Microbiological basis of phosphate removal in the activated sludge process for treatment of wastewater," Microbial. Ecol., Vol. 2, 1975, pp. 119-138.
G. R. Crocetti, P. Hugenholtz, P. L. Bond, A. Schuler, J. Keller, D. Jenkings, and L. L. Blackall, "Identification of Polyphosphate-Accumulating Organisms and Design of 16S rRNA-Directed Probes for Their Detection and Quantitation," Applied and environmental microbiology, Vol. 66(3), 2000, pp. 1175-1182.
Y. Kong, J. L. Nielsen and P. H. Nielsen, "Identify ecophysiology of uncultured Actinobacterial polyphosphate accumulating organisms in full scale enhanced biological phosphorus removal plants," Applied and Environmental Microbiology, Vol. 71(7), 2005, pp. 4076-4085.
H. T. T. Nguyen, L. V. Quy, A. A. Hansen, J. L. Nielsen and H. Nielsen, "High diversity and abundence of putative polyphosphate accumulating Tetrasphaera related in activated sludge systems," Microbiol Ecol., Vol. 76, 2011, pp. 256-267.
G. Auling, F. Pilz, H-J. Busse, S. Karrash, M. Streichan and G. Schon, "Analysis of the polyphosphate accumulating microflora in phosphorus eliminating, anaerobic-aerobic activated sludge systems by using Diaminopropane as a biomarker for rapid estimation of Acinetobacter spp," Applied and Environmental Microbiology, 1991, pp. 3585-3592.
M. Sidat, F. Bux and H. C. Kasan, "Polyphosphate accumulation by bacteria isolated from activated sludge," Water SA., Vol. 25(2), 1999, pp. 175-179.
K. D. McMahon, M. A. Dojka, N. R. Pace, D. Jenkins and J. D. Keasling, "Polyphosphate kinase genes from activated sludge carrying outenhanced biological phosphorus removal," Appl. Environ. Microbiol., Vol. 68, 2002, pp. 4971-4978.
S-J. Lee, Y-S. Lee, Y-C. Lee and Y-L. Choi, "Moclecular chracterization of polyphosphate (PolyP) operon from Serratia marcescens," Basic Microbial., Vol. 46(2), 2006, pp. 108-115.
J. M. Kim, H. J. Lee, S. Y. Kim, J. J. Song, W. Park and C. O. Jeon, "Analysis of the Fine-scale population structure of "Candidatus accumulibacter phosphatis" in enhanced biological phosphorus removal sludge, using flurescence in situ hybridization and flow cytometric sorting," Applied and environmental microbiology, Vol. 76(12), 2010, pp.3825-3835.
S. He, D. L. Gall, and K. D. McMahon, "Candidatus Accumulibacter Population Structure in Enhanced Biological Photphorus Removal Sludges as Revealed by Polyphosphate Kinase Genes," Applied and Environmental Microbiology, Vol. 73, 2007, pp. 5865 – 5874.
L. S. Buzoleva, A. M. Krivosheeva, A. S. Isachenko, L. M. Somova and G. P. Somov, "Effect of temperature on synthesis of polyphosphate in Yersinia pseudotuberculosis and Listeria monocytogenes under starvation conditions," Biochemistry (Moscow), Vol. 71(4), 2006, pp. 473-44.
S. Eixler, U. Selig and U. Karsten, "Extraction and detection methods for polyphosphate strorage in autotrophic planktonic organisms," hydrobiologia, Vol. 533(1). 2005, pp. 135-143.
G. J. F. Smolders, J. van der Meij, M. C. M. van Loosdrecht and J. J. Heijnen, "Stoichiometric model ofthe aerobic metabolism of the biological Prusremoval process," Biotech. Bioeng., Vol. 44, 1994, pp. 837-848.
J. Murphy & J. P. Riley, "A modified single solution method for the determination of phosphate in natural waters," Analytica Chimica Acta 27, 1962, pp. 31–36.
C. D. Boswell, R. E. Dick, H. Eccles and L. E. Macaskie, " Phosphate uptake and release by Acinetobacter johnsonii in continuous culture and coupling of phosphate release to heavy metal accumulation," Journal of industrial microbiology and biotechnology, Vol. 26, 2001, pp. 333-340.
E. H. E. Carr, S. Feng, A. Hoogenraad, R. Croome, J. Soddell, K. Lindrea and R. Seviour, "RAPD-PCR typing of Acinetobacter isolates from activated sludge systems designed to remove phosphorus microbiologically," Journal of Applied microbiology, Vol. 90, 2001, pp. 309-319.
D. J. Lane, "16S/23S rRNA sequencing. Nucleic acid techniques in bacterial systematics," E. Stackebrandt and M. Goodfellow, eds. New York, NY, John Wiley and Sons: 115-175, 1991.
V. Ivanov, V. Stabnikov, W. Q. Zhuang, J. H. Tay and S. T. L. Tay, "Phosphate removal from the returned liquor of municipal wastewater treatment plant using iron-reducing bacteria," Applied Microbiology, Vol. 98, 2005, pp. 1152–1161.
K. Tamura, D. Peterson, N. Peterson, G. Stecher, M. Nei and S. Kumar, "MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods," Mol. Biol., Vol. 28(10), 2011, pp. 2731–2739.
G. M. Garrity, "Bergey’s Manual of Systematic Bacteriology, 2nd edition (New York: Springer), 2005.
H. Tamaki, Y. Sekiguchi, S. Hanada, K. Nakamura, N. Nomura, M. Matsumura and Y. Kamagata, "Comparative analysis of bacterial diversity in freshwater dediment of a shallow eutrophic lake by molecular and improved cultivation based techniques," Applied and environmental microbiology, Vol. 71(4), 2005, pp. 2162-2169.
J. Ahn, S. Schroeder, M. Beer, S. Mcllroy, R. C. Bayly, J. W. May, G. Vasiliadis and R. J. Seviour, "Ecology of the Microbial Community removing Phosphate from Wastewater under Continuously Aerobic Conditions in a Sequencing Batch Reactor," Applied and Environmental Microbiology, Vol. 73, 2007, pp. 2257-2270.
S. Gloess, H-P. Grossart, M. Allgaier, S. Ratering and M. Hupfer, "Use of Laser microdissection for phylogenetic characterization of polyphosphate accumulating bacteria," Applied and Environmental Microbiology, Vol. 74(13), 2008, pp. 4231-4235.
H. Ohtake, K. Takahashi, Y. Tsuzuki and K. Toda, "Uptake and release of phosphate by a pure culture of acinetobacter calcoaceticus," Water Res., Vol. 19(12), 1985, pp. 1587-1594.
K. Bark, A. Sponner, P. Kämpfer, S. Grund and W. Dott, "Differences in polyphosphate accumulation and phosphate adsorption by Acinetobacter isolate from wastwater producing polyphosphate: AMP phosphotransferase," Wat. Res., Vol. 26(10), 1992, pp. 1379-1388.
A. Hiraishi, K. Masamune and H. Kitamura, "Characterization of the Bacterial Population Structure in an Anaerobic-Aerobic Activated Sludge System on the Basis of Respiratory Quinone Profiles," Applied and Environmental microbiology, Vol. 55(4), 1989, pp. 897-901.
M. Wagner, R. Erhart, W. Manz, R. Amann, H. Lemmer, D. Wedi and K. H. Schleifer, "Development of an rRNA-targeted oligonucleotide probe specific for the genus Acinetobacter and its application for in situ monitoring in activated sludge," Appl. Environ. Microbiol, Vol. 60(3), 1994, pp. 792-800.
P. L. Bond, P. Hugenholtz, J. Keller And L. L. Blackall, "Bacterial community structure of phosphate removing and non-phosphate removing activated sludges from sequencing batch reactor," Applied and Environmental Microbiology, Vol. 61(5), 1995, pp. 1910-1916.
W-T. Liu, A. T. Nielsen, J-H. Wu, Y. C-S. Tsai, "Matsuo and S. Molin. In situ identification of polyphosphate and polyhydroxyalkanoate accumulating traits for microbial populations in a biological phosphorus removal process," Environmental Microbiology, Vol. 3(2), 2001, pp. 110-122.
G. Szabó, B. Khayer, A. Rusznyák, I. Tátrai, G. Dévai, K. Márialigeti and A K. Borsodi, "Seasonal and spatial variability of sediment bacterial communities inhabiting the large shallow Lake Balaton," Hydrobiologia, Vol. 663, 2011, pp. 217-232.
M. Shoda, T. Ohsumi and S. Udaka, "Screening for high phosphate accumulating bacteria," Agric. Biol. Chem.,Vol. 44(2), 1980, pp. 319-324.
M. Beer, H. M. Stratton, P. C. Griffiths and R. J. Seviour, "Which are the polyphosphate accumulating organisms in full scale activated sludge enhanced biological phosphate removal system in Austalia?," Applied of Microbiology, 100, 2006, pp. 233-243.
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
Tel: (001)347-983-5186