AiM  Vol.6 No.4 , April 2016
Studies on Enterotoxins and Antimicrobial Resistance in Staphylococcus aureus Isolated from Various Sources
Abstract: Staphylococcus aureus represents a public health challenge all over the world. Therefore, this study aims to analyze the prevalence of five genes (sea, seb, sec, see and seg) encoding the staphylococcal enterotoxins in S. aureus isolated from different sources and to evaluate the association of these toxins in comparison to susceptibility towards 12 antimicrobials; antimicrobial susceptibility was conducted by disc diffusion method. Detection of staphylococcal enterotoxins was performed by PCR and the ability to express these genes was assessed among isolates by RT-PCR. The most common enterotoxin gene was sea gene (66%), followed by seb, sec, see and seg (38%, 23%, 19% and 5%) respectively. Expression of sea, seb and seg genes was variable. However, sec and see genes were not expressed by any of the tested isolates. No statistically significant association exists between (seb, sec and see) and isolation sources, while the sea was significantly associated with clinical isolates. High significant correlation was found between elevated sea expression and multidrug-resistance. Our findings indicate that the pathogenic potential of S. aureus may be greater than previously thought. This emphasizes the utmost need to implement proactive measures and more emphasis will be placed on the application of hygiene practices in hospitals to control S. aureus infection and enterotoxins production.
Cite this paper: Mohammed, E. , Abdel-Rhman, S. , Barwa, R. and El-Sokkary, M. (2016) Studies on Enterotoxins and Antimicrobial Resistance in Staphylococcus aureus Isolated from Various Sources. Advances in Microbiology, 6, 263-275. doi: 10.4236/aim.2016.64026.

[1]   Harris, L.G., Foster, S.J. and Richards, R.G. (2002) An Introduction to Staphylococcus aureus, and Techniques for Identifying and Quantifying S. aureus Adhesins in Relation to Adhesion to Biomaterials: Review. European Cells and Materials Journal, 4, 39-60.

[2]   Plata, K., Rosato, A.E. and Wegrzyn, G. (2009) Staphylococcus aureus as an Infectious Agent: Overview of Biochemistry and Molecular Genetics of Its Pathogenicity. Acta Biochimica Polonica, 56, 597-612.

[3]   Heyman, D.L. (2004) Control of Communocable Diseases Manual. American Public Health Association, Washington DC.

[4]   Ortega, E., Abriouel, H., Lucas, R. and Galvez, A. (2010) Multiple Roles of Staphylococcus aureus Enterotoxins: Pathogenicity, Superantigenic Activity, and Correlation to Antibiotic Resistance. Toxins (Basel), 2, 2117-2131.

[5]   El-Jakee, J., Nagwa, A.S., Bakry, M., Zouelfakar, S.A., Elgabry, E. and El-Said, W.G. (2008) Characteristics of Staphylococcus aureus Strains Isolated from Human and Animal Sources. American-Eurasian Journal of Agricultural & Environmental Sciences, 4, 221-229.

[6]   Jackranukul, C., Panvisavas, N. and Chaturongakul, S. (2013) Detection of Staphylococcus aureus Enterotoxin Type I Using qPCR. Chiang Mai Journal of Science, 40, 143-150.

[7]   Jorgensen, H.J., Mork, T., Hogasen, H.R. and Rorvik, L.M. (2005) Enterotoxigenic Staphylococcus aureus in Bulk Milk in Norway. Journal of Applied Microbiology, 99, 158-166.

[8]   Nema, V., Agrawal, R., Kamboj, D.V., Goel, A.K. and Singh, L. (2007) Isolation and Characterization of Heat Resistant Enterotoxigenic Staphylococcus aureus from a Food Poisoning Outbreak in Indian Subcontinent. International Journal of Food Microbiology, 117, 29-35.

[9]   Principato, M. and Qian, B.F. (2014) Staphylococcal Enterotoxins in the Etiopathogenesis of Mucosal Autoimmunity within the Gastrointestinal Tract. Toxins (Basel), 6, 1471-1489.

[10]   Balaban, N. and Rasooly, A. (2000) Staphylococcal Enterotoxins. International Journal of Food Microbiology, 61, 1-10.

[11]   Omoe, K., Hu, D.-L., Ono, H.K., Shimizu, S., Takahashi-Omoe, H., Nakane, A., Uchiyama, T., Shinagawa, K. and Imanishi, K. (2013) Emetic Potentials of Newly Identified Staphylococcal Enterotoxin-Like Toxins. Infection and Immunity, 81, 3627-3631.

[12]   Ono, H.K., Sato’o, Y., Narita, K., Naito, I., Hirose, S., Hisatsune, J., Asano, K., Hu, D.L., Omoe, K., Sugai, M. and Nakane, A. (2015) Identification and Characterization of a Novel Staphylococcal Emetic Toxin. Applied and Environmental Microbiology, 81, 7034-7040.

[13]   Fernandez, M.M., De Marzi, M.C., Berguer, P., Burzyn, D., Langley, R.J., Piazzon, I., Mariuzza, R.A. and Malchiodi, E.L. (2006) Binding of Natural Variants of Staphylococcal Superantigens SEG and SEI to TCR and MHC Class II Molecule. Molecular Immunology, 43, 927-938.

[14]   Pinchuk, I.V., Beswick, E.J. and Reyes, V.E. (2010) Staphylococcal Enterotoxins. Toxins (Basel), 2, 2177-2197.

[15]   Munson, S.H., Tremaine, M.T., Betley, M.J. and Welch, R.A. (1998) Identification and Characterization of Staphylococcal Enterotoxin Types G and I from Staphylococcus aureus. Infection and Immunity, 66, 3337-3348.

[16]   Blaiotta, G., Ercolini, D., Pennacchia, C., Fusco, V., Casaburi, A., Pepe, O. and Villani, F. (2004) PCR Detection of Staphylococcal Enterotoxin Genes in Staphylococcus spp. Strains Isolated from Meat and Dairy Products. Evidence for New Variants of seG and seI in S. aureus AB-8802. Journal of Applied Microbiology, 97, 719-730.

[17]   Chiang, Y.C., Liao, W.W., Fan, C.M., Pai, W.Y., Chiou, C.S. and Tsen, H.Y. (2008) PCR Detection of Staphylococcal Enterotoxins (SEs) N, O, P, Q, R, U, and Survey of SE Types in Staphylococcus aureus Isolates from Food-Poisoning Cases in Taiwan. International Journal of Food Microbiology, 121, 66-73.

[18]   Collee, J.G., Miles, R.S. and Watt, B. (1996) Tests for the Identification of Bacteria. Mackie and Macartney Practical Medical Microbiology, Churchill Livingstone, New York, 413-424.

[19]   Clinical and Laboratory Standards Institute (2011) Performance Standards for Antimicrobial Susceptibility Testing; Twenty-First Informational Supplement. M100-S21. Clinical and Laboratory Standards Institute, Wayne.

[20]   Alfatemi, S.M., Motamedifar, M., Hadi, N. and Saraie, H.S. (2014) Analysis of Virulence Genes among Methicillin Resistant Staphylococcus aureus (MRSA) Strains. Jundishapur Journal of Microbiology, 7, e10741.

[21]   Brakstad, O.G., Aasbakk, K. and Maeland, J.A. (1992) Detection of Staphylococcus aureus by Polymerase Chain Reaction Amplification of the nuc Gene. Journal of Clinical Microbiology, 30, 1654-1660.

[22]   Abdel-Rhman, S.H. (2016) Evaluation of Total RNA Isolation Methods from Staphylococcus aureus. New Egyptian Journal of Microbiology, 43, 50-57.

[23]   Schmittgen, T.D. and Livak, K.J. (2008) Analyzing Real-Time PCR Data by the Comparative CT Method. Nature Protocols, 3, 1101-1108.

[24]   Verkaik, N.J., Dauwalder, O., Antri, K., Boubekri, I., de Vogel, C.P., Badiou, C., Bes, M., Vandenesch, F., Tazir, M., Hooijkaas, H., Verbrugh, H.A., van Belkum, A., Etienne, J., Lina, G., Ramdani-Bouguessa, N. and van Wamel, W.J. (2010) Immunogenicity of Toxins during Staphylococcus aureus Infection. Clinical Infectious Diseases, 50, 61-68.

[25]   Murray, P.R., Baron, E.J., Jorgenson, J.H., Landry, M.L., Pfaller, M.A. and Yolken, R.H. (2003) Manual of Clinical Microbiology. American Society of Microbiology Press, Washington DC.

[26]   Arvidson, S. and Tegmark, K. (2001) Regulation of Virulence Determinants in Staphylococcus aureus. International Journal of Medical Microbiology, 291, 159-170.

[27]   Sabouni, F., Mahmoudi, S., Bahador, A., Pourakbari, B., Sadeghi, R.H., Ashtiani, M.T., Nikmanesh, B. and Mamishi, S. (2014) Virulence Factors of Staphylococcus aureus Isolates in an Iranian Referral Children’s hospital. Osong Public Health and Research Perspectives, 5, 96-100.

[28]   Zigo, F., Vasi, M., Elecko, J., Farkasová, Z. and Chripková, M. (2014) Antibiotic Resistance and Production of Enterotoxins of Staphylococcus spp. Isolated from Samples of Milk in Dairy Sheep. International Journal of Latest Research in Science and Technology, 3, 99-102.

[29]   Ippolito, G., Leone, S., Lauria, F.N., Nicastri, E. and Wenzel, R.P. (2010) Methicillin-Resistant Staphylococcus aureus: The Superbug. International Journal of Infectious Diseases, 14, S7-S11.

[30]   Abulreesh, H.H. (2011) Multidrug-Resistant Staphylococci in the Environment. International Conference on Biotechnology and Environment Manual, 18, 1-6.

[31]   Manoharan, M., Jayaraman, S., Illanchezian, S. and Jayasekar, J. (2008) Homologous Resistant Groups (HRGs) in Multidrug Resistant Staphylococcus aureus Isolated from Retail Foods in Vadapalani, Chennai. The Internet Journal of Microbiology, 7.

[32]   Sina, H., Ahoyo, T.A., Moussaoui, W., Keller, D., Bankole, H.S., Barogui, Y., Stienstra, Y., Kotchoni, S.O., Prevost, G. and Baba-Moussa, L. (2013) Variability of Antibiotic Susceptibility and Toxin Production of Staphylococcus aureus Strains Isolated from Skin, Soft Tissue, and Bone Related Infections. BMC Microbiology, 13, 188.

[33]   Tokajian, S., Haddad, D., Andraos, R., Hashwa, F. and Araj, G. (2011) Toxins and Antibiotic Resistance in Staphylococcus aureus Isolated from a Major Hospital in Lebanon. International Scholarly Research Network Microbiology, 2011, Article ID: 812049.

[34]   Perveen, S., Naqvi, S.B. and Fatima, A. (2013) Antimicrobial Susceptibility Pattern of Clinical Isolates from Cases of Ear Infection Using Amoxicillin and Cefepime. SpringerPlus, 2, 288.

[35]   Opere, B.O., Fashola, M.O., Adesida, S., Smith, S.I. and Adebiyi, O.A. (2013) Prevalence and Antibiotic Susceptibility Pattern of Staphylococcus aureus in Clinical Specimens. Advances in Life Science and Technology, 10, 1-5.

[36]   Schmitz, F.J., Fluit, A.C., Gondolf, M., Beyrau, R., Lindenlauf, E., Verhoef, J., Heinz, H.P. and Jones, M.E. (1999) The Prevalence of Aminoglycoside Resistance and Corresponding Resistance Genes in Clinical Isolates of Staphylococci from 19 European Hospitals. The Journal of Antimicrobial Chemotherapy, 43, 253-259.

[37]   Elazhari, M., Elhabchi, D., Zerouali, K., Dersi, N., Elmalki, A., Hassar, M., Saile, R. and Timinouni, M. (2011) Prevalence and Distribution of Superantigen Toxin Genes in Clinical Community Isolates of Staphylococcus aureus. Journal of Bacteriology & Parasitology, 2, 107.

[38]   Shittu, A.O. and Lin, J. (2006) Antimicrobial Susceptibility Patterns and Characterization of Clinical Isolates of Staphylococcus aureus in KwaZulu-Natal Province, South Africa. BMC Infectious Diseases, 6, 125.

[39]   Hauschild, T., Sacha, P., Wieczorek, P., Zalewska, M., Kaczynska, K. and Tryniszewska, E. (2008) Aminoglycosides Resistance in Clinical Isolates of Staphylococcus aureus from a University Hospital in Bialystok, Poland. Folia Histochemica et Cytobiologica, 46, 225-228.

[40]   Gade, N.D. and Qazi, M.S. (2014) Recent Trend of Aminoglycoside Resistance among Staphylococcus aureus Isolates in Tertiary Care Hospital. Journal of Microbiology and Antimicrobials, 6, 94-96.

[41]   Rayner, C. and Munckhof, W.J. (2005) Antibiotics Currently Used in the Treatment of Infections Caused by Staphylococcus aureus. Internal Medicine Journal, 35, S3-S16.

[42]   Asadollahi, P., Delpisheh, A., Maleki, M.H., Jalilian, F.A., Alikhani, M.Y., Asadollahi, K., Soroush, S., Hematian, A., Emaneini, M. and Taherikalani, M. (2014) Enterotoxin and Exfoliative Toxin Gtenes among Methicillin-Resistant Staphylococcus aureus Isolates Recovered from Ilam, Iran. Avicenna Journal of Clinical Microbiology and Infection, 1, e20208.

[43]   Fernandes, C.J., Fernandes, L.A. and Collignon, P. (2005) Cefoxitin Resistance as a Surrogate Marker for the Detection of Methicillin-Resistant Staphylococcus aureus. The Journal of Antimicrobial Chemotherapy, 55, 506-510.

[44]   Yaseen, I.H., Shareef, A.Y. and Daoud, A.S. (2013) High Prevalence of Multidrug-Resistance MRSA and VRSA of Different Infections from Al-Jumhuory Teaching Hospital Patients in Mosul. Journal of Life Sciences, 7, 1255-1259.

[45]   Seas, C., Hernandez, K., Ramos, R., Bazan, E., Rodriguez, I., Torres, A., Zamudio, C. and Gotuzzo, E. (2006) Oxacillin-Resistant and Multidrug-Resistant Staphylococcus aureus in Lima, Peru. Infection Control and Hospital Epidemiology, 27, 198-200.

[46]   Pillar, C.M., Draghi, D.C., Sheehan, D.J. and Sahm, D.F. (2008) Prevalence of Multidrug-Resistant, Methicillin-Resistant Staphylococcus aureus in the United States: Findings of the Stratified Analysis of the 2004 to 2005 Leader Surveillance Programs. Diagnostic Microbiology and Infectious Disease, 60, 221-224.

[47]   Cao, H., Wang, M., Zheng, R., Li, X., Wang, F., Jiang, Y. and Yang, Y. (2012) Investigation of Enterotoxin Gene in Clinical Isolates of Staphylococcus aureus. Journal of Southern Medical University, 32, 738-741.

[48]   Becker, K., Friedrich, A.W., Lubritz, G., Weilert, M., Peters, G. and Von Eiff, C. (2003) Prevalence of Genes Encoding Pyrogenic Toxin Superantigens and Exfoliative Toxins among Strains of Staphylococcus aureus Isolated from Blood and Nasal Specimens. Journal of Clinical Microbiology, 41, 1434-1439.

[49]   Naffa, R.G., Bdour, S.M., Migdadi, H.M. and Shehabi, A.A. (2006) Enterotoxicity and Genetic Variation among Clinical Staphylococcus aureus Isolates in Jordan. Journal of Medical Microbiology, 55, 183-187.

[50]   Kelly, B.G., Vespermann, A. and Bolton, D.J. (2009) Horizontal Gene Transfer of Virulence Determinants in Selected Bacterial Foodborne Pathogens. Food and Chemical Toxicology, 47, 969-977.

[51]   Argudin, M.A., Mendoza, M.C. and Rodicio, M.R. (2010) Food Poisoning and Staphylococcus aureus Enterotoxins. Toxins (Basel), 2, 1751-1773.

[52]   Bhatty, M., Ray, P., Singh, R., Jain, S. and Sharma, M. (2013) Presence of Virulence Determinants amongst Staphylococcus aureus Isolates from Nasal Colonization, Superficial & Invasive Infections. The Indian Journal of Medical Research, 138, 143-146.

[53]   Indrawattana, N., Sungkhachat, O., Sookrung, N., Chongsa-nguan, M., Tungtrongchitr, A., Voravuthikunchai, S.P., Kong-ngoen, T., Kurazono, H. and Chaicumpa, W. (2013) Staphylococcus aureus Clinical Isolates: Antibiotic Susceptibility, Molecular Characteristics, and Ability to form Biofilm. BioMed Research International, 2013, Article ID: 314654.

[54]   Kamarehei, F., Ghaemi, E.A. and Dadgar, T. (2013) Prevalence of Enterotoxin A and B Genes in Staphylococcus aureus Isolated from Clinical Samples and Healthy Carriers in Gorgan City, North of Iran. Indian Journal of Pathology & Microbiology, 56, 265-268.

[55]   Safaei, H.R., Pirasteh, H., Pournasiri, Z. and Dormanesh, B. (2015) Study the Enterotoxigenixity of Staphylococcus aureus Isolated from the Urine Samples of Pediatrics with UTIs. Biomedical and Pharmacology Journal, 8, 111-118.

[56]   Wang, L.X., Hu, Z.D., Hu, Y.M., Tian, B., Li, J., Wang, F.X., Yang, H., Xu, H.R. and Li, Y.C. (2013) Molecular Analysis and Frequency of Staphylococcus aureus Virulence Genes Isolated from Bloodstream Infections in a Teaching Hospital in Tianjin, China. Genetics and Molecular Research, 12, 646-654.

[57]   Lim, K.T., Hanifah, Y.A., Mohd Yusof, M.Y. and Thong, K.L. (2012) Investigation of Toxin Genes among Methicillin-Resistant Staphylococcus aureus Strains Isolated from a Tertiary Hospital in Malaysia. Tropical Biomedicine, 29, 212-219.

[58]   Adwan, G., Adwan, K., Jarrar, N., Salama, Y. and Barakat, A. (2013) Prevalence of seg, seh and sei Genes among Clinical and Nasal Staphylococcus aureus Isolates in Palestine. British Microbiology Research Journal, 3, 139-149.

[59]   Varshney, A.K., Mediavilla, J.R., Robiou, N., Guh, A., Wang, X., Gialanella, P., Levi, M.H., Kreiswirth, B.N. and Fries, B.C. (2009) Diverse Enterotoxin Gene Profiles among Clonal Complexes of Staphylococcus aureus Isolates from the Bronx, New York. Applied and Environmental Microbiology, 75, 6839-6849.

[60]   Sapri, H.F., Sani, N.A.M., Neoh, H.-M. and Hussin, S. (2013) Epidemiological Study on Staphylococcus aureus Isolates Reveals Inverse Relationship between Antibiotic Resistance and Virulence Repertoire. Indian Journal of Microbiology, 53, 321-322.

[61]   Jimenez, J.N., Ocampo, A.M., Vanegas, J.M., Rodriguez, E.A., Garces, C.G., Patino, L.A., Ospina, S. and Correa, M.M. (2011) Characterisation of Virulence Genes in Methicillin Susceptible and Resistant Staphylococcus aureus Isolates from a Paediatric Population in a University Hospital of Medellin, Colombia. Memórias do Instituto Oswaldo Cruz, 106, 980-985.

[62]   Cameron, D.R., Howden, B.P. and Peleg, A.Y. (2011) The Interface between Antibiotic Resistance and Virulence in Staphylococcus aureus and Its Impact upon Clinical Outcomes. Clinical Infectious Diseases, 53, 576-582.

[63]   Pourmand, M.R., Memariani, M., Hoseini, M. and Yazdchi, S.B. (2009) High Prevalence of SEA Gene among Clinical Isolates of Staphylococcus aureus in Tehran. Acta Medica Iranica, 47, 357-361.

[64]   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. Applied and Environmental Microbiology, 66, 1347-1353.

[65]   Vasconcelos, N.G., Pereira, V.C., Araujo Jr., J.P. and da Cunha, M.L. (2011) Molecular Detection of Enterotoxins E, G, H and I in Staphylococcus aureus and Coagulase-Negative Staphylococci Isolated from Clinical Samples of Newborns in Brazil. Journal of Applied Microbiology, 111, 749-762.