High diversity of genetic lineages and virulence genes in nasal Staphylococcus aureus isolates from donkeys destined to food consumption in Tunisia with predominance of the ruminant associated CC133 lineage

BMC Veterinary Research

Carmen Torres¹  Abdellatif Boudabous²  Myriam Zarazaga¹  Naouel Klibi²  Ahlem Jouini²  Carmen Lozano¹  Elena Gómez-Sanz¹  Karim Ben Slama²  Rym Ben Sallem²  Haythem Gharsa² 

[1] Departamento de Agricultura y Alimentación, Área de Bioquímica y Biología Molecular, Universidad de La Rioja, Madre de Dios 51, Logroño, 26006, Spain;Laboratoire des Microorganismes et Biomolécules Actives, Faculté des Sciences de Tunis, Université Tunis-El Manar, Tunis, 2092, Tunisia

关键词: Tunisia;    Virulence genes;    CC133;    spa types;    Donkey;    Staphylococcus aureus;   

Others  :  1119684
DOI  :  10.1186/1746-6148-8-203

 received in 2012-05-31, accepted in 2012-10-22,  发布年份 2012

【 摘 要 】

Background

The objective of this study was to determine the genetic lineages and the incidence of antibiotic resistance and virulence determinants of nasal Staphylococcus aureus isolates of healthy donkeys destined to food consumption in Tunisia.

Results

Nasal swabs of 100 donkeys obtained in a large slaughterhouse in 2010 were inoculated in specific media for S. aureus and methicillin-resistant S. aureus (MRSA) recovery. S. aureus was obtained in 50% of the samples, being all of isolates methicillin-susceptible (MSSA). Genetic lineages, toxin gene profile, and antibiotic resistance mechanisms were determined in recovered isolates. Twenty-five different spa-types were detected among the 50 MSSA with 9 novel spa-types. S. aureus isolates were ascribed to agr type I (37 isolates), III (7), II (4), and IV (2). Sixteen different sequence-types (STs) were revealed by MLST, with seven new ones. STs belonging to clonal clomplex CC133 were majority. The gene tst was detected in 6 isolates and the gene etb in one isolate. Different combinations of enterotoxin, leukocidin and haemolysin genes were identified among S. aureus isolates. The egc-cluster-like and an incomplete egc-cluster-like were detected. Isolates resistant to penicillin, erythromycin, fusidic acid, streptomycin, ciprofloxacin, clindamycin, tetracycline, or chloramphenicol were found and the genes blaZ, erm(A), erm(C), tet(M), fusC were identified.

Conclusions

The nares of donkeys frequently harbor MSSA. They could be reservoirs of the ruminant-associated CC133 lineage and of toxin genes encoding TSST-1 and other virulence traits with potential implications in public health. CC133 seems to have a broader host distribution than expected.

【 授权许可】

   
2012 Gharsa et al.; licensee BioMed Central Ltd.

【 参考文献 】

• [1]Dinges MM, Orwin PM, Schlievert PM: Exotoxins of Staphylococcus aureus. Clin Microbiol Rev 2000, 13:16-34.
• [2]Somerville GA, Proctor RA: At the crossroads of bacterial metabolism and virulence factor synthesis in Staphylococci. Microbiol Mol Biol Rev 2009, 73:233-248.
• [3]Kahl BC, Belling G, Becker , Chatterjee I, Wardecki K, Hilgert K, Cheung AL, Peters G, Herrmann M: Thymidine-dependent Staphylococcus aureus small-colony variants are associated with extensive alterations in regulator and virulence gene expression profiles. Infect Immun 2005, 73:4119-4126.
• [4]Vanderhaeghen W, Hermans K, Haesebrouck F, Butaye P: Methicillin-resistant Staphylococcus aureus (MRSA) in food production animals. Epidemiol Infect 2010, 138:606-625.
• [5]Gómez-Sanz E, Torres C, Lozano C, Fernández-Pérez R, Aspiroz C, Ruiz-Larrea F, Zarazaga M: Detection, molecular characterization, and clonal diversity of methicillin-resistant Staphylococcus aureus CC398 and CC97 in Spanish slaughter pigs of different age groups. Food Pathog Dis 2010, 7:1269-1277.
• [6]Fitzgerald JR: Livestock-associated Staphylococcus aureus: origin, evolution and public health threat. Trends Microbiol 2012, 20:192-198.
• [7]Pantosti A: Methicillin-resistant Staphylococcus aureus associated with animals and its relevance to human health. Frontiers Microbiol 2012, 3:127.
• [8]Weese JS, Duijkeren VE: Methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in veterinary medicine. Vet Microbiol 2009, 140:418-429.
• [9]Burton S, Reid-Smith R, McClure JT, Weese JS: Staphylococcus aureus colonization in healthy horses in Atlantic Canada. Can Vet J 2008, 49:797-799.
• [10]Van den Eede A, Martens A, Lipinska U, Struelens M, Deplano A, Denis O, Haesebrouck F, Gasthuys F, Hermans K: High occurrence of methicillin-resistantStaphylococcus aureusST398 in equine nasal samples. Vet Microbiol 2009, 133:138-144.
• [11]Foti M, Fisichella V, Giacopello C: Detection of methicillin-resistantStaphylococcus aureus (MRSA) in the microbial flora from the conjunctiva of healthy donkeys from Sicily (Italy). Vet Ophthalmol 2012, 1463-5224.
• [12]Gutema DF, Duguma BE, Dinka AG: Isolation and identification of aerobic bacterial flora from the upper respiratory tract of donkeys in Central Ethiopia. Int J Appl Res Vet Med 2009, 7:181-189.
• [13]Pilla R, Dapra V, Zecconi A, Piccinini R: Hygienic and health characteristics of donkey milk during a follow-up study. J Dairy Res 2010, 77:392-397.
• [14]Polidori P, Vincenzetti S, Cavallucci C, Beghelli D: Quality of donkey meat and carcass characteristics. Meat Sci 2008, 80:1222-1224.
• [15]Wilson RT: Distribution and importance of the domestic donkey in circumsaharan Africa. Sing J Trop Geog 1978, 2:136-143.
• [16]Nengomasha EM, Pearson RA, Wold AG: Empowering people through donkey power into the next millennium. Empowering Farmers with Animal Traction (Proceedings of an ATNESA Workshop) 2000, 21:22-31.
• [17]Walther B, Monecke S, Ruscher C, Friedrich AW, Ehricht R, Slickers P, Soba A, Wleklinski CG, Wieler LH, Lubke-Becker A: Comparative molecular analysis substantiates zoonotic potential of equine methicillin-resistant Staphylococcus aureus. J Clin Microbiol 2009, 47:704-710.
• [18]CLSI: Performance standards for antimicrobial susceptibility testing; eighteenth informational supplement. Wayne, Pennsylvania, USA: CLSI document M100-18 CLSI; 2010.
• [19]O’Neill AJ, Bostock M, Moita AM, Chopra I: Antimicrobial activity and mechanisms of resistance to cepahalosporin P1, an antibiotic related to fusidic acid. J Antimicrob Chemother 2002, 50:839-848.
• [20]Harmsen D, Claus H, Witte W, Rothganger J, Claus H, Turnwald D, Vogel U: Typing of methicillin-resistant Staphylococcus aureus in a university hospital setting by using novel software for spa repeat determination and database management. J Clin Microbiol 2003, 41:5442-5448.
• [21]Shopsin B, Herring S, Kreiswirth BN: Hospital-acquired and community-derived: the future of MRSA? Clin Infect Dis 2003, 37:151-152. author reply 152
• [22]Guinane CM, Ben Zakour NL, Tormo-Mas MA, Weinert LA, Lowder BV, Cartwright RA, Smyth DS, Smyth CJ, Lindsay JA, Gould KA, Witney A, Hindsb J, Bollback JP, Rambaut A, Penadés JR, Fitzgerald JR: Evolutionary genomics of Staphylococcus aureus reveals insights into the origin and molecular basis of ruminant host adaptation. Gen Biol Evol 2010, 2:454-466.
• [23]Hwang SY, Kim SH, Jang EJ, Kwon NH, Park YK, Koo HC, Jung WK, Kim JM, Park YH: Novel multiplex PCR for the detection of the Staphylococcus aureus superantigen and its application to raw meat isolates in Korea. Int J Food Microbiol 2007, 117:99-105.
• [24]Jarraud S, Mougel C, Thioulouse J, Lina G, Meugnier H, Forey F, Nesme X, Etienne J, Vandenesch F: Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups (alleles), and human disease. Infect Immun 2002, 70:631-641.
• [25]Grundmann H, Aanensen DM, Van den Wijngaard CC, Spratt BG, Harmsen D, Friedrich AW, European Staphylococcal Reference Laboratory Working Group: Geographic distribution of Staphylococcus aureus causing invasive infections in Europe: a molecular-epidemiological analysis. PLoS Med 2010, 7:1-e1000215.
• [26]Rabello RF, Moreira BM, Lopes RMM, Teixeira LM, Riley LW, Castro ACD: Multilocus sequence typing of Staphylococcus aureus isolates recovered from cows with mastitis in Brazilian dairy herds. J Med Microbiol 2007, 56:1505-1511.
• [27]Hasman H, Moodley A, Guardabassi L, Stegger M, Skov RL, Aarestrup FM: spa type distribution in Staphylococcus aureus originating from pigs, cattle and poultry. Vet Microbiol 2010, 141:326-331.
• [28]Porrero MC, Hasman H, Vela AI, Fernández-Garayzábal JF, Domínguez L, Aarestrup FM: Clonal diversity of Staphylococcus aureus originating from the small ruminants goats and sheep. Vet Microbiol 2012, 156:157-161.
• [29]Gharsa H, Ben Slama K, Lozano C, Gómez-Sanz E, Klibi N, Ben Sallem R, Gómez P, Zarazaga M, Boudabous A, Torres C: Prevalence, antibiotic resistance, virulence traits and genetic lineages of Staphylococcus aureus in healthy sheep in Tunisia. Vet Microbiol 2011, 156:367-373.
• [30]Almeida LM, Almeida MZ, Mendonça CL, Mamizuka EM: Novel sequence types (STs) of Staphylococcus aureus isolates causing clinical and subclinical mastitis in flocks of sheep in the northeast of brazil. J Dairy Res 2011, 78:373-378.
• [31]Monecke S, Kuhnert P, Hotzel H, Slickers P, Ehricht R: Microarray based study on virulence-associated genes and resistance determinants of Staphylococcus aureus isolates from cattle. Vet Microbiol 2007, 125:128-140.
• [32]Espinosa-Gongora C, Chrobak D, Moodley A, Bertelsen MF, Guardabassi L: Occurrence and distribution of Staphylococcus aureus lineages among zoo animals. Vet Microbiol 2012. Article in press
• [33]Sung JML, Lloyd DH, Lindsay JA: Staphylococcus aureus host specificity: comparative genomics of human versus animal isolates by multi-strain microarray. Microbiol 2008, 154:1949-1959.
• [34]Franco A, Hasman H, Iurescia M, Lorenzetti R, Stegger M, Pantosti A, Feltrin F, Ianzano A, Porrero MC, Liap M, Battist A: Molecular characterization of spa type t127, sequence type 1 methicillin-resistant Staphylococcus aureus from pigs. J Antimicrob Chemother 2011, 66:1231-5.
• [35]McDonald M, Dougall A, Holt D, Huygens F, Oppedisano F, Giffard PM, Inman-Bamber J, Stephens AJ, Towers R, Carapetis JR, Currie BJ: Use of a single-nucleotide polymorphism genotyping system to demonstrate the unique epidemiology of methicillin-resistant Staphylococcus aureus in remote aboriginal communities. J Clin Microbiol 2006, 44:3720-3727.
• [36]Collery MM, Smyth DS, Twohig JM, Shore AC, Coleman DC, Smyth CJ: Molecular typing of nasal carriage isolates of Staphylococcus aureus from an Irish university student population based on toxin gene PCR, agr locus types and multiple locus, variable number tandem repeat analysis. J Med Microbiol 2008, 57:348-358.
• [37]Collery MM, Smyth DS, Tumilty JJG, Twohig JM, Smyth CJ: Associations between enterotoxin gene cluster types egc1, egc2 and egc3, agr types, enterotoxin and enterotoxin-like gene profiles, and molecular typing characteristics of human nasal carriage and animal isolates of Staphylococcus aureus. J Med Microbiol 2009, 58:13-25.
• [38]Coombs GW, Monecke S, Pearson JC, Tan H, Chew YK, Wilson L, Ehricht R, O’Brien FG, Christiansen KJ: Evolution and diversity of community-associated methicillin-resistant Staphylococcus aureus in a geographical region. BMC Microbiol 2011, 11:215-227. BioMed Central Full Text
• [39]Argudín MA, Mendoza C, Vazquez A, Guerra B, Rodicio MR: Molecular typing of Staphylococcus aureus blood stream isolates from geriatric patients attending a long-term care Spanish hospital. J Med Microbiol 2011, 60:172-179.
• [40]Argudín MA, Mendoza MC, Méndez FJ, Martín MC, Guerra B, Rodicio MR: Clonal complexes and diversity of exotoxin gene profiles in methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates from patients in a Spanish Hospital. J Clin Microbiol 2009, 47:2097-2105.
• [41]Ghasemzadeh-Moghaddama H, Ghaznavi-Rada E, Sekawia Z, Yun-Khoona L, Azizc MN, Hamata RA, Mellesd DC, Belkumd AV, Shamsudina MN, Neela V: Methicillin-susceptible Staphylococcus aureus from clinical and community sources are genetically diverse. Int J Med Microbiol 2011, 30:347-353.
• [42]Breurec S, Fall1 C, Pouillot R, Boisier P, Brisse S, Diene-Sarr F, Djibo S, Etienne JM, Fonkoua C, Perrier-Gros-Claude JD, Ramarokoto CE, Randrianirina FJ, Thiberge M, Zriouil SB, Garin B, Laurent F: Epidemiology of methicillin susceptible Staphylococcus aureus lineages in five major African towns: high prevalence of Panton–Valentine leukocidin genes. Clin Microbiol Infect 2010, 17:633-639.
• [43]Moodley A, Stegger M, Bagcigil1 AF, Baptiste KE, Loeffler A, Lloyd DH, Williams NJ, Leonard N, Abbott Y, Skov R, Guardabassi L: Spa typing of methicillin-resistant Staphylococcus aureus isolated from domestic animals and veterinary staff in the UK and Ireland. J Antimicrob Chemother 2006, 58:1118-1123.
• [44]Ben Slama K, Gharsa H, Klibi N, Jouini A, Lozano C, Gómez-Sanz E, Zarazaga M, Boudabous A, Torres C: Nasal carriage ofStaphylococcus aureus in healthy humans with different levels of contact with animals in Tunisia: genetic lineages, methicillin resistance, and virulence factors. Eur J Clin Microbiol Infect Dis 2011, 30:499-508.
• [45]Lozano C, Gómez-Sanz E, Benito D, Aspiroz C, Zarazaga M, Torres C: Staphylococcus aureus nasal carriage, virulence traits, antibiotic resistance mechanisms, and genetic lineages in healthy humans in Spain, with detection of CC398 and CC97 strains. Int J Med Microbiol 2011, 301:500-505.
• [46]Smyth DS, Hartigan PJ, Meaney WJ, Fitzgerald JR, Deobald CF, Bohach GA, Smyth CJ: Superantigen genes encoded by the egc cluster and SaPIbov are predominant among Staphylococcus aureus isolates from cows, goats, sheep, rabbits and poultry. J Med Microbiol 2005, 54:401-411.
• [47]Sobral D, Schwarz S, Bergonier D, Brisabois A, Feßler AT, Gilbert FB, Kadlec K, Lebeau B, Loisy-Hamon F, Treilles M, Pourcel C, Vergnaud G: High throughput multiple locus variable number of tandem repeat analysis (MLVA) of Staphylococcus aureus from human, animal and food sources. PLoS One 2012, 7:e33967.
• [48]Chen HJ, Hung WC, Tseng SP, Tsai JC, Hsueh PR, Teng LJ: Fusidic acid resistance determinants in Staphylococcus aureus clinical isolates. Antimicrob Agents Chemother 2010, 54:4985-4991.