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Applied and Environmental Microbiology, April 2009, p. 2057-2061, Vol. 75, No. 7
0099-2240/09/$08.00+0     doi:10.1128/AEM.02406-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Effects of Biocides and Other Metal Removal Fluid Constituents on Mycobacterium immunogenum{triangledown}

Joseph O. Falkinham III*

Department of Biological Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0406

Received 20 October 2008/ Accepted 23 January 2009

Cells of Mycobacterium immunogenum are significantly more resistant to biocides and antimicrobial compounds used for disinfection of metal removal fluids (MRFs) than cells of the gram-negative bacterium Pseudomonas pseudoalcaligenes. To ensure accurate measurement of M. immunogenum susceptibility, a biocide inactivation step must be included to prevent an overestimation of killing for short-term exposures. Cell suspensions in 5% MRF for 30 min rather than direct plating following biocide exposure increased the killing of M. immunogenum cells, suggesting a heretofore undiscovered interaction between MRF components and biocides. Biocide killing was reduced at high M. immunogenum cell densities (>106/ml). Among the chemicals tested, only Synergex Premier, Preventol CMK, and dicyclohexylamine killed greater than 90% of M. immunogenum cells in 6 h. M. immunogenum cells adhered to and grew rapidly on glass, copper, and galvanized surfaces, forming biofilms of 106 cells/cm2 within 2 weeks and suggesting it is likely that the majority of M. immunogenum cells in MRF systems are on surfaces. M. immunogenum cells grown on surfaces in biofilms were three- to 100-fold more resistant to the biocides Synergex Premier and Preventol CMK than cells grown in suspension, compounding the problem of eradication of M. immunogenum cells from MRF systems.

* Mailing address: Department of Biological Sciences, Virginia Tech, Blacksburg, VA 24061-0406. Phone: (540) 231-5931. Fax: (540) 231-9307. E-mail: jofiii{at}vt.edu

{triangledown} Published ahead of print on 5 February 2009.

Applied and Environmental Microbiology, April 2009, p. 2057-2061, Vol. 75, No. 7
0099-2240/09/$08.00+0     doi:10.1128/AEM.02406-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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