PCR Detection of Enterotoxins and Methecillin Resistances Genes in Staphylococcus aureus Isolated from local food in Sulaimania City

Authors

  • Zainab M. Al Zubaidy Salahaddin University/Collage of Agriculture
  • Tazhan M. A. Hama Kareem Sulaimania University/Collage of Agriculture

DOI:

https://doi.org/10.36320/ajb/v10.i1.8197

Keywords:

Staphylococcus aureus , biosynthesis, MERSA

Abstract

Present study was carried out to evaluate bacterial contaminated foods in local markets of Sulaimania city. A total 250 samples were examined for isolation and identification of Staph. aureus, that producer to enterotoxins and resistances to methicillin . The samples included dressing of cake, soft chesses (white), ready to eat foods, red meat, and poultry. The results appeared that: the incidence of Staphylococci were identified in 175 (70%) of the total samples from those; 104(42%) were positive for Staph. aureus which had the ability to grow on the mannitol salt agar media, the most contaminate foods were dressing of cake  51% and soft chess 45% , While red meat was 44% and poultry 48% as well as ready to eat foods which contaminated with 35% of the samples. According to biochemical testes;  Staph. aureus isolates had the ability to produce many enzymes like protease lipase, lethicinase , Coagulase , Catalase, also could fermented mannitol anaerobically and Hemolysin production, while all the isolates were oxidase negative. The production of enterotoxin tested by culture methods for Staph. aureus    isolates showed that 57.14% of the isolates were enterotoxin producer. The  producer isolates were confirmed by PCR to detect the existence of (sea, seb and sec) genes, so 55%of the tested isolates possessed these genes. Sensitivity to antibiotics applied to Staph. aureus isolates that enterotoxin producer revealed different percentage of sensetivity to different antibiotics, the isolates appeared sensitivity toward pencillin, methicillin, vancomycin. The Methicillin resistance Staph. aureus which tested by disk diffusion methods was 81.6%, but by PCR was 73.3%.

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References

Aires de Sousa, M. and Herminia de Lencastre, (2004) “genetic portraits of methicillin-resistant Staphylococcus aureus clones”, FEMS Immunology and Medical Microbiology 40(2):101- 111

Akineden, Ö., Annemüller, C., Hassan, A.A, Lammler, C., Wolter, W., Zschöck M. , (2001) “Toxin genes and other characteristics of Staphylococcus aureus isolates from milk of cows with mastitis”, Clin Diagn Lab Immunol,(8): 959-964.

Balaban, N. And Rasooly, A. 2000. Staphylococcal enterotoxins. Int. J. Food Microbiol. 61,1–10.

Beneke B, Klees S, Stührenberg B, Fetsch A, Kraushaar B, and Tenhagen BA. (2011) “ Prevalence of methicillin- resistant Staphylococcus aureus in a fresh meat pork production chain”, J Food Prot (74):126–129..

Benson. (2001) “Microbiological Applications, Laboratory Manual in General Microbiology

Braun, P., Balzer, G. and Fehlhaber, K. (2001)“Activity of bacterial lipases at chilling temperatures”, J. Food Microbiol. 18: 211- 215.

Brooks, G.F., K.C. Carroll, J.S. Butel, and S.A. Morse ,Jawetz, Melnick and Adelbergs (2007) “Medical Microbiology”. 24th.ed. The McGraw- Hill Companies, Inc., New York.P.224-232.

Cagatay, A.A., Ozcan, P.E., Gulec, L., Ince, N., Tugrul, S., Ozsut, H., Cakar, N., Esen, F., Eraksoy, H. And Calangu, S. 2007. Risk factors for mortality of nosocomial bacteraemia in intensive care units. Med. Princ. Pract. 16, 187–192.

Carroll, K.C., Leonard, R.B., Newcomb-Gayman, P.L. And Hillyard, D.R. 1996. Rapid detection of the staphylococcal mecA gene from BACTEC blood culture bottles by the polymerase chain reaction. Am. J. Clin. Pathol. 106, 600–605.

Cencigoga, B.T., Karama, M., Rossitto, P. V., Morgante, R.A. and Cullor, J. S., (2003) “Enterotoxin production by Staphylococcus aureus isolated from mastitic cows”, J. Food, prot.

Center for Food Safety and Public Health(,CFSPH) Iowa State University. “Methicillin-resistant Staphylococcus aureus ”,;(2011) January. Available at: www.cfsph.iastate.edu/Factsheets/pdfs/mrsa.pdf. Accessed 19 January 2012.

Collee, J.G., Fraser, A.G.M., Marison, B. P. and Simmons, A., (1996) “Mackie and McCarteny Practical Medical Microbiology” 14th.ed. Churchill Livingstone Inc., New York.

Cremonesi, P., Luzzana, M., Brasca, M., Morandi, S., Lodi, R., Vimercati, C., Agnellini, D., Caramenti, G., Moroni, P. And Castiglioni, B. 2005. Development of a multiplex PCR assay”, 8thed, The McGraw-Hill Companies, NewYork. 66(9): 1693-1696.

De Jonge, R., Verdier, J.E, and Havealaar, (2010)“Prevalence of methicillin-resistant Staphylococcus aureus amongst professional meat handlers in the Netherlands”, J. Euro Surveill 15(46):19712.

Dubin, G. (2002)“Extracellular proteases of Staphylococcus spp.”, J. Biol Chem (383):1075- 1086.

Ercolini, D. , Blalotta, G. , Fusco, V., Coppola, S., (2004) “PCR based detection of enterotoxigenic Staphylococcus aureus in the early stages of raw milk cheese making.making”J. Appl. Microbiol 96:1090- 1096.

Foster, T.J. 2005. Immune evasion by staphylococci. Nat. Rev. Microbiol. 3, 948–958.

Francois, P., Pittet, D., Bento, M., Pepey, B., Vaudaux, P., Lew, D. And Schrenzel, J. 2003. Rapid detection of methicillin-resistant Staphylococcus aureus directly from sterile or nonsterile clinical samples by a new molecular assay. J. Clin. Microbiol. 41, 254–260.

Hammer, K.A., Carson, C.F. and Riley, T.V. (2005) “Effects of tea tree oil on Staphylococcus aureus virulence factors” Rural industries research and development corporation. 1- 34. (RIRDC Publication No 05/115, RIRDC Project No UWA 80A).

Harley, J.P. and L. M. Prescott.(2002) “Laboratory Exercises in Microbiology” 5th.ed. The McGraw-Hill Companies, Inc., New York

Holt, J.G.; N.R. Krieg; P.H.A. Sneath; J.T. Staley; and S.T. Williams. (1994) “Bergy,s Manual of Determinative Bacteriology” 9th.ed. Williams &Wilkins, USA.532-551.

Jansen, W.T., Beitsma, M.M., Koeman, C.J., Van Wamel, W.J., Verhoef, J. And Fluit, A.C. 2006. Novel mobile variants of staphylococcal cassette chromosome mec in Staphylococcus aureus. Antimicrob. Agents Chemother. 50, 2072–2078.

Jonas, D., Speck, M., Daschner, F.D. And Grundmann, H. 2002. Rapid PCR-based identification of methicillin-resistant Staphylococcus aureus from screening swabs. J. Clin. Microbiol. 40, 1821–1823.

Jorgensen, H.J., Mork, T., Hogasen, H.R. And Rorvik, L.M. 2005. Enterotoxigenic Staphylococcus aureus in bulk milk in Norway. J. Appl. Microbiol. 99, 158–166.

Kalia, A., Rattan, A. And Chopra, P. 1999. A method for extraction of high-quality and high-quantity genomic DNA generally applicable to pathogenic bacteria. Anal. Biochem. 275, 1–5.

Karahan, M. And Cetinkaya, B. 2006. Coagulase gene polymorphisms detected by PCR in Staphylococcus aureus isolated from subclinical bovine mastitis in Turkey. Vet. J. 174, 428–431.

Karlsson, A. and Arvidson, S. (2002)“Variation in Extracellular Protease Production among Clinical Isolates of Staphylococcus aureus Due to Different Levels of Expression of the Protease Repressor sarA”,J.Infect. Immun.70 (8): 4239- 4246.

Katayama, Y., Zhang, H.Z., Hong, D. And Chambers, H.F. 2003. Jumping the barrier to beta-lactam resistance in Staphylococcus aureus. J. Bacteriol. 185, 5465–5472.

Kusumaningrum, H.D., Riboldi, G., Hazeleger, W.C. and Beumer, R.R. (2003) “Survival of foodborne pathogens on stainless steel surfaces and cross contamination to foods” Int. J. Food

Le Loir Y., Florence B. and Michel G.(2003) Staphylococcus aureus and food poisoning. Genet. Mol. Res. 2 (1): 63-76

Levinson, W. and Jawetz, E. (2000)“Medical microbiology and immunology : Examination and Board Review” 6th ed. LangeMedical book.McGrawH ill companies

Lowy, F.D. 1998. Staphylococcus aureus infections. N. Engl. J. Med. 339,

Matos, J. E., Harmon, R. G.; and Langlois, B. E., (1995) “Lecithinase reaction of Staphylococcus aureus strain of diffrenet origin on Baired- medium”, Appl. Microbiol. Lett. 21: 334-335.

Mc Lauchlin, J.; Narayanan, G.L.; Mithani, V. and O'Neill, G. (2000)“The detection of enterotoxins and toxic shock syndrome toxin genes inStaphylococcus aureus by polymerase chain reaction”, J Food Prot. 63(4):47988.

Mizobuchi, S., Minami, J., Jin, F., Matsushita, O. And Okabe, A. Moon, J.S., Lee, A.R., Kang, H.M., Lee, E.S., Joo, Y.S., Park, Y.H., Nitzsche, S., Zweifel, C. And Stephan, R. 2007. Mobile Genetic Elements And Bacterial Toxinoses: The

Molla B, Byrne M, Abley M, and Gebreyes W. (2010) “Occurrence of methicillin resistant Staphylococcus aureus (MRSA) on-farm, at slaughter and retail pork in commercial swine and implications for food safety” Presentation 93. http:// www.cvmbs.colostate.edu/mip/crwad/2010_Food.htm.

Novick, R.P. And Subedi, A. 2007. The Sapis: Mobile Pathogenicity of methicillin-susceptible Staphylococcus aureus strains at a German University Hospital implicates the circulating-strain pool as a potential source of emerging methicillin-resistant S. aureus clones. J. Clin. Microbiol. 44, 2179–2185.

Oliveira, D.C., Tomasz, A. And De Lencastre, H. 2002. Secrets of Plasmid 49, 93–105.

Omoe, K., Ishikawa, M., Shimoda, Y., Hu, D.L., Ueda, S. And Shinagawa, K. 2002. Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates Harboring seg, seh, or sei genes. J. Clin. Microbiol. 40, 857–862.

Padmapriya, B.P., Ramesh, A., Chandrashekar, A. And Varadaraj, M.C. 2003. Staphylococcal accessory gene regulator (sar) as a signature gene to detect enterotoxigenic staphylococci. J. Appl. Microbiol. 95, 974–981.

Panlilio, A.L., Culver, D.H., Gaynes, R.P., Banerjee, S., Henderson, T.S., Tolson, J.S. And Martone, W.J. 1992. Methicillin resistant Staphylococcus aureus in U.S. hospitals, 1975–1991. Infect.Control Hosp. Epidemiol. 13, 582–586.

Pearson, N. and Smith, T.C. (2011) “The Emergence of Staphylococcus aureus ST398”, Vector Borne Zoo Dis. 11:32

Perez-Roth, E., Claverie-Martin, F., Villar, J. And Mendezalvarez, S. 2001. Multiplex PCR for simultaneous identification of Staphylococcus aureus and detection of methicillin and mupirocin resistance. J. Clin. Microbiol. 39, 4037–4041.

Rode, T. M., Langsrud, S., Holck, A., and Møretrø, T. (2007) “Different patterns of biofilm formation in Staphylococcus aureus under food-related stress conditions”, International Food Microbiology, 16(3):372- 383.

San moon, Jin, Ae Ri Lee, Seung Hyeup Jaw, Mal Nam Kim, Hyun Mi Kang, and Hye Cheong Koo Yi Seok Joo, Yong Ho Park, (2007) “Comparison of antibiogram, staphylococcal enterotoxin productivity, and coagulase genotypes among staphylococcus aureus isolated from animal and vegetable sources in korea”, Journal of food protection, (70) 11: 2541–2548.

Scherrer, D. - Corti, S. - Muehlherr, J.E. - Zweifel, C. - Stephan, R. 2004 “phenotypic and genotypic characteristics of staphylococcus aureus isolates from raw bulk tank milk samples

Seo, Y.H, Jang, J.H., and Moon, K. D. (2010)“Occurrence and characterization of enterotoxigenic Staphylococcus aureus isolated from minimally processed vegetables and sprouts in Korea”, Food Sci Biotechno.19:313– 319

Sharma, N.K., Rees, C.E. And Dodd, C.E. 2000. Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains. Appl. Environ. Microbiol. 66, 1347–1353.

Soriano, J.M.; Font, G.; Rico, H.; Molto, J.C. and Manes, J. (2002) “Incidence of enterotoxigenic staphylococci and their toxins in Foods”. Journal of Food Protection. (65) 5 :857-860.

Tang, J.-N., Shi, X.-M., Shi, C.-L. And Chen, H.-C. 2006. Characterization of a duplex polymerase chain reaction assay for the detection of enterotoxigenic strains of Staphylococcus aureus. J. Rapid Meth. Auto. Microbiol. 10, 201–217.

Thomas, D.Y., Jarraud, S., Lemercier, B., Cozon, G., Echasserieau, K., Etienne, J., Gougeon, M.L., Lina, G. And Vandenesch, F. 2006. Staphylococcal enterotoxin-like toxins U2 and V, two new staphylococcal superantigens arising from recombination within the enterotoxin gene cluster. Infect. Immun. 74, 4724–4734.

Ubukata, K., Nonoguchi, R., Song, M.D., Matsuhashi, M. And Konno, M. 1990. Homology of mecA gene in methicillinresistant Staphylococcus haemolyticus and Staphylococcus simulans to that of Staphylococcus aureus. Antimicrob. Agents Chemother. 34, 170– 172.

Weese, J.S., Avery, BP, and Reid-Smith, R. J.(2010) “Detection and quantification of methicillin-resistant Staphylococcus aureus (MRSA) clones in retail meat products”, Lett Appl

Weigel, L.M., Clewell, D.B., Gill, S.R., Clark, N.C., Mcdougal, L.K., Flannagan, S.E., Kolonay, J.F., Shetty, J., Killgore, G.E. And Tenover, F.C. 2003. Genetic analysis of a high-level vancomycin-resistant isolate of Staphylococcus aureus. Science 302, 1569–1571.

Yang, S., Lin, S., Kelen, G.D., Quinn, T.C., Dick, J.D., Gaydos, C.A. And Rothman, R.E. 2002. Quantitative multiprobe PCR assay for simultaneous detection and identification to species level of bacterial pathogens. J. Clin. Microbiol. 40, 3449–3454.

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Published

2018-03-05

How to Cite

M. Al Zubaidy, Z., & M. A. Hama Kareem, T. (2018). PCR Detection of Enterotoxins and Methecillin Resistances Genes in Staphylococcus aureus Isolated from local food in Sulaimania City. Al-Kufa University Journal for Biology, 10(1), 138–149. https://doi.org/10.36320/ajb/v10.i1.8197

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