search
for
 About Bioline  All Journals  Testimonials  Membership  News  Donations


Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 24, No. 1, 2016, pp. 38-42
Bioline Code: ej16058
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 24, No. 1, 2016, pp. 38-42

 en A fast and simple assay to quantify bacterial leukotoxin activity
Oppermann, Tobias; Schwarz, Stefan; Busse, Nadine & Czermak, Peter

Abstract

Background: Mannheimia haemolytica check for this species in other resources is the primary bacterial pathogen in causing bovine respiratory disease with tremendous annual losses in the cattle industry. The leukotoxin from M. haemolytica is the predominant virulence factor. Several leukotoxin activity assays are available but not standardized regarding sample preparation and cell line. Furthermore, these assays suffer from a high standard error, a prolonged time consumption and often complex sample pretreatments, which is important from the bioprocess engineering point of view.
Results: Within this study, an activity assay based on the continuous cell line BL3.1 combined with a commercial available adenosine triphosphate viability assay kit was established. The leukotoxin activity was found to be strongly dependent on the sample preparation. Furthermore, the interfering effect of lipopolysaccharides in the sample could be successfully suppressed by adding polymyxin B. We reached a maximum relative P95 value of 14%, which is more than seven times lower compared to current available assays as well as a time reduction up to 88%.
Conclusion: Ultimately, the established leukotoxin activity assay is simple, fast and has a high reproducibility. Critical parameters regarding the sample preparation were characterized and optimized making complex sample purification superfluous.

Keywords
ATP assay; Bacterial pathogen; Bovine respiratory disease; Cattle industry; Critical parameters; Lipopolysaccharides; Mannheimia haemolytica; Microbial biotechnology; Polymyxin B; Quantification; Virulence

 
© Copyright 2016 - Pontificia Universidad Católica de Valparaíso
Alternative site location: http://www.ejbiotechnology.info

Home Faq Resources Email Bioline
© Bioline International, 1989 - 2017, Site last up-dated on 16-Oct-2017.
Site created and maintained by the Reference Center on Environmental Information, CRIA, Brazil
System hosted by the Internet Data Center of Rede Nacional de Ensino e Pesquisa, RNP, Brazil