ResearchersLinks
Advances in Animal and Veterinary SciencesA ResearchersLinks journal
Submit manuscriptReview for this journal

Short Communication · Volume 3, Issue 10, Pages 522–526, 7 August 2015 · Open Access

Extracellular Enzymes and Toxins of <i>Pseudomonas</i> <i>aeruginosa</i> Strains Isolated from Clinically Diseased Egyptian Cows

· · ·

Article Info ⌄
Journal:
Advances in Animal and Veterinary Sciences
Published:
7 August 2015
Download PDF Set Alert Get Rights Reprints

From the article

  • Extracellular Enzymes and Toxins of Pseudomonas Aeruginosa Strains Isolated from Clinically Diseased Egyptian Cows.
  • The obtained results demonstrated that production of high levels of haemolysin (80%), lecithinase (77.1%), protease (34.3%) and elastase enzyme (45.7%) of tested P. aeruginosa strains recovered especially from pneumonia and mastitis infections rather than wound infection.
  • Correlation of proteolytic activity of Pseudomonas aeruginosa with site of isolation.
  • Production and characterization of pseudomonas aeruginosa hemolysin.

Selected from the article's own text.

Abstract

Gamal Younis1, Amal Awad1*, Mona Maghawry2, Fatma Selim1

1Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, 35516; 2Animal Health Research Institute, Dokki, Egypt.

*Correspondence | Amal Awad, Mansoura University, Mansoura, Egypt; Email: dr_amalvet@yahoo.com

Abstract
Pseudomonas aeruginosa produces large array of extracellular toxins that play a crucial role in virulence potential. In this study, the virulence profile of 35 clinical isolates of P. aeruginosa distinctly recovered from pneumonic cattle, mastitic milk samples, and wound infections was determined using phenotypic and genotypic methods. The overall incidence rates from pneumonic cattle, mastitic milk samples, and wound infections were 6%, 25% and 13.3%, respectively with a total incidence of 14%. Recovered isolates were then assayed for their enzymatic activity and the results revealed that out of these 35 isolates, 28 isolates produced haemolytic activity on blood agar plates, 27 isolates showed lecithinase enzyme activity on egg yolk agar media and 12 isolates showed proteolytic enzyme activity on brain heart infusion agar medium supplied with gelatin. Three virulent genes (las B, tox A and exoS) were screened by polymerase chain reaction (PCR) and the results showed that las B, tox A and exoS genes were amplified from 14.7%, 8.57%, and 17.14% of tested isolates, respectively. In conclusion, the present studies confirmed previous observations that the virulence of P. aeruginosa is multifactorial; however, the results indicate that certain exoproducts may play a more important role in certain types of infection.

Keywords | P. aeruginosa, Phenotypic, Genotypic, Virulent genes, PCR

Editor | Kuldeep Dhama, Indian Veterinary Research Institute, Uttar Pradesh, India.

Received | June 11, 2015; Revised | July 29, 2015; Accepted | July 30, 2015; Published | August 07, 2015

Citation | Younis G, Awad A, Maghawry M, Selim F (2015). Extracellular enzymes and toxins of pseudomonas aeruginosa strains isolated from clinically diseased Egyptian cows. Adv. Anim. Vet. Sci. 3(10): 522-526.

ISSN (Online) | 2307-8316; ISSN (Print) | 2309-3331

Copyright © 2015 Younis et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

(M) marker 100pb; Lane 1, 2, 3, 4, 5, 6, 7, 8, 9, 11 and 13: negative; Lane 10, 12: positive; Lane 14: controle negative

(M) marker 100 pb. Lane 1, 2, 3, 4, 5, 6, 7, 8, 9, 11 and12: positive; Lane 10: negative; Lane 7: controle negative

(M) marker 100 pb. Lane 1, 2, 3, 5, 6, 10, 11 and 12: negative; Lane 4, 7, 8, 9: positive; Lane 14: controle negative


Authors’ contributions

All the auhtors contributed equally in research activities and writing this article.

CONFLICT OF INTEREST

The authors state that there ia no conflict of interest.

References

  1. Albert S, Sherry LK, George AO, Zemer G (2014). Surface attachment induces P.aeruginosa virulence. PNAS 111( 47): 16860-16865. http://dx.doi.org/10.1073/pnas.1415712111[CrossRef]
  2. Al-Dujaili AH (1976). Toxic activity against alveolar macrophages of products of Pseudomonas aeruginosa isolated from respiratory and non-respiratory sites. J. Hyg. 77( 2): 211-220. http://dx.doi.org/10.1017/S0022172400024645[CrossRef]
  3. Alina MH, Mariana CC, Ani IC, Coralia B, Alexandru MG, Otilia B, Veronica L (2013). Virulence markers in Pseudomonas aeruginosa isolates from hospitalacquired infections occurred in patients with underlying cardiovascular disease. Rom. Biotech. Lett. 18( 6): 8843-8854.
  4. Amini B, Kamali M, Bayat E, Javadih R, Mansori M, Barhadi N (2010). Isolation and rapid detection pseudomonas aeruginosa by PCR. Quarterly J. Biol. Sci. 3(8): 59-65.
  5. Bever RA, Iglewski BH (1988). Molecular characterization and nucleotide sequence of the Pseudomonas aeruginosa elastase structural gene. J. Bacteriol. 170(9): 4309-4314.
  6. Bodey GP, Bolivar R, Fainstein V, Jadeja L (1983). Infections caused by Pseudomonas aeruginosa. Rev. Infect. Dis. 5(2): 279-313. http://dx.doi.org/10.1093/clinids/5.2.279[CrossRef]
  7. Cruickshank M, McCarteney (1996). Practical microbiology. 123: 413-423.
  8. Deepak B, Lisa S, Hansi K, Kalai M (2013). A dynamic and intricate regulatory network determines P. aeruginosa virulence. Nucleic Acids Res. 41(1): 1–20. http://dx.doi.org/10.1093/nar/gks1039[CrossRef]
  9. Gadeberg OV, Orskov I, Rhodes JM (1983). Cytotoxic effect of an alpha-hemolytic Escherichia coli strain on human blood monocytes and granulocytes in vitro. Infect. Immun. 41(1): 337-341.
  10. Galloway DR, Peters JE (1990). Purification and characterization of an active fragment of the Las A protein from Pseudomonas aeruginosa: enhancement of elastase activity. J. Bacteriol. Mol. Microbiol. 172: 2236-2240.
  11. Habermann E, Hardt KL (1972). A sensitive and specific plate test for the quantitation of phospholipases. Anal. Biochem. 50: 163-173. http://dx.doi.org/10.1016/0003-2697(72)90495-2[CrossRef]
  12. Hanaa ME (1997). Pseudomonas aeruginosa isolated from its possible existence as an etiologic agent of human and animal hazards. M.V.Sc (Vet. Microbiology) Thesis, Faculty of Veterinary Medicine, Zagazig University.
  13. Katarzyne W, Piotr S (2009). Genetic features of clinical Pseudomonas aeruginosa strains. Polish J. Microbiol. 85(3): 255-260.
  14. Khalifa BH, Moissenet D, Vu TH, Khedher M (2011). Virulence factors in Pseudomonas aeruginosa: mechanisms modes of regulation. Ann. Biol. Clin. 69(4): 393-403.
  15. Laila E, Rune B, Stanley J Cryz JR, Bengt W (1986). A comparison of different methods for determining elastase activity of Pseudomonas aeruginosa strains from mink. Acta Path. Microbiol. Immunol. Scand. Sect. B. 94(1-6): 135-138.
  16. Liu PV (1974). Extracellular toxins of Pseudomonas aeruginosa. J. Infect. Dis. 130: S94-S99. http://dx.doi.org/10.1093/infdis/130.Supplement.S94[CrossRef]
  17. Matthew P (1983). The role of exotoxin A in Pseudomonas disease and immunity. Rev. Infect. Dis. 5(5): 979-984.
  18. Matthew P (1984). The Virulence of P. aeruginosa. Clin. Infect. Dis. 6(3): 617-626.
  19. Michael JJ, Sirriatang-nomo EJ, Bottone, Edward P (1980). Correlation of proteolytic activity of Pseudomonas aeruginosa with site of isolation. J. Clin. Microbiol. 12 (4): 626-628.
  20. Nikbin VS, Aslani MM, Sharafi ZH, Ashemipour MS, Ahcheraghi F, Ebrahimipour GH (2012). Molecular identification and detection of virulence genes among Pseudomonas aeruginosa isolated from different infectious origins. Iran. J. Microbiol. 4(3): 118–123.
  21. Pinghui VL (1957). Survey of hemolysin production among species of Pseudomonads. J. Bacteriol. 72: 718-729.
  22. Pinghui VL, Shin Y, Hsieh H (1973). Exotoxins of Pseudomonas aeruginosa. II. Concentration, Purification, and Characterization of Exotoxin A. J. Infect. Dis. 128(4): 514-519. http://dx.doi.org/10.1093/infdis/128.4.514[CrossRef]
  23. Reynolds HY, Fick RB (1979). In “Pseudomonas aeruginosa the organism, diseases its causes and their treatment” (L. D. Sabath, ed.), p. 79. Hans Huber Publishers, Bern.
  24. Richard SB (1962). Production and characterization of pseudomonas aeruginosa hemolysin. J. Bacteriol. 84(5): 1041–1048.
  25. Rocham CL, Coburn J, Rucks EA, Olson JC (2003). Characterization of Pseudomonas aeruginosa exoenzyme S as a bifunctional enzyme in J774A.1 Macrophages. Infect. Immun. 71(9): 5296-5305. http://dx.doi.org/10.1128/IAI.71.9.5296-5305.2003[CrossRef]
  26. Sachin CD, Ruchi K, Santosh S (2012). In vitro evaluation of proteinase, phospholipase and haemolysin activities of Candida species isolated from clinical speciments. Int. J. Med. Biomed. Res. 1(2): 135-157. http://dx.doi.org/10.14194/ijmbr.1211[CrossRef]
  27. Song KP, Chan TK, Ji ZL, Wong SW (2000). Rapid identification of Pseudomonas aeruginosa from ocular isolates by PCR using exotoxin A-specific primers. Mol. Cell. Probes. 14(4): 199-204. http://dx.doi.org/10.1006/mcpr.2000.0306[CrossRef]
  28. Stover C K, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, Brinkman FSL, Hufnagle WO, Kowalik DJ, Lagrou M, Garber RL, Goltry L, Tolentino E, Westbrock-Wadman S, Yuan Y, Brody LL, Coulter SN, Folger KR, Kas A, Larbig K, Lim R, Smith K, Spencer D, Wong GK, Wu Z, Paulsen IT, Reizer J, Saier MH, Hancock REW, Lory S, Olson MV (2000). Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature. 406: 959–964. http://dx.doi.org/10.1038/35023079[CrossRef]
  29. Susumu K, Pinghui VL (1967). Effect of the Hemolysin of Pseudomonas aeruginosa on Phosphatides and on Phospholipase C activity. J. Bacteriol. 93(2): 670-674.
  30. Vermelho AB, Meirelles MN, Lopes A, Petinate SD, Chaia AA, Branquinha MH (1996). Detection of extracellular proteases from microorganisms on agar plates. MemInst Oswaldo Cruz. 91(6): 755-760. http://dx.doi.org/10.1590/S0074-02761996000600020[CrossRef]
  31. Wolfgang MC, Kulasekera BR, Liang X, Boyd D, Wu K, Yang Q, Miyada CG, Lory S (2003). Conservation of genome content and virulence determinants among cilinical and environmental isolates of P.aruginosa. Proceeding of the National Academy of Science of the United States of America. 100: 8484-9.
  32. Wretlind B, Pavlovskis OR (1981). The role of proteases and exotoxin A in the pathogenicity of Pseudomonas aeruginosa infections. Scand. J. Infect. Dis. Suppl. 29: 13-19.

Related articles

Services

Article usage statistics

Loading…

Extracellular Enzymes and Toxins of <i>Pseudomonas</i> <i>aeruginosa</i> Strains Isolated from Clinically Diseased Egyptian Cows · ResearchersLinks