Deteksi Hemaglutinin, Hemolisin dan Koagulase Secara Fenotipik dan Genotipik pada Staphylococcus aureus Isolat Asal Broiler

https://doi.org/10.22146/jsv.38434

Khusnan Khusnan(1*), Dwi Kusmanto(2)

(1) Akademi Peternakan Brahmaputra, Yogyakarta
(2) Akademi Peternakan Brahmaputra, Yogyakarta
(*) Corresponding Author

Abstract


Staphylococcus aureus is a bacterium causing diseases in animals and human. Staphylococcus aureus in broilers cause septicemia, tendosinovitis, dermatitis, endocarditis, wound infections, arthritis and bumblefoot. In this research, 24 isolates of Staphylococcus aureus from broiler were characterized of its virulent factors including the presence of haemagglutinin, ability to agulate the plasma in tubular coagulase test as well presipitate formation in clumping factor test, and haemolysis types. By polymerase chain reaction (PCR) were genotypically detected genes coa, clf, hlaA, and hlaB. All of the isolates (100%) had haemagglutinin, capable to agglutinate and precipitae of rabbit plasma. All isolates could lyse sheep red blood cells with the type of α-hemolysis (45.8%),  β-hemolysis (50.0%) and γ-haemolysis (4.2%). Genotypically, all isolates (100%) had coa and clf genes,  hlaA gene 7(0.8%) and hlaB gene (29.2%).


Keywords


Staphylococcus aureus; Broiler; Coagulase; Hemolysin; Haemagglutinin

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References

Aarestrup, F.M., Dangler, C.A. and Sordillo, L.M. (1995) Prevalence of coagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastitis. Can. J. Vet. Res., 59:124-128.

Abdalrahman, L.S., Stanley, A., Wells, H. and Fakhr, M.K. (2015) Isolation, virulence, and antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin sensitive Staphylococcus aureus (MSSA) strains from Oklahoma retail poultry meats. Int. J. Environ. Res. Public Health. 12: 6148-6161.

Abrar, M., Wibawan, I.W.T., Priosoeryanto, B.P., Soedarwanto, M. and Pasaribu. F.H. (2013) Role of Staphylococcus aureus haemaglutinin in adhesion process on udder epithelial cells. J. Kedokteran Hewan, 7(1): 43-46.

Akineden, O., Annemuller, C., Hassan, A.A., Laemmler, C., Wolter, W. and 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.

Almeida, L.M., de Almeida, M.Z., de Mendonca, C.L. and Mamizuka, E.M. (2013) Comparative analysis of agr groups and virulence genes among subclinical and clinical mastitis Staphylococcus aureus isolates from sheep flocks of the Northeast of Brazil. Brazilian Journal of Microbiology . 44(2):493-498.

Ariyanti, D., Salasia, S.I.O. and Tato, S. (2011) Characterization of haemolysin of Staphylococcus aureus isolated from food of animal Origin. Indon. J. of Biotech., 16(1): 32-37.

Aslantas, O., Demir, C., Turutoglu, H., Cantekin, S., Ergun, Y. and Dogruer, G. (2007) Coagulase gene polymorphism of Staphylococcus aureus isolated form sub clinical mastitis. Turk. J. Vet. Anim. Sci. 31:253-257

Aziza, A.M.S., Akter, M.R., Rahaman, M.S., Mas, K., Zakm, H., Jahan, M.S. and Kabir, L.S.M. (2013) Isolation, identification and antibiogram profiles of Staphylococcus aureus from commercial broiler flocks in Dinajpur District of Bangladesh with special focuson the determination of lethal effect of extracted toxin. Sci. J. of Microbiol. 2(4): 74-82.

Baba, T., Takeuchi, F., Kuroda, M., Yuzawa, H., Aoki, K., Oguchi, A., Nagai, Y., Iwama, N., Asano, K., Naimi, T., Kuroda, H., Cui, L., Yamamoto, K. and Hiramatsu, K. (2002) Genome and virulence determinants of high virulence community-acquired MRSA. Lancet, 359: 1819-1827

Babu, N.G.R., Shree G.B.B., Rekha L. and Karthiga P. (2014) Molecular analysis of coagulase (coa) gene polymorphism in clinical isolates of Staphylococcus aureus by PCR-RFLP. Int. J. of Innov. Res. in Sci., Eng. and Technol., 8162-8168.

Bhakdi, S., Muhly, M., Korom, S. and Hugo, F. (1989) Release of interleukin-1 associated with potent cytocidal action of staphylococcal alpha-toxin on human monocytes. Infect. Immun., 57: 3512-3519.

Boerema, J.A, Clemens, R. and Brightwell, G. (2006) Evaluation of molecular methods to determine enterotoxigenic status and molecular genotype of bovine, ovine, human and food isolates of Staphylococcus aureus. Int. J. Food Microbiol., 107: 192-201.

Bronner, S., Monteil, H. and Prevost, G. (2004) Regulation of virulence determinants in Staphylococcus aureus: complexity and applications. FEMS Microbiol. Rev., 28(2): 183-200.

Brückler, J., Schwarz, S. and Untermann, F. (1994) Staphylokokken-Infektionen und–Enterotoxine, Band. II/1, In Blobel, H. und Schlieber (eds.), Handbuch der bakteriellen Infektionen bei Tieren, 2. Auflage. Gustav Fischer Verlag Jena, Stuttgart.

Burnside, K., Lembo, A., de los Reyes, M., Iliuk, A., Nguyen-Thao, B., Connelly, J.E., Wan-Jung, L., Schmidt, B.Z., Richardson, A.R., Fang, F.C., Tao, W.A. and Rajagopal, L. (2010) Regulation of hemolysin expression and virulence of Staphylococcus aureus by a serine/threonine kinase and phosphatase. http://dx.doi.org/10.1371/journal.pone.0011071

Cifrian, E., Guidry, A.J., Bramley, A.J., Norcross, N.L., Bastida-Corcuera, F.D. and Marquardt, W.W. (1996) Effect of staphylococcal alpha toxins on the cytotoxicity, proliferation and adherence of Staphylococcus aureus to bovine mammary epithelial cells. Vet. Microbiol. 48:187-198.

Coelho, S.M.O., Pereira, I.A., Soares, L.C. and Pribul, B.R., and Souza, M.M.S. (2011) Short communication: Profile of virulence factors of Staphylococcus aureus isolated from subclinical bovine mastitis in the state of Rio de Janeiro, Brazil . J. Dairy Sci., 94: 3305-3310.

Cotar, A.I., Chifiriuc, M.C., Dinu, S., Bucur, M., Iordache, C., Banu, O., Dracea, O., Larionand, C. and Lazar, V. (2010) Screening of molecular virulence markers in Staphylococcus aureus and Pseudomonas aeruginosa strains isolated from clinical infections. Int. J. Mol. Sci., 11(12): 5273-5291.

Da Silva, R.E., Boechat, J.U.D., Martins, J.C.D., Ferreira, W.P.B., Sequera, A.P.S. and Da silva N. (2005) Hemolysin production by Staphylococcus aureus species isolated from mastitic goat milk in Brazilian Dairy Herd. Sci. Small rum. res., 56: 271-275.

de los Santos, Fernández, M., Carro, S. and Zunino, P. (2014) Characterisation of Staphylococcus aureus isolated from cases of bovine subclinical mastitis in two Uruguayan dairy farms. Arch. Med. Vet., 46: 315-320.

Delgado, S., Garcia P., Fernandez, L., Jimenez, E., Rodriguez-Banos, M., Del Campo, R. and Jrodriguez, J.M. (2011) Characterization of Staphylococcus aureus strains involved in human and bovine mastitis. Immunol. Med. Microbiol., 62: 225-235.

Diep, B.A. and Otto, M. (2008). The role of virulence determinants in community-associated MRSA pathogenesis. Trends in Microbiol., 16(8): 361-369.

Dinges, M.M., Orwin, P.M. and Schlievert, P.M. (2000) Exotoxins of Staphylococcus aureus. Clin. Microbiol. Rev., 13: 16-34.

El-Hamid, M.I.A. and Bendary, M.M. (2013) Association between agr alleles and toxin gene profiles of S. aureus isolates from human and animals sources in Egypt. Int. J. of Adv. Res., 1(8): 133-144.

Elsayed, M,S., El-Bagoury, A.M. and Dawoud, M.A. (2015) Phenotypic and genotypic detection of virulence factors of Staphylococcus aureus isolated from clinical and subclinical mastitis in cattle and water buffaloes from different farms of Sadat City in Egypt, Vet. World, 8(9): 1051-1058.

Engleberg, N.C., DiRita, V. and Dermody, T.S. (2006) Schaechter’s Mechanisms of Microbial Disease. Fourth edition. Chapter 11: 147-155.

Fei, W., Hongjun, Y., Hong-bin, H., Changfa, W., Yundong, G., Qifeng, Z., Xiaohong, W. and Yanjun, Z. (2011) Study on the haemolysin phenotype and the genotype distribution of Staphylococcus aureus caused bovine mastitis in Shandog dairy farms. Int. J. Appl. Res. Vet. M., 9(4): 416-421.

Ferens, W.A., Davis, W.C., Hamilton, M.J., Park, Y.H., Deobald, C.F. and Fox, L. (1998) Activation of bovine lymphocyte subpopulations by staphylococcal enterotoxin C. Infect. Immun. 66: 573-580.

Fiaschi, L., Di Palo, B., Scarselli, M., Pozzi, C., Tomaszewski, K., Galletti, B., Nardi- Dei, V., Arcidiacono, L., Mishra, R.P.N., Mori, E., Pallaoro, M., Falugi, F., Torre, A., Fontana, M.R., Soriani, M., Wardenburg, J.B., Grandi, G., Rappuoli, R., Ferlenghi, I. and Bagnoli, F. (2016) Auto-assembling detoxified Staphylococcus aureus alpha-hemolysin mimicking the wild-type cytolytic toxin. Clin. Vaccine Immunol., 23: 442-450.

Flick, M.J., Du, X., Prasad, J.M., Raghu, H., Palumbo, J.S., Smeds, E., Hook, M. and Degen, J.L. (2013) Genetic elimination of the binding motif on fibrinogen for the S. aureus virulence factor clfA improves host survival in septicemia. Blood. 121: 1783-1794.

Foster, T.J. and Hook, M. (1998) Surface protein adhesins of Staphylococcus aureus. Trends Microbiol. 6: 484-488

Franco, G.J.C., Gonzalez, V.L.G., Gomez, M.S.C., Carrillo, G.J.M. and Ramirez, C.J.J. (2008) Virulence factors analysis of Staphylococcus aureus isolated from bovine mastitis in Mexico. e­Gnosis [online] 6, Art. 7. http://www.redalyc.org/pdf/730/73011197007.pdf

Gatermann, S.G., Kreft, B., Marre, R. and Wanner, G. (1992) Identification and characterization of a surface-associated protein (ssp) of Staphylococcus saprophyticus. Infect. Immun. 60: 1055-1060.

Gordon, R. and Lowy, F.C. (2008) Pathogenesis of methicillin-resistant Staphylococcus aureus infection. Clin. Infect. Dis. 46: 350-359.

Guggenberger, C., Wolz, C., Morrissey, J.A., and Heesemann, J. (2012) Two distinct coagulase-dependent barriers protect Staphylococcus aureus from neutrophils in a three dimensional in vitro infection model. PLoS Pathog., 8: e1002434 2012

Harshini, A.K. (2015) Adhesion in Bacteria. Int J. of Sci and res., 279-281

Haveri, M., Hovinen, M., Roslof, A., Pyorala, S. (2008) Molecular types and genetic profiles of Staphylococcus aureus strains isolated from bovine intramammary infections and extramammary sites. J. Clin. Microbiol., 46: 3728-3735.

Hayashida, A., Bartlett A.H., Foster T.J. and Park P.W. (2009) Staphylococcus aureus beta-toxin induces lung injury through syndecan-1. Am. J. Pathol. 174: 509-518.

Higgins J., Loughman, A., van Kessel, K.P.M., van Strijp, A.G. and Timothy, J. (2006) Foster clumping factor A of Staphylococcus aureus inhibits phagocytosis by human polymorphonuclear leucocytes. FEMS. Microbiol. Lett., 258: 290-297

Karakulska, J., Pobucewicz, A., Nawrotek, A., Muszyńska, M., Furowicz, A.J. and Furowicz D.C. (2011) Molecular typing of Staphylococcus aureus based on PCR-RFLP of coa gene and RAPD analysis. Pol. J. Vet. Sci., 14: 285-286.

Khusnan dan Kusmanto, D. (2014) Methicillin-Resistant Staphylococcus aureus (MRSA) Isolat ayam pedaging dan kajian virulensi pada sel hospes secara invivo dan invitro. Laporan Penelitian Fundamental. Kemendikbud. Republik Indonesia

Kolar, S.L., Ibarra, J.A., Rivera, F.E., Mootz, J.M., Davenport, J.E., Stevens, S.M., Horswill, A.R. and Shaw, L.N. (2013) Extracellular proteases are key mediators of Staphylococcus aureus virulence via the global modulation of virulence determinant stability. Microbiol. Open., 2:18-34.

Kumar, R., Yadav, B.R. and Singh, R.S. (2011) Antibiotic resistance and pathogenicity factors in Staphylococcus aureus isolated from mastitic sahiwal cattle. J. Biosci., 36(1):175-188.

Larsen, H.D., Aarestrup, F.M. and Jensen, N.E. (2002) Geographical variation in the presence of genes encoding superantigenic exotoxins and beta-hemolysin among Staphylococcus aureus isolated from bovine mastitis in Europe and USA. Vet. Microbiol., 85: 61-67.

Leitner, G., Krifucks, O., Glickman, A., Younis, A. and Saran, A. (2003) Staphylococcus aureus strains isolated from bovine mastitis: virulence, antibody production and protection from challenge in a mouse model. Immunol. and Med. Microbiol., 35: 99-106.

Marques, V.F., De Souza, M.M.S., De Mendonça, E.C.L., De Alencar, T.A., Pribu, B.R., De Coelho, S., Lasagno, M. and Reinoso, E.B. (2013) Phenotypic and genotypic analysis of virulence in Staphylococcus spp. and its clonal dispersion as a contribution to the study of bovine mastitis. Pesq. Vet. Bras., 33(2): 161-170.

McAdow, M., Missiakas D.M. and Schneewind, O. (2012) Staphylococcus aureus secretes coagulase and von willebrand factor binding protein to modify the coagulation cascade and establish host infections. J. Innate. Immun., 4: 141-148

McNamee, P.T. and Smyth, J.A. (2000) Bacterial chondronecrosis with osteomylitis (femoral head necrosis) of broiler chickens: a review. Av. Pathol. 29(5): 477-495.

Memon, J., Yang, Y., Kashifa, J., Yaqoob, M., Buriroa, R., Soomroa, J., Liping, W. and Hongjie, F. (2013) Genotypes, virulence factors and antimicrobial resistance genes of Staphylococcus aureus isolated in bovine subclinical mastitis from Eastern China. Pak. Vet. J., 33(4): 486-491.

Microbeonline (2013) Blood Agar: Composition, preparation, uses and types of hemolysis. http://microbeonline.com/blood-agar-composition-preparation-uses-and-types-of-hemolysis/

Mohajeri, P., Azizkhani, S., Farahani, A. and Norozi, B. (2016) Genotyping of coa and aroA Genes of Methicillin-Resistant Staphylococcus aureus Strains Isolated From Nasal Samples in Western Iran. Jundishapur J. Microbiol., 9(1): e26460

Mohamed, S.R., Habashy, H.F., El-Shahawy, H.S. and Salem, N.I.E. (2017) Phenotype and genotype characterization of staphylococcus aureus causing sub clinical mastitic cattle milk using vitek2 compact system and PCR. An. Health Res. J. 5(1): 86-96.

Mohan, K., Shroeder-Tucker, L.C., Karenga, D., Dziva, F., Harrison, A. and Muvavarirwa, P. (2002) Unidentified coryneform bacterial strain from cases of polyarthritis in chickens: phenotype and fatty acid profile. Av. Dis., 46: 1051-1054.

Moraveji, Z., Tabatabaei, M., Aski, H.S. and Khoshbakht, R. (2014) Characterization of hemolysins of Staphylococcus strains isolated from human and bovine, southern Iran. IJVR, 15(4): 326-330.

Moreillon, P., Entenza, J.M., Francioli, P., Mcdevitt, D., Foster, T.J., Francois, P. and Vaudaux, P. (1995) Role of Staphylococcus aureus coagulase and clumping factor in pathogenesis of experimental endocarditis. Am. Soc. for Microbiol. Infect. and Immun., 63: 4738-4743.

Park, P.W., Foster, T.J., Nishi, E., Duncan, S.J., Klagsbrun, M. and Chen, Y. (2004) Activation of syndecan-1 ectodomain shedding by Staphylococcus aureus α-toxin and β-toxin. J. Biol. Chem. 279(1): 251-258.

Peacock, S.J., de Silva, I. and Lowy, F.D. (2002) What determines nasal carriage of Staphylococcus aureus? Trends Microbiol., 9: 605-610.

Perkins, S., Walsh, E.J., Deivanayagam, C.C., Narayana, S.V., Foster, T.J. and Hook, M. (2001) Structural organization of the fibrinogen-binding region of the clumping factor B MSCRAMM of Staphylococcus aureus. J. Biol. Chem., 276: 44721-44728.

Rares, F.E.S. (2011) Deteksi hemaglutinin protein permukaan Staphylococcus aureus dengan menggunakan eritrosit Rattus novergicus strain wistar. J. Biomedik, 3(3): 156-164. (abstract)

Rasheed, B.Y. (2011) Isolation and identification of bacteria causing arthritis in chickens Iraqi J. of Vet. Sci., 25(2): 93-95.

Rupp, M.E. and Archer, G.L. (1995) Hemagglutination and adherence to plastic by Staphylococcus epidermidis. Infect. Immun., 60: 4322-432.

Rupp, M.E., Sloot, N., Meyer, H.G., Han, J. and Gatermann, S. (1995) Characterization of the hemagglutinin of Staphylococcus epidermidis. J. Infect. Dis., 172: 1509-1518

Rusenova, N.V. and Rusenov, A.G. (2017) Detection of Staphylococcus aureus among coagulase positive staphylococci from animal origin based on conventional and molecular methods. Mac. Vet. Rev., 40(1): 29-36.

Salasia, S.I.O. and Laemmler, C. (1995) Occurrence of haemagglutinating adhesin among virulent and avirulent isolates of Streptococcus suis. Med. Sci. Res. 22:763-764

Salasia, S.I.O., Tato, S., Sugiyono, N., Ariyanti, D. and Prabawati, F. (2011) Genotypic characterization of Staphylococcus aureus isolated from bovines, humans, and food in Indonesia. J. Vet. Sci., 12(4): 353-361

Salasia, S.I.O., Khusnan, Z., Laemmler, C. and Zschock, M. (2004) Comparative studies on phenotypic and genotypic properties of Staphylococcus aureus isolated from bovine subclinical mastitis in central Java in Indonesia and Hesse in Germany. J. Vet. Sci., 5(2): 103-109.

Singh R., Ray P., Das A. and Sharma M. (2009) Role of persisters and small-colony variants in antibiotic resistance of planktonic and biofilm-associated Staphylococcus aureus: an in vitro study. J. Med. Microbiol., 58: 1067-1073.

Tirpude, R.J. and Batra, H.V. (2012) Characteristics of Staphylococcus aureus isolated from acute, sub-acute and sub-clinical staphylococcosis in rabbits. World Rabbit Sci., 20: 215-221.

Tiwari, H.K., Sapkota, D. and Sen, M.R. (2008) Evaluation of different tests for detection of Staphylococcus aureus using coagulase (coa) gene PCR as the gold standard. Nepal Med. Coll. J., 10(2): 129-131.

Vancraeynest, D., Haesebrouck, F., Deplano, A., Denis, O., Godard, C., Wildemauwe, C. and K. Hermans K. (2006) International Dissemination of a High Virulence Rabbit Staphylococcus aureus Clone. J. Vet. Med., 53(9): 418-422.

Vaudaux, P.E., François, P., Proctor, R.A., McDevitt, D., Foster, T.J., Albrecht, R.M, Lew, D.P., Wabers, H. and Cooper, S.L. (1995) Use of adhesion-defective mutants of Staphylococcus aureus to define the role of specific plasma proteins in promoting bacterial adhesion to canine arteriovenous shunts. Infect. Immun., 63(2): 585-90.

Vanderhaeghen, W., Hermans, K., Haesebrouck, F. and Butaye, P. (2010) Methicillin-resistant Staphylococcus aureus (MRSA) in food production animals. Epidemiol. Infect., 138: 606-625. .

Wibawan, I.W.T., Laemmler, C. and Pasaribu, F.H. (1992) A hemaglutining adesion of group B Streptococci isolates from cases of bovine mastitis mediated adherence to Hela Cell. J. Gen Microbiol., 139: 2173-1242



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