Abstract
In this study, three adherent Staphylococcus strains able to use nitrate as electron acceptor were isolated from textile wastewater activated sludge. Based on the biochemical profiles, bacterial strains were identified as Staphylococcus lentus, Staphylococcus warneri and Staphylococcus epidermidis the PCR amplification of the nir genes reveal that S. lentus and S. epidermidis were nirK positive and that S. epidermidis was nirS positive. The three strains were also icaA/icaD positive. To obtain the optimal formulation of pure cultures of the staphylococci, the influence of the different mixtures of organisms was studied using mixture design. The regression model of microorganism composition and main metabolites was established. The most predictable reduction of nitrate was 87.2 and 12% of COD consumption. The results suggested that the predictable production of nitrite would reach a minimum of 6.1 mg/l. Based on this, the response values that satisfied all expectations were optimized using MINITAB® 14 analysis software. The most optimal proportion combination was 49.75% of S. lentus (curve value), 36.85% of S. warneri (curve value) and 13.39% S. epidermidis (curve value).
![](https://cdn.statically.io/img/media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-009-0193-1/MediaObjects/11274_2009_193_Fig1_HTML.jpg)
![](https://cdn.statically.io/img/media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-009-0193-1/MediaObjects/11274_2009_193_Fig2_HTML.gif)
![](https://cdn.statically.io/img/media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-009-0193-1/MediaObjects/11274_2009_193_Fig3_HTML.gif)
![](https://cdn.statically.io/img/media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-009-0193-1/MediaObjects/11274_2009_193_Fig4_HTML.gif)
Similar content being viewed by others
References
Arciola CR, Baldassarri L, Montanaro L (2001) Presence of icaa and icad genes and slime production in a collection of staphylococcal strains from catheter-associated infections. J Clin Microbiol 6:2151–2156
Braker G, Fesefeldt A, Witzel KP (1998) Development of PCR primer systems for amplification of nitrite reductase genes (nirK and nirS) to detect denitrifying bacteria in environmental samples. Appl Environ Microbiol 64:3769–3775
Casaburi A, Blaiotta G, Mauriello G, Pepe I, Villani F (2005) Technological activities of Staphylococcus carnosus and Staphylococcus simulans strains isolated from fermented sausages. Meat Sci 71:643–650
Chaieb K, Mahdouani K, Bakhrouf A (2005) Detection of icaA and icaD loci by polymerase chain reaction and biofilm formation by Staphylococcus epidermidis isolated from dialysate and needles in a dialysis unit. J Hosp Infect 61:225–230
Chaieb K, Cheheb O, Zmantar T, Roubhia M, Mahdouani K, Bakhrou A (2007) In vitro effect of pH and ethanol on biofilm formation by clinical ica-positive Staphylococcus epidermidis strains. Ann Microbiol 57(3):431–437
Christensen GD, Simpson WA, Younger JJ, Baddour LM, Barrett FF, Melton DM, Beachey EH (1985) Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 22:996–1006
Coyne MS, Arunakumari A, Averill BA, Tiedje JM (1989) Immunological identification and distribution of dissimilatory heme cd1 and nonheme copper nitrite reductases in denitrifying bacteria. Appl Environ Microbiol 55:2924–2931
Freeman DJ, Falkiner FR, Keane CT (1989) New method for detecting slime production by coagulase-negative staphylococci. J Clin Pathol 42:872–874
Knowles R (1982) Denitrification. Microbiol Rev 46:43–70
Körbahti F, Rauf MA (2008) Application of response surface analysis to the photolytic degradation of Basic Red 2 dye. Chem Eng J 138:166–171
Liu CG, Zhou JB, Chen ZJ (2001) Application of mixture experiment design in studying the effects of fertilizer combination. J Northwest Sci Tech Univ Agric For 29:59–62
Mauriello G, Casaburi A, Blaiotta G, Villani F (2004) Isolation and technological properties of coagulase negative staphylococci from fermented sausages of Southern Italy. Meat Sci 67:149–158
Muteki K, MacGregor JF, Ueda T (2007) Ueda mixture designs and models for the simultaneous selection of ingredients and their ratios. Chemom Intell Lab Syst 8:17–25
Neubauer H, Götz F (1996) Physiology and interaction of nitrate and nitrite reduction in Staphylococcus carnosus. J Bacteriol 178:2005–2009
Nogales B, Timmis KN, Nedwell DB, Osborn AM (2002) Detection and diversity of expressed denitrification genes in estuarine sediments after reverse transcription-PCR amplification from mRNA. Appl Environ Microbiol 68(10):5017–5025
Peters G, Locci R, Pulverer G (1982) Adherence and growth of coagulase-negative staphylococci on surfaces of intravenous catheters. J Infect Dis 146:479–482
Vasudevan P, Nair MK, Annamalai T, Venkitanarayanan KS (2003) Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vet Microbiol 92:179–185
Ward BB (1995) Diversity of culturable denitrifying bacteria. Limits of rDNA RFLP analysis and probes for the functional gene, nitrite reductase. Arch Microbiol 163:167–175
Zhang C, Tong HR, Zhang DM, Li HN (2006) Study on optimization of the formula for vegetable protein drink. J Southwest Agt Univ 28:197–200
Zhou J, Palumbo A, Tiedje JM (1997) Sensitive detection of a novel class of toluene-degrading denitrifiers, Azoarcus tolulyticus, with small-subunit rRNA primers and probes. Appl Environ Microbiol 63:2384–2390
Zmantar T, Chaieb K, Miladi H, Mahdouani K, Bakhrouf A (2006) Detection of the intercellular adhesion loci (ica) in clinical Staphylococcus aureus strains responsible for hospital acquired auricular infection. Ann Microbiol 56(4):349–352
Zorba Ö, Kurt S (2006) Optimization of emulsion characteristics of beef, chicken and turkey meat mixtures in model system using mixture design. Meat Sci 73:611–618
Zumft WG (1992) The denitrifying prokaryotes. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds) The procaryotes, vol 1. Springer, New York, pp 554–582
Zumft WG (1997) Cell biology and molecular basis of denitrification. Microbiol Mol Biol Rev 61:533–616
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Harbi, B., Chaieb, K., Jabeur, C. et al. PCR detection of nitrite reductase genes (nirK and nirS) and use of active consortia of constructed ternary adherent staphylococcal cultures via mixture design for a denitrification process. World J Microbiol Biotechnol 26, 473–480 (2010). https://doi.org/10.1007/s11274-009-0193-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-009-0193-1