Biocontrol of MRSA and E. coli using bacteriophages from cow manure
Abstract
Background: Bacteriophages can be used as biocontrol agents to eliminate the undesirable and targeted bacteria in human beings, environment and industries. We aimed to isolate phages from cow manure and investigate their antibacterial and anti-biofilm formation effect against MRSA and E. coli species.
Method: E. coli and S. aureus isolates of clinical origin were sub-cultured on MacConkey’s and Mannitol salt agar media, respectively. To confirm the Methicillin resistance in S. aureus, mecA gene was detected by the PCR using gene specific forward and reverse primers. The bacteriophages were isolated using enrichment procedure from cow manure. Their antibacterial and anti-biofilm formation activity was determined through the inhibition of bacterial growth and decrease in the absorbance of Crystal Violet through biofilm inhibition assay, respectively.
Results: The isolated phages showed clear plaque formation against MRSA and E. coli species. The time-dependent lytic assay showed a sharp decline in OD600 of MRSA and E. coli after one hour incubation with the phages. Biofilm inhibition was indicated by a decrease in the absorbance (OD595) of CV in a microtiter well plate
Conclusion: This is perhaps the first study to isolate phages from cow manure at local region and demonstrate their lytic efficiency against MRSA and E. coli, with the ultimate aim to use them as a promising biocontrol agent against antibiotic resistant pathogens.
Keywords: Phages; MRSA; Plaque formation; Biofilm inhibition; E. coli; Biocontrol
Full Text:
PDFReferences
Twort FW. An investigation on the nature of ultra-microscopic viruses. The Lancet, (1915); 186(4814): 1241-1243.
Barrangou R, Fremaux, Christophe, Deveau, Hélène, Richards, Melissa, Boyaval, Patrick, Moineau, Sylvain, Romero, Dennis A., Horvath, Philippe. CRISPR Provides Acquired Resistance Against Viruses in Prokaryotes. Science, (2007); 315(5819): 1709-1712.
Henry M, Debarbieux L. Tools from viruses: bacteriophage successes and beyond. Virology, (2012); 434(2): 151-161.
Lederberg EM, Lederberg J. Genetic studies of lysogenicity in Eschericia coli. Genetics, (1953); 38(1): 51-64.
Sulakvelidze A, Alavidze Z, Morris JG, Jr. Bacteriophage therapy.
Antimicrob Agents Chemother, (2001); 45(3): 649-659.
Joerger RD. Alternatives to antibiotics: bacteriocins, antimicrobial peptides and bacteriophages. Poultry science, (2003); 82(4): 640-647.
Juhász-Kaszanyitzky E, Jánosi S, Somogyi P, Dán A, van der Graaf-van Bloois L, et al. MRSA transmission between cows and humans. Emerging infectious diseases, (2007); 13(4): 630-632.
Cuny C, Wieler LH, Witte W. Livestock-Associated MRSA: The Impact on Humans. Antibiotics (Basel, Switzerland), (2015); 4(4): 521-543.
Ramírez-Castillo FY, Moreno-Flores AC, Avelar-González FJ, Márquez-Díaz F, Harel J, et al. An evaluation of multidrug-resistant Escherichia coli isolates in urinary tract infections from Aguascalientes, Mexico: cross-sectional study. Annals of clinical microbiology and antimicrobials, (2018); 17(1): 34-34.
Azizian Reza AJF, Razieh Amini5, Karimi Sajedeh,Sadeghifard Norkhoda, Sekawi Zamberi, Nasser Ahmad, Taherikalani Morovat, Askari Hassanand Pakzad Iraj. Dynamics of Bacteriophages as a Promising Antibiofilm Agents. JOURNAL OF PURE AND APPLIED MICROBIOLOGY,, (2014); 8(2): p. 1015-1019.
O'Toole G, Kaplan HB, Kolter R. Biofilm formation as microbial development. Annual review of microbiology, (2000); 5449-79.
Curtin JJ, Donlan, R. M. Using bacteriophages to reduce formation of catheter-associated biofilms by Staphylococcus epidermidis. Antimicrob Agents Chemother, (2006); 50(4): 1268-1275.
Kevin A. Hughes IWSaMVJ. Biofilm susceptibility to bacteriophage attack :the role of phage-borne polysaccharide depol yrnerase. Microbiology, (1998); 1443039-3047.
Sambrook JR, D. W. . Molecular Cloning: A Laboratory Manual,NY, USA (2001). Cold Spring Harbor Laboratory Press: New York,, (2012); 4th ed.
Chhibber S, Kaur T, Kaur S. Essential role of calcium in the infection process of broad-spectrum methicillin-resistant Staphylococcus aureus bacteriophage. Journal of Basic Microbiology, (2014); 54(8): 775-780.
Jurczak-Kurek A, Gasior T, Nejman-Falenczyk B, Bloch S, Dydecka A, et al. Biodiversity of bacteriophages: morphological and biological properties of a large group of phages isolated from urban sewage. Sci Rep, (2016); 634338.
S.A. DOSS GSTaSGBA. Effect of sub-inhibitory concentrations of antibiotics on the virulence of Staphylococcus aureus. Journal of Applied Bacteriology, (1993); 75123-128.
Callaway TR, Edrington TS, Brabban AD, Keen JE, Anderson RC, et al. Fecal Prevalence of Escherichia coli O157, Salmonella,Listeria, and Bacteriophage Infecting E. coli O157:H7 inFeedlot Cattle in the Southern Plains Region of theUnited States. FOODBORNE PATHOGENS AND DISEASE, (2006); 3(3).
Synnott AJ, Kuang Y, Kurimoto M, Yamamichi K, Iwano H, et al. Isolation from sewage influent and characterization of novel Staphylococcus aureus bacteriophages with wide host ranges and potent lytic capabilities. Appl Environ Microbiol, (2009); 75(13): 4483-4490.
Rodrigo-Troyano A, Sibila O. The respiratory threat posed by multidrug resistant Gram-negative bacteria. Respirology, (2017); 22(7): 1288-1299.
Cahill J, Young, R. Phage Lysis: Multiple Genes for Multiple Barriers. Advances in Virus Research, (2019); 10333-70.
Stewart PS. Mechanisms of antibiotic resistance in bacterial biofilms. International journal of medical microbiology : IJMM, (2002); 292(2): 107-113.
Hall-Stoodley L, Costerton JW, Stoodley P. Bacterial biofilms: from the Natural environment to infectious diseases. Nature Reviews Microbiology, (2004); 2(2): 95-108.
Neguţ AC, Chifiriuc M-C, Săndulescu O, Streinu-Cercel A, Oprea M, et al. Bacteriophage-driven inhibition of biofilm formation in Staphylococcus strains from patients attending a Romanian reference center for infectious diseases. FEMS Microbiology Letters, (2016); 363(18).
Kjelleberg S, Molin S. Is there a role for quorum sensing signals in bacterial biofilms? Current Opinion in Microbiology, (2002); 5(3): 254-258.
DOI: http://dx.doi.org/10.62940/als.v7i4.976
Refbacks
- There are currently no refbacks.