F. Lahig, H., H. Hassan, M., A. Yassir, L. (2018). Molecular Detection of Pseudomonas aeruginosa and Study the Effect of Fresh Garlic juice on Some Virulence Factors of this Bacteria.. , 11(2), 9-17. doi: 10.37652/juaps.2017.141584
Hassan F. Lahig; Muthanna H. Hassan; Luma A. Yassir. "Molecular Detection of Pseudomonas aeruginosa and Study the Effect of Fresh Garlic juice on Some Virulence Factors of this Bacteria.". , 11, 2, 2018, 9-17. doi: 10.37652/juaps.2017.141584
F. Lahig, H., H. Hassan, M., A. Yassir, L. (2018). 'Molecular Detection of Pseudomonas aeruginosa and Study the Effect of Fresh Garlic juice on Some Virulence Factors of this Bacteria.', , 11(2), pp. 9-17. doi: 10.37652/juaps.2017.141584
F. Lahig, H., H. Hassan, M., A. Yassir, L. Molecular Detection of Pseudomonas aeruginosa and Study the Effect of Fresh Garlic juice on Some Virulence Factors of this Bacteria.. , 2018; 11(2): 9-17. doi: 10.37652/juaps.2017.141584

Molecular Detection of Pseudomonas aeruginosa and Study the Effect of Fresh Garlic juice on Some Virulence Factors of this Bacteria.

Journal of University of Anbar for Pure Science
Article 4, Volume 11, Issue 2, 2017, Pages 9-17    PDF (478.97 K)
Document Type: Research Paper
DOI: 10.37652/juaps.2017.141584
Authors
Hassan F. Lahig* 1; Muthanna H. Hassan1; Luma A. Yassir2
1University of Anbar - Collage of Sciences
2Al -Mustansiryah University - National Center of Hematology
Abstract
The present study included the collection of 169 samples from various clinical and environmental sources for investigation of existence of Pseudomonas aeruginosa in those sources, the clinical samples collected from some hospitals in Baghdad city and included 126 samples of wound, burns, cystic fibrosis, ear infections and urinary tract infections (35, 52,4, 22 and 13) samples respectively, while environmental samples were distributed by 25 samples from soil, and 18 samples from wastewater and some swamps. Samples were identified based on the morphological and microscopical characteristics of the colonies when they were culturing in a number of culture media as well as biochemical tests. The molecular detection was used to confirm diagnostic test for the isolates based on the 16SrRNA gene, as a detection gene, which has specific sequences for DNA of P.aeruginosa bacteria.The results of molecular detection showed that 51 isolates belong to target bacteria were distributed as 20 of burns, 9 isolates of wounds, 7 isolates of ear infection, 4 isolates of cystic fibrosis, 2 isolates of urinary tract infection, 5 isolates of soil and 4 isolates of water. The results of phenotypic detection of some virulence factors showed that 49 isolates with percent (95%) produced beta-hemolysis, while 46 isolates (90%) showed variance in the strength of pycocyanin production with a absorbance ranging from (0 to 0.52) at 690 nm, and 47 (92%) of isolates were producing protease enzyme, 44 (86%) of the isolates showed a differences in their ability to biofilm formation. As a result of the increased resistance of P.aeruginosa bacteria towards antibiotics, it is became a problem in the treatment of the infectious diseases caused by this bacteria therefore, the current study focused on the use of fresh garlic extract towards of some virulence factors because it being easy and commonly used. The results of the statistical analysis showed a significant difference when P-value (<0.05) in the production of some virulence factors, the isolates lost their ability to form biofilm, and decreased their ability to produce protease enzyme, and pyocyanin pigment, while did not affect the production of hemolycin.
Keywords
Pseudomonas aeruginosa; Molecular detection; Garlic; virulence factors
References
  1. Kenneth Todar (2011). Pseudomonas, Todar’s online textbook of Bacteriology.
  2. Senturk , S.S.U and Gulgun, B.T. A. and Ulusoy ,S .(2012). Quorum sensing and virulence of Pseudomonas aeruginosa during urinary tract infections. J Infect Dev Ctries . 6(6):501
  3. Kayser, F.H.; Bienz, K.A.; Johannes, E. and Zinkernagel,  R.M. (2005). Kayser, Medical Microbiology, 10thEdition.
  4. Kumar,S. (2012) . vishwe ,M. Katoch. text book of microbiology. 1stED.gaypee brothers medical bublisher. Gellatly, S.L. and Hancock, R.E. (2013).  Pseudomonas aeruginosa: New insights into pathogenesis and host defenses.Pathog. Dis, 67(3):159-73.
  5. Gellatly, S.L. and Hancock, R.E. (2013).  Pseudomonas aeruginosa: New insights into pathogenesis and host defenses. Pathog. Dis, 67(3):159-73.
  6. Govan,J.R.(2007).Pseudomonas aeruginosa and non fermenters ,Medical Microbiology . 17thed . Churchill Livingtone Elsevier.
  7. Lidija Izrael-  Zivkovic; Gordana Gojgic-Cvijovic and Ivanka Karadzi(2010). Isolation and partial characterization of protease from Pseudomonas aeruginosa ATCC 27853. J. Serb. Chem. Soc. 75:1041-1052. 
  8. Abdulnasser, S.S. Ibrahim; Nefisa M. A. El-Shayeb and Sohair, S. Mabrouk (2007). Isolation and identification of alkaline protease producing alkaliphilic bacteria from an Egyption Sadolake. Journal of applied Scinces research. 11:1363-1368.
  9. Ulhusa Patil and Ambala Chaudhari(2011).  Optimal production of alkaline protease from solvent-  tolerant alkalophilic  Pseudomonas aeruginosa. MTCC 7926.India Journal of Biotechnology .10: 329-339.
  10. Tiwari, R. P.; Kamaldeep Deol; Parveen Rishi and J. S.Grewal (2002). Factors affecting haemolysin production and congo red binding in Salmonella entrica serovar Typhimurium DT98. J. Med. Mirobiol. 51:503-509.
  11. Donlan, R.M . (2002). Biofilms: Microbial life on surfaces, Emerg Infect Dis.8:881-890.
  12. Trachoo, N.(2003) .Biofilm  and  food  industry. J.Technol.25:807-815
  13. Overman ,P.R.(2000). Biofilm:A new view of plaque, J. Cont. Dent Prac.1: 2-7.
  14. Costerton,W.; Veeh, R.; Shirtliff , M .; Pasmore ,M and Ehrlich, G. (2003) .The  application  of  biofilm science to the study and control of chronic bacterial  infections, J.Clin.Invest.112:1466-1477.
  15. Davey, M.E and'Toole , G.A.(2000). Microbial biofilms : from ecology to molecular genetics. Microbiol. Mol. Biol. Rev . 64: 847-867.
  16. Parsons JF, Greenhagen BT, Shi K, Calabrese K, Robinson H, Ladner JE.( 2007). Structural and functional analysis of the pyocyanin biosynthetic protein PhzM from Pseudomonas aeruginosa. Biochemistry. 2007; 46; 7; p1821-1828.
  17. Cox, CD.(1986). Role of pyocyanin in the acquisition of iron from transferrin. Infection and immunity. 52; 1; p263-270.
  18. Campbell, M .; Mahenthiralingam  ,E .;  and  Speert , D.P.(2000). Evaluation of random amplified polymorphic DNA typing of Pseudomonas aeruginosa. J. Clin. Microbiol .38:4614-5.
  19. Wolfgang, M.C.; Kulasekara, B.R .; Liang , X.;Boyd, D.; Wu, K and Yang Q,et al.(2003).Conservation of genome content and virulencedeterminants among clinical and environmental isolates of Pseudomonas aeruginosa. Proc. Nat. Acad .Sci. USA.100:8484.
  20. Taylor PW.( 2013). Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents;42(3):195–201.
  21. Abubakar, E.-M. (2009). Efficacy of crude extracts of garlic(Allium sativum) against nosocomial Escherichia coli Staphylococcus aureus Streptococcus pneumoniea and Pseudomonas aeruginosa.pdf. Journal of Medicinal Plants Research, 3(4), 179–185.
  22. Mohsenipour, Z. (2015). The effects of Allium sativum extracts on biofilm formation and activities of six pathogenic bacteria. Jundishapur Journal of Microbiology.
  23. Jaber, M.A. and Al-Mossawi, A. (2007). Susceptibility of some multiple resistant bacteria to garlic extract. Afr. J. Biotechnol. 6 (6): 771-776.
  24. Kock, H. P. and Lawsen, L.D. (1996). Garlic: the science and therapeutic application of Allium sativum and related species.2nd edition. Williams and Willins, New York. p.812.
  25. Chand B (2013). Antibacterial effect of garlic (allium sativum) and ginger (zingiber officinale) against staphylococcus aureus, salmonella typhi, escherichia coli and bacillus cereus. J of Microbiol, Biotech and Food Sci.;2(4):2481–91.10.
  26. Al-salihy, S. R., Karwi, Q. G., & Motib, A. S. (2013). Antibacterial activity of Garlic extract on bacteria isolated from teeth, 187(3), 39–44.
  27. Ankri  S,  Mirelman  D. (1999).  Antimicrobial  properties  of  allicin  from garlic. Microbes and Infection 1: 125–129.
  28. Feldberg, R.S.; Chang, S.C. and Kotik, A.N. (1988). In vitro  mechanism of inhibition of bacterial growth by allicin. Antimicrob. Agents Chemother.32: 1763-1768.
  29. Collee, J.G; Miles, R.S. and Watt, B. (1996). Test for the identification of bacteria. In: Makie and Maccartney practical medical microbiology by collee, J.G; Fraser, A.G; Marmion, B.P. and simmous, A. 4th edition, Churchill livingstone. P: 131149.
  30. Ohman, D.E.; Sadoff, J.C. and Iglewski, B.H. (1980). Toxin-Deficient mutant of Pseudomonas aeruginosa 188 Isolation and characterization.infect  Immune 28(3). 399-408.
  31. Parsons JF, Greenhagen BT, Shi K, Calabrese K, Robinson H, Ladner JE.(2007). Structural and functional analysis of the pyocyanin biosynthetic protein PhzM from Pseudomonas aeruginosa. Biochemistry. 46; 7; p1821-1828.
  32. Rewatkar,A.R. and Wadher,B.J.(2013). Staphylococcus aureus and Pseudomonas aeruginosa-Biofilm formation Methods. IOSR.J. of  Pharm. and Biology. Science.; 8(5):36-40.
  33. Ruppé, E.; Hem, S.; Lath, S.; Gautier, V.; Ariey, F.; Sarthou, J.L.; Monchy, D. and Arlet, G.(2009).CTX-M β-Lactamases in Escherichia coli from Community-acquired Urinary Tract Infections.J.  Emerg. Infect. Dis. 15(5): 741-8.
  34. Reuter;  H.  P.,  Kock  and  D.  L., Lawson(1996).  Therapeutic  effects  and applications  in:  garlic:  The  Science and  Therapeutic   Applications  of A llium  sa tiv um  L.  and  related  species. 2nd Ed. Pp.  135 – 212.
  35. Brooks, G.F. ; Carroll, K.C. ; Butel, J.S. ; Morse, S.A. and Mietzner, T.A. (2010) . Medical Microbiology . 25th ed. Mcgraw-HILL companies . New York .
  36. Abdulridha, S. and Alkaabi, G. (2012).  Bacterial Isolates and Their Antibiograms of Burn Wound Infections in Burns Specialist Hospital in Baghdad.  J. Baghdad for Sci. 10(2): 331.
  37. Khalil, M. A. E. ; Sonbol, F. I. ; Mohamed, A. B. and Ali, S. S.(2015). Comparative study of virulence factors among ESβL-producing and nonproducing Pseudomonas aeruginosa clinical isolates. Turk J Med Sci. 45:60 -69.
  38. Onal, S.; Aridogan, B. C.; Gonen, I.; Tas, T. and Kaya S.(2015). Virulence factors of  Pseudomonas aeruginosa  strains isolated from clinical sample and role of Quorum sensing signal molecules in the pathogenesis of the disease. Acta Medica Mediterranea. 31: 851.
  39. Heydari,  S.  and  Eftekhar, F.  (2014). Biofilm Formation and β-Lactamase  Production  in  Burn  Isolates  of  Pseudomonas aeruginosa .Jundishapur J Microbiol . 8(3): e15514.
  40. Saxena, S.; Banerjee, G.; Garg, R. and Singh, M. (2014). Comparative  Study of Biofilm Formation in Pseudomonas aeruginosa Isolates from Patients of Lower Respiratory Tract infection. J.Clin. and Dia.Res.8(5): 9-11.
  41. Tessema B, Mulu A, Kassu A, Yismaw G.(2006). An in vitro assessment of the antibacterial effect of garlic (Allium sativum) on bacterial isolates from wound infections.
  42. Perez-Giraldo C, Cruz-Villalon G, Sanchez-Silos R, Marti-nez-Rubio R, Blanco MT, Gomez-Garcia AC.(2003). In vitro activity of allicin against Staphylococcus epidermidis and influence of sub inhibitory concentrations on biofilm formation. J Appl Microbiol ;95:709–711.
  43. Chhillar,A. K.and Gahlaut, A. (2013). Evaluation of antibacterial potential of plant extracts using resazurin based microtiter dilution assay. Int J Pharm Pharm Sci. 5: 372-376.
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