Asian Journal of Pharmacy and Technology

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Ambroxol Blocks Swarming and Swimming Motilities and Inhibits Biofilm formation by Proteus mirabilis Isolated from Diabetic Foot Infection


Affiliations
1 Department of Microbiology and Immunology-Faculty of Pharmacy-Zagazig University- Zagazig- Egypt, Egypt
     

   Subscribe/Renew Journal


The aim of this study was to investigate the ability of ambroxol to block the swarming and swimming motilities and biofilm formation of Proteus mirabilis isolated from diabetic foot ulcers as a new agent for the treatment of diabetic foot infections.

Assays of swarming and swimming motilities on LB agar in the presence and absence of sub-inhibitory concentrations of ambroxol were performed by measuring the distance of swarming and swimming in addition to microscopical examination of vegetative and swarmer cells. Moreover, biofilm formation and removal by ambroxol was investigated in polystyrene microtiter plates.

Ambroxol in sub-inhibitory concentrations significantly blocked swarming and swimming motilities in a dosedependent manner. Ambroxol decreased swarming and swimming at concentrations of 0.6, 0.7, 0.8 and 0.9 mg/ml. At a concentration of 0.9 mg/ml, ambroxol completely inhibited swarming and swimming. Ambroxol also could significantly inhibit biofilm formation and remove pre-formed biofilms in a concentration-dependent manner.

This study suggests that ambroxol could be used for the treatment of Proteus mirabilis diabetic foot infections due to its ability to interfere with swarming and invasion of tissues in addition to inhibition of biofilm formation and removal of established biofilms.


Keywords

Proteus mirabilis, Ambroxol, Diabetic Foot Infection, Swarming, Swimming, Biofilm Inhibition
Subscription Login to verify subscription
User
Notifications
Font Size


  • Singh N, Armstrong DG and Lipsky BA. Preventing foot ulcers in patients with diabetes. Journal of American Medical Association. 2005; 293:217–228.

  • Alavi SM et al. Bacteriologic study of diabetic foot ulcer. Pakistan Journal of Medical Sciences. 23; 2007:681–684.

  • Anandi C et al. Bacteriology of diabetic foot lesions. Pakistan Journal of Medical Sciences. 22; 2004:175–178.

  • Gadepalli R et al. A Clinico-microbiological study of diabetic foot ulcers in an Indian tertiary care hospital. Diabetes Care. 29; 2006:1727–1732. 5. Allison C et al. The role of swarm cell differentiation and multicellular migration in the uropathogenicity of Proteus mirabilis. Journal of Infectious Diseases. 169; 1994:1155–1158.

  • Allison C and Hughes C. Closely linked genetic loci required for swarm cell differentiation and multicellular migration by Proteus mirabilis. Molecular Microbiology. 5; 1991:1975–1982.

  • Rauprich O et al. Periodic phenomena in Proteus mirabilis swarm colony development. Journal of Bacteriology. 178; 1996:–6538.

  • Costerton, JW, Stewart PS and Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science. 284; 1999:1318–1322.

  • Pratt LA and Kolter R. Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Molecular Microbiology. 30; 1998:285–293.

  • Mah TF and O’Toole GA. Mechanisms of biofilm resistance to antimicrobial agents. Trends in Microbiology. 9; 2001:34–39.

  • Stewart PS. Mechanisms of antibiotic resistance in bacterial biofilms. International Journal of Medical Microbiology. 292; 2002:107–113.

  • James G et al. Biofilms in Chronic wounds. Wound Repair and Regeneration. 16; 2008:37–44.

  • O’Toole GA and Kolter R. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Molecular Microbiology. 30; 1998:295–304.

  • Sauer K et al. Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. Journal of Bacteriology. 184; 2002:1140–1154.

  • Stoodley P et al. Biofilms as complex differentiated communities. Annual Review of Microbiology. 56; 2002:187–209.

  • Yamada T et al. Action of N-acylated ambroxol derivatives on secretion of chloride ions in human airway epithelia. Biochemical and Biophysical Research Communications. 380; 2009:586– 590.

  • Stetinová V, Herout V and Kvetina J. In vitro and in vivo antioxidant activity of ambroxol. Clinical and Experimental Medicine. 4; 2004:152–158.

  • Hafez MM et al. Activity of some mucolytics against bacterial adherence to mammalian cells. Applied Biochemistry and Biotechnology. 158; 2009:97–112.

  • Lu Q et al. Ambroxol interferes with Pseudomonas aeruginosa quorum sensing. International Journal of Antimicrobial Agents. 36; 2010:211–215.

  • Koneman EW et al. Colour atlas and textbook of diagnostic microbiology. Lippincott, Philadelphia. 1997, 5th ed: pp. 253–320.

  • Clinical and Laboratory Standards Institute Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard. 7th ed. CLSI Document M7-A7. Wayne, PA. 2006.

  • Hay NA et al. A nonswarming mutant of Proteus mirabilis lacks the Lrp global transcriptional regulator. Journal of Bacteriology. 179; 1997:4741–4746.

  • Stepanovic S et al. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods. 40; 2000:175–179.

  • Viswanathan V and Kumpatla S. Pattern and causes of amputation in diabetic patients – A multicentric Study from India. Journal of the Association of Physicians of India. 59; 2011:148–151.

  • Pecoraro RE, Reiber GE and Burgess EM. Pathways to diabetic limb amputation. Basis for prevention. Diabetes Care. 13; 1990:513–521.

  • Harley JR. Preventing diabetic foot diseases. Nurse Practice. 18; 1993:37–38, 41–44.

  • Loing P. Diabetic foot ulcers. American Journal of Surgery. 167; 1994:31S-36S.

  • Rózalski A, Sidorczyk Z and Kotelko K. Potential virulence factors of Proteus bacilli. Microbiology and Molecular Biology Reviews. 61; 1997:65–89.

  • Manos J and Belas R. The genera Proteus, Providencia and Morganella. In The Prokaryotes: A Hand Book on the Biology of Bacteria, Edited by Dworkin M et al. New York, Springer Science+Business Media. 2006; 3rd ed: pp. 245–269.

  • Belas R and Suvanasuthi R. The ability of Proteus mirabilis to sense surfaces and regulate virulence gene expression involves FliL, a flagellar basal body protein. Journal of Bacteriology. 187; 2005:6789–6803.

  • Stickler DJ and Zimakoff J. Complications of urinary tract infections associated with devices used for long-term bladder management. Journal of Hospital Infection. 28; 1994:177–194.

  • Donlan RM and Costerton JW. Biofilms:survival mechanisms of clinically relevant microorganisms. Clinical Microbiology Reviews. 15; 2002:167–193.

  • Li F et al. Effects of ambroxol on alginate of mature Pseudomonas aeruginosa biofilms. Current Microbiology. 57; 2008:1–7.

  • Abbas HA, Serry FM and EL-Masry EM. Combating Pseudomonas aeruginosa biofilms by potential biofilm inhibitors. Asian Journal of Research in Pharmaceutical Sciences. 2; 2012:66–72.

  • Wang WB et al. Inhibition of swarming and virulence factor expression in Proteus mirabilis by resveratrol. Journal of Medical Microbiology. 55; 2006:1313–1321.

  • Liaw SJ et al. Inhibition of virulence factor expression and swarming differentiation in Proteus mirabilis by pnitrophenylglycerol. Journal of Medical Microbiology. 49; 2000:725–731.

  • Chow S et al. Salicylic acid affects swimming, twitching and swarming motility in Pseudomonas aeruginosa, resulting in decreased biofilm formation Journal of Experimental Microbiology and Immunology. 15; 2011:22–29.

  • Daniels R, Vanderleyden J and Michiels J. Quorum sensing and swarming migration in bacteria. FEMS Microbiology Reviews. 28; 2004:261–289.


Abstract Views: 978

PDF Views: 0




  • Ambroxol Blocks Swarming and Swimming Motilities and Inhibits Biofilm formation by Proteus mirabilis Isolated from Diabetic Foot Infection

Abstract Views: 978  |  PDF Views: 0

Authors

Hisham A. Abbas
Department of Microbiology and Immunology-Faculty of Pharmacy-Zagazig University- Zagazig- Egypt, Egypt

Abstract


The aim of this study was to investigate the ability of ambroxol to block the swarming and swimming motilities and biofilm formation of Proteus mirabilis isolated from diabetic foot ulcers as a new agent for the treatment of diabetic foot infections.

Assays of swarming and swimming motilities on LB agar in the presence and absence of sub-inhibitory concentrations of ambroxol were performed by measuring the distance of swarming and swimming in addition to microscopical examination of vegetative and swarmer cells. Moreover, biofilm formation and removal by ambroxol was investigated in polystyrene microtiter plates.

Ambroxol in sub-inhibitory concentrations significantly blocked swarming and swimming motilities in a dosedependent manner. Ambroxol decreased swarming and swimming at concentrations of 0.6, 0.7, 0.8 and 0.9 mg/ml. At a concentration of 0.9 mg/ml, ambroxol completely inhibited swarming and swimming. Ambroxol also could significantly inhibit biofilm formation and remove pre-formed biofilms in a concentration-dependent manner.

This study suggests that ambroxol could be used for the treatment of Proteus mirabilis diabetic foot infections due to its ability to interfere with swarming and invasion of tissues in addition to inhibition of biofilm formation and removal of established biofilms.


Keywords


Proteus mirabilis, Ambroxol, Diabetic Foot Infection, Swarming, Swimming, Biofilm Inhibition

References