Abstract
The floating filter technique was successfully adapted for the isolation of the dominant, chemolithoautotrophic, sulfide-oxidizing bacterium from a sulfur-producing reactor after conventional isolation techniques had failed. The inoculated polycarbonate filters, floating on mineral medium, were incubated under gaseous hydrogen sulfide at non-toxic levels. This technique gave 200-fold higher recoveries than conventional isolation techniques. Viable counts on the filters, making up 15% of the total count, appeared to be all of the same species. Chemostat cultures of the new isolate had a very high sulfur-forming capacity, converting almost all hydrogen sulfide in the medium to elemental sulfur under high sulfide loads (27.5 mmol l-1 h-1) and fully aerobic conditions. This behaviour closely resembled that of the microbial community in the sulfur-producing reactor. Moreover, similar protein patterns were obtained by electrophoresis of cell-free extracts from the isolate and the mixed culture. It has therefore been concluded that this isolate represents the dominant sulfide-oxidizing population in the reactor. The isolate has been shown to be a new Thiobacillus species, related to Thiobacillus neapolitanus. In view of the general confusion currently surrounding the taxonomy of the thiobacilli, a new species has not been formally created. Instead, the isolate has been given the working name Thiobacillus sp. W5.
Similar content being viewed by others
References
Bartlett JK & Skoog DA (1954) Colorimetric determination of elemental sulphur in hydrocarbons. Anal. Chem. 26: 1008–1011
de Bruyn JC, Boogerd FC, Bos P & Kuenen JG (1990) Floating filters, a novel technique for isolation and enumeration of fastidious, acidophilic, iron-oxidizing, autotrophic bacteria. Appl. Environ. Microbiol. 56: 2891–2894
Buisman CJN, Ijspeert P, Geraats S & Lettinga G (1989) Biotechnological process for sulphide removal with sulphur reclamation. Acta. Biotechnol. 9: 271–283
Buisman CJN, Ijspeert P, Hof A, Janssen AJH, ten Hagen R & Lettinga G (1991) Kinetic parameters of a mixed culture oxidizing sulfide and sulfur with oxygen. Biotechn. Bioeng. 38: 813–820
Coates JD, Phillips EJP, Lonergan DJ, Jenter H & Lovley DR (1996) Isolation of Geobacter species from diverse sedimentary environments. Appl. Environ. Microbiol. 62(5): 1531–1536
Fox GE, Wisotzkey JD & Jurtshuk P (1992) How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int. J. Syst. Bacteriol. 42(1): 166–170
Gottschal JC & Kuenen JG (1980) Selective enrichment of facultatively chemolithotrophic thiobacilli and related organisms in the chemostat. FEMS Microbiol. Lett. 7: 241–247
Hazeu W, Batenburg-van der Vegte WH, Bos P, van der Plas RK & Kuenen JG (1988) The production and utilization of intermediary elemental sulphur during the oxidation of reduced sulphur compounds by Thiobacillus ferrooxidans. Arch. Microbiol. 150: 574–579
Hirayama A & Vetter RD (1989) Kinetics of sulphide and thiosulphate oxidation by the hydrothermal vent Bacterium Thiomicrospira crunogina and comparison with Thiobacillus neapolitanus. Abstract I43. Proceedings of the annual meeting-1989 American Society of Microbiology, Washington DC
Katayama-Fujimura Y, Tsuzaki N, Kuraishi H (1982) Ubiquinone, fatty acid and DNA base composition determination as a guide to the taxonomy of the genus Thiobacillus. J. Gen. Microbiol. 128: 1599–1611
Kelly DP, Cambers LA & Trüdinger PA (1969) Cyanolysis and spectrophotometric estimation of trithionate in mixture with thiosulphate and tetrathionate. Anal. Chem. 41: 899–901
Kelly DP & Harrison AP (1989) The genus Thiobacillus. In Bergey's Manual of Systematic Bacteriology, pp 1842–1858. Edited by JP Staley, MP Bryant, N Pfennig & JG Holt. Baltimore, Williams & Wilkins
Kersters K & de Ley J (1980) Classification and identification of bacteria by electrophoresis of their proteins. In Microbiological classification and identification, pp 273–297. Edited by m Good-fellow & RG Board, London, Academic Press
Kuenen JG & Tuovinen OH (1981) The genera Thiobacillus and Thiomicrospira. In The Prokaryotes, pp. 1023–1036.. Edited by MP Starr, H Stolp, HG Trüper, A Balows and HG Schlegel. New York, Springer-Verlag
Kuenen JG, Robertson LA & Tuovinen OH (1992) The genera Thiobacillus, Thiomicrospira and Thiosphaera. In The Prokaryotes, Vol III, pp 2636–2657. Edited by A Balows, HG Trüper, M Dworkin, W Harder, & KH Schlefer. New York, Springer-Verlag
Laemmli UK (1970) Cleavage of proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685
Lane DJ, Harrison AP, Stahl D, Pace B, Giovannoni SJ, Olsen GJ & Pace R (1992) Evolutionary relationships among sulphur-and iron-oxidizing eubacteria. J. Bact. 174(1): 269–278
de Ley J, Cattoir H & Reynaerts A (1970) The quantitative measurement of DNA hybridization rates. Eur. J. Biochem. 12: 133–142
Matin A (1978) Organic nutrition of chemolitotrophic bacteria. Ann. Rev. Microbiol 32: 433–469
Martinez-Murcia AJ, Benlloch S & Collins MD (1992) Phylogenetic interrelationships of members of the genera Aeromonas and Pleisomonas as determined by 16S ribosomal DNA sequencing: Lack of congruence with results of DNA-DNA hybridizations. Int. J. Syst. Bacteriol. 42(3): 412–421
Rainey FA, Dorsch M, Morgan W & Stackebrandt E (1992) 16S rDNA analysis of Spirochaeta thermophila: position and implications for the systematics of the order Spirochaetales. System. Appl. Microbiol. 16: 224–226
Rainey FA & Stackebrandt E (1993) 16S rDNA analysis reveals phylogenetic diversity amongst the polysaccharolytic clostridia. FEMS Microbiol. Lett. 133: 125–128
Rand MC, Greenberg AE & Taras MJ (1985) Standard methods for the examination of water and wastewater, 16th edition., APHA-AWWA-WPCF, Washington
Sokolova GA & Karavaiko GI (1968) Physiology and geochemical activity of thiobacilli. Translated from Russian (1964), E Rabinovitz (Ed.). Jerusalem: Israel programme for Scientific Translations Ltd.
Stefess GC (1993) Oxidation of sulphide to elemental sulphur by aerobic thiobacilli. PhD thesis, Delft University of Technology, The Netherlands
Stefess GC, Torremans RAM, de Schrijver R, Robertson LA & Kuenen JG (1996) Quantitative measurement of sulphur formation by steady-state and transient-state continuous cultures of autotrophic Thiobacillus species. Appl. Microbiol. Biotechnol. 45: 169–175
Tichy R, Janssen A, Grotenhuis JTC, Lettinga g & Rulkens WH (1994) Possibilities for using biologically-produced sulphur for cultivation of thiobacilli with respect to bioleaching processes. Bioresource Technology 48: 221–227
Trüper HG & Schlegel HG (1964) Sulphur metabolism in thiorhodaceae: Quantitative measurements on growing cells of Chromatium okenii. Ant. v. Leeuwenhoek. 30: 225–238
Verduyn C, Postma E, Scheffers WA & van Dijken JP (1990) Physiology of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures. J. Gen. Microbiol. 136: 395–403
Vishniac W & Santer M (1957) The Thiobacilli. Bacteriol. Rev. 21: 195–213
Visser JM, de Jong GAH, Robertson LA & Kuenen JG (1997) Purification and characterization of a periplasmic thiosulfate dehydrogenase from the obligately autotrophic Thiobacillus sp. W5. Arch. Microbiol. 166: 372–378
Wayne LG, Brenner DJ, Colwell RR, Grimont PAD, Kandler O, Krichevsky MI, Moore LH, Moore WEC, Murray RGE, Stackebrandt E, Starr MP & Trüper HG (1987) Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. Int. J. Syst. Bacteriol. 37(4): 463–464
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Visser, J.M., Stefess, G.C., Robertson, L.a. et al. Thiobacillus sp. W5, the dominant autotroph oxidizing sulfide to sulfur in a reactor for aerobic treatment of sulfidic wastes. Antonie Van Leeuwenhoek 72, 127–134 (1997). https://doi.org/10.1023/A:1000252126252
Issue Date:
DOI: https://doi.org/10.1023/A:1000252126252