In the early 2000s, a particular MRSA clonal complex (CC398) was found mainly in pigs and pig farmers in Europe. Since then, CC398 has been detected among a wide variety of animal species worldwide. We investigated the population structure of CC398 through mutation discovery at 97 genetic housekeeping loci, which are distributed along the CC398 chromosome within 195 CC398 isolates, collected from various countries and host species, including humans. Most of the isolates in this collection were received from collaborating microbiologists, who had preserved them over years. We discovered 96 bi-allelic polymorphisms, and phylogenetic analyses revealed that an epidemic sub-clone within CC398 (dubbed ‘clade (C)’) has spread within and between equine hospitals, where it causes nosocomial infections in horses and colonises the personnel. While clade (C) was strongly associated with S. aureus from horses in veterinary-care settings (p = 2×10?7), it remained extremely rare among S. aureus isolates from human infections.
References
[1]
Lowy FD (1998) Staphylococcus aureus infections. N Engl J Med 339: 520–532. doi: 10.1056/nejm199808203390806
[2]
Devriese LA, Van Damme LR, Fameree L (1972) Methicillin (cloxacillin)-resistant Staphylococcus aureus strains isolated from bovine mastitis cases. Zentralbl Veterinarmed B 19: 598–605. doi: 10.1111/j.1439-0450.1972.tb00439.x
[3]
Scott GM, Thomson R, Malone-Lee J, Ridgway GL (1988) Cross-infection between animals and man: possible feline transmission of Staphylococcus aureus infection in humans? J Hosp Infect 12: 29–34. doi: 10.1016/0195-6701(88)90119-3
[4]
Cefai C, Ashurst S, Owens C (1994) Human carriage of methicillin-resistant Staphylococcus aureus linked with pet dog. Lancet 344: 539–540. doi: 10.1016/s0140-6736(94)91926-7
[5]
Voss A, Loeffen F, Bakker J, Klaassen C, Wulf M (2005) Methicillin-resistant Staphylococcus aureus in pig farming. Emerg Infect Dis 11: 1965–1966. doi: 10.3201/eid1112.050428
[6]
Witte W, Strommenger B, Stanek C, Cuny C (2007) Methicillin-resistant Staphylococcus aureus ST398 in humans and animals, Central Europe. Emerg Infect Dis 13: 255–258. doi: 10.3201/eid1302.060924
[7]
Guardabassi L, Stegger M, Skov R (2007) Retrospective detection of methicillin resistant and susceptible Staphylococcus aureus ST398 in Danish slaughter pigs. Vet Microbiol 122: 384–386. doi: 10.1016/j.vetmic.2007.03.021
[8]
Smith TC, Male MJ, Harper AL, Kroeger JS, Tinkler GP, et al. (2009) Methicillin-resistant Staphylococcus aureus (MRSA) strain ST398 is present in midwestern U.S. swine and swine workers. PLoS One 4: e4258. doi: 10.1371/journal.pone.0004258
[9]
Khanna T, Friendship R, Dewey C, Weese JS (2008) Methicillin resistant Staphylococcus aureus colonization in pigs and pig farmers. Vet Microbiol 128: 298–303. doi: 10.1016/j.vetmic.2007.10.006
[10]
Pan A, Battisti A, Zoncada A, Bernieri F, Boldini M, et al. (2009) Community-acquired methicillin-resistant Staphylococcus aureus ST398 infection, Italy. Emerg Infect Dis 15: 845–847. doi: 10.3201/eid1505.081417
[11]
Agerso Y, Hasman H, Cavaco LM, Pedersen K, Aarestrup FM (2012) Study of methicillin resistant Staphylococcus aureus (MRSA) in Danish pigs at slaughter and in imported retail meat reveals a novel MRSA type in slaughter pigs. Vet Microbiol 157: 246–250. doi: 10.1016/j.vetmic.2011.12.023
[12]
Nemati M, Hermans K, Lipinska U, Denis O, Deplano A, et al. (2008) Antimicrobial resistance of old and recent Staphylococcus aureus isolates from poultry: first detection of livestock-associated methicillin-resistant strain ST398. Antimicrob Agents Chemother 52: 3817–3819. doi: 10.1128/aac.00613-08
[13]
Cuny C, Friedrich A, Kozytska S, Layer F, Nübel U, et al. (2010) Emergence of methicillin-resistant Staphylococcus aureus (MRSA) in different animal species. Int J Med Microbiol 300: 109–117. doi: 10.1016/j.ijmm.2009.11.002
[14]
Kehrenberg C, Cuny C, Strommenger B, Schwarz S, Witte W (2009) Methicillin-resistant and -susceptible Staphylococcus aureus strains of clonal lineages ST398 and ST9 from swine carry the multidrug resistance gene cfr. Antimicrob Agents Chemother 53: 779–781. doi: 10.1128/aac.01376-08
[15]
Tavakol M, Olde Riekerink RG, Sampimon OC, Van Wamel WJ, Van Belkum A, et al. (2012) Bovine-associated MRSA ST398 in the Netherlands. Acta Vet Scand 54: 28. doi: 10.1186/1751-0147-54-28
[16]
Monecke S, Kuhnert P, Hotzel H, Slickers P, Ehricht R (2007) Microarray based study on virulence-associated genes and resistance determinants of Staphylococcus aureus isolates from cattle. Vet Microbiol 125: 128–140. doi: 10.1016/j.vetmic.2007.05.016
[17]
Persoons D, Van Hoorebeke S, Hermans K, Butaye P, de Kruif A, et al. (2009) Methicillin-resistant Staphylococcus aureus in poultry. Emerg Infect Dis 15: 452–453. doi: 10.3201/eid1503.080696
[18]
Fessler AT, Olde Riekerink RG, Rothkamp A, Kadlec K, Sampimon OC, et al. (2012) Characterization of methicillin-resistant Staphylococcus aureus CC398 obtained from humans and animals on dairy farms. Vet Microbiol.
[19]
Fessler A, Scott C, Kadlec K, Ehricht R, Monecke S, et al. (2010) Characterization of methicillin-resistant Staphylococcus aureus ST398 from cases of bovine mastitis. J Antimicrob Chemother 65: 619–625. doi: 10.1093/jac/dkq021
[20]
Fessler AT, Kadlec K, Hassel M, Hauschild T, Eidam C, et al. (2011) Characterization of methicillin-resistant Staphylococcus aureus isolates from food and food products of poultry origin in Germany. Appl Environ Microbiol 77: 7151–7157. doi: 10.1128/aem.00561-11
[21]
Cuny C, Nathaus R, Layer F, Strommenger B, Altmann D, et al. (2009) Nasal colonization of humans with methicillin-resistant Staphylococcus aureus (MRSA) CC398 with and without exposure to pigs. PLoS One 4: e6800. doi: 10.1371/journal.pone.0006800
[22]
van Belkum A, Melles DC, Peeters JK, van Leeuwen WB, van Duijkeren E, et al. (2008) Methicillin-resistant and -susceptible Staphylococcus aureus sequence type 398 in pigs and humans. Emerg Infect Dis 14: 479–483. doi: 10.3201/eid1403.070760
[23]
Graveland H, Duim B, van Duijkeren E, Heederik D, Wagenaar JA (2011) Livestock-associated methicillin-resistant Staphylococcus aureus in animals and humans. Int J Med Microbiol 301: 630–634. doi: 10.1016/j.ijmm.2011.09.004
[24]
van Loo I, Huijsdens X, Tiemersma E, de Neeling A, van de Sande-Bruinsma N, et al. (2007) Emergence of methicillin-resistant Staphylococcus aureus of animal origin in humans. Emerg Infect Dis 13: 1834–1839. doi: 10.3201/eid1312.070384
[25]
Van den Broek, Van Cleef BA, Haenen A, Broens EM, Van der Wolf PJ, et al. (2009) Methicillin-resistant Staphylococcus aureus in people living and working in pig farms. Epidemiol Infect 137: 700–708. doi: 10.1017/s0950268808001507
[26]
Uhlemann AC, Porcella SF, Trivedi S, Sullivan SB, Hafer C, et al. (2012) Identification of a highly transmissible animal-independent Staphylococcus aureus ST398 clone with distinct genomic and cell adhesion properties. MBio 3.
[27]
van der Mee-Marquet N, Francois P, Domelier-Valentin AS, Coulomb F, Decreux C, et al. (2011) Emergence of unusual bloodstream infections associated with pig-borne-like Staphylococcus aureus ST398 in France. Clin Infect Dis 52: 152–153. doi: 10.1093/cid/ciq053
[28]
Price LB, Stegger M, Hasman H, Aziz M, Larsen J, et al. (2012) Staphylococcus aureus CC398: host adaptation and emergence of methicillin resistance in livestock. MBio 3.
[29]
van de Giessen AW, van Santen-Verheuvel MG, Hengeveld PD, Bosch T, Broens EM, et al. (2009) Occurrence of methicillin-resistant Staphylococcus aureus in rats living on pig farms. Prev Vet Med 91: 270–273. doi: 10.1016/j.prevetmed.2009.05.016
[30]
Vincze S, Stamm I, Monecke S, Kopp PA, Semmler T, et al. (2013) Molecular analysis of human and canine Staphylococcus aureus strains reveals distinct extended-host-spectrum genotypes independent of their methicillin resistance. Appl Environ Microbiol 79: 655–662. doi: 10.1128/aem.02704-12
[31]
Cuny C, Strommenger B, Witte W, Stanek C (2008) Clusters of infections in horses with MRSA ST1, ST254, and ST398 in a veterinary hospital. Microb Drug Resist 14: 307–310. doi: 10.1089/mdr.2008.0845
[32]
Van den Eede A, Martens A, Lipinska U, Struelens M, Deplano A, et al. (2009) High occurrence of methicillin-resistant Staphylococcus aureus ST398 in equine nasal samples. Vet Microbiol 133: 138–144. doi: 10.1016/j.vetmic.2008.06.021
[33]
Loeffler A, Lloyd DH (2010) Companion animals: a reservoir for methicillin-resistant Staphylococcus aureus in the community? Epidemiol Infect 138: 595–605. doi: 10.1017/s0950268809991476
[34]
Loeffler A, Kearns AM, Ellington MJ, Smith LJ, Unt VE, et al. (2009) First isolation of MRSA ST398 from UK animals: a new challenge for infection control teams? J Hosp Infect 72: 269–271. doi: 10.1016/j.jhin.2009.04.002
[35]
van Duijkeren E, Moleman M, Sloet van Oldruitenborgh-Oosterbaan MM, Multem J, Troelstra A, et al. (2010) Methicillin-resistant Staphylococcus aureus in horses and horse personnel: an investigation of several outbreaks. Vet Microbiol 141: 96–102. doi: 10.1016/j.vetmic.2009.08.009
[36]
Sieber S, Gerber V, Jandova V, Rossano A, Evison JM, et al. (2011) Evolution of multidrug-resistant Staphylococcus aureus infections in horses and colonized personnel in an equine clinic between 2005 and 2010. Microb Drug Resist 17: 471–478. doi: 10.1089/mdr.2010.0188
[37]
O'Mahony R, Abbott Y, Leonard FC, Markey BK, Quinn PJ, et al. (2005) Methicillin-resistant Staphylococcus aureus (MRSA) isolated from animals and veterinary personnel in Ireland. Vet Microbiol 109: 285–296. doi: 10.1016/j.vetmic.2005.06.003
[38]
Seguin JC, Walker RD, Caron JP, Kloos WE, George CG, et al. (1999) Methicillin-resistant Staphylococcus aureus outbreak in a veterinary teaching hospital: potential human-to-animal transmission. J Clin Microbiol 37: 1459–1463.
Weese JS (2005) Methicillin-resistant Staphylococcus aureus: an emerging pathogen in small animals. J Am Anim Hosp Assoc 41: 150–157.
[41]
Nübel U, Dordel J, Kurt K, Strommenger B, Westh H, et al. (2010) A timescale for evolution, population expansion, and spatial spread of an emerging clone of methicillin-resistant Staphylococcus aureus. PLoS Pathog 6: e1000855. doi: 10.1371/journal.ppat.1000855
[42]
Harris SR, Feil EJ, Holden MT, Quail MA, Nickerson EK, et al. (2010) Evolution of MRSA during hospital transmission and intercontinental spread. Science 327: 469–474. doi: 10.1126/science.1182395
[43]
Parker J, Rambaut A, Pybus OG (2008) Correlating viral phenotypes with phylogeny: accounting for phylogenetic uncertainty. Infect Genet Evol 8: 239–246. doi: 10.1016/j.meegid.2007.08.001
[44]
Nübel U, Roumagnac P, Feldkamp M, Song JH, Ko KS, et al. (2008) Frequent emergence and limited geographic dispersal of methicillin-resistant Staphylococcus aureus. Proc Natl Acad Sci U S A 105: 14130–14135. doi: 10.1073/pnas.0804178105
[45]
Robinson DA, Enright MC (2004) Evolution of Staphylococcus aureus by large chromosomal replacements. J Bacteriol 186: 1060–1064. doi: 10.1128/jb.186.4.1060-1064.2004
[46]
Lowder BV, Guinane CM, Ben Zakour NL, Weinert LA, Conway-Morris A, et al. (2009) Recent human-to-poultry host jump, adaptation, and pandemic spread of Staphylococcus aureus. Proc Natl Acad Sci U S A 106: 19545–19550. doi: 10.1073/pnas.0909285106
[47]
Holden MT, Hsu LY, Kurt K, Weinert LA, Mather AE, et al. (2013) A genomic portrait of the emergence, evolution, and global spread of a methicillin-resistant Staphylococcus aureus pandemic. Genome Res 23: 653–664. doi: 10.1101/gr.147710.112
[48]
Ellington MJ, Hope R, Livermore DM, Kearns AM, Henderson K, et al. (2010) Decline of EMRSA-16 amongst methicillin-resistant Staphylococcus aureus causing bacteraemias in the UK between 2001 and 2007. J Antimicrob Chemother 65: 446–448. doi: 10.1093/jac/dkp448
[49]
Amorim ML, Faria NA, Oliveira DC, Vasconcelos C, Cabeda JC, et al. (2007) Changes in the clonal nature and antibiotic resistance profiles of methicillin-resistant Staphylococcus aureus isolates associated with spread of the EMRSA-15 clone in a tertiary care Portuguese hospital. J Clin Microbiol 45: 2881–2888. doi: 10.1128/jcm.00603-07
[50]
Nulens E, Gould I, MacKenzie F, Deplano A, Cookson B, et al. (2005) Staphylococcus aureus carriage among participants at the 13th European Congress of Clinical Microbiology and Infectious Diseases. Eur J Clin Microbiol Infect Dis 24: 145–148. doi: 10.1007/s10096-004-1258-6
[51]
Wulf MW, Sorum M, van Nes A, Skov R, Melchers WJ, et al. (2008) Prevalence of methicillin-resistant Staphylococcus aureus among veterinarians: an international study. Clin Microbiol Infect 14: 29–34. doi: 10.1111/j.1469-0691.2007.01873.x
[52]
Wulf M, van Nes A, Eikelenboom-Boskamp A, de Vries J, Melchers W, et al. (2006) Methicillin-resistant Staphylococcus aureus in veterinary doctors and students, the Netherlands. Emerg Infect Dis 12: 1939–1941. doi: 10.3201/eid1212.060355
[53]
Hanselman BA, Kruth SA, Rousseau J, Low DE, Willey BM, et al. (2006) Methicillin-resistant Staphylococcus aureus colonization in veterinary personnel. Emerg Infect Dis 12: 1933–1938. doi: 10.3201/eid1212.060231
[54]
Moodley A, Nightingale EC, Stegger M, Nielsen SS, Skov RL, et al. (2008) High risk for nasal carriage of methicillin-resistant Staphylococcus aureus among Danish veterinary practitioners. Scand J Work Environ Health 34: 151–157. doi: 10.5271/sjweh.1219
[55]
Anderson ME, Lefebvre SL, Weese JS (2008) Evaluation of prevalence and risk factors for methicillin-resistant Staphylococcus aureus colonization in veterinary personnel attending an international equine veterinary conference. Vet Microbiol 129: 410–417. doi: 10.1016/j.vetmic.2007.11.031
[56]
Panchaud Y, Gerber V, Rossano A, Perreten V (2010) Bacterial infections in horses: a retrospective study at the University Equine Clinic of Bern. Schweiz Arch Tierheilkd 152: 176–182. doi: 10.1024/0036-7281/a000040
[57]
Weese JS, Lefebvre SL (2007) Risk factors for methicillin-resistant Staphylococcus aureus colonization in horses admitted to a veterinary teaching hospital. Can Vet J 48: 921–926.
[58]
Weese JS, Archambault M, Willey BM, Hearn P, Kreiswirth BN, et al. (2005) Methicillin-resistant Staphylococcus aureus in horses and horse personnel, 2000-2002. Emerg Infect Dis 11: 430–435. doi: 10.3201/eid1103.040481
[59]
Weese JS, Caldwell F, Willey BM, Kreiswirth BN, McGeer A, et al. (2006) An outbreak of methicillin-resistant Staphylococcus aureus skin infections resulting from horse to human transmission in a veterinary hospital. Vet Microbiol 114: 160–164. doi: 10.1016/j.vetmic.2005.11.054
[60]
Porphyre T, Giotis ES, Lloyd DH, Stark KD (2012) A Metapopulation Model to Assess the Capacity of Spread of Meticillin-Resistant Staphylococcus aureus ST398 in Humans. PLoS One 7: e47504. doi: 10.1371/journal.pone.0047504
[61]
Li M, Du X, Villaruz AE, Diep BA, Wang D, et al. (2012) MRSA epidemic linked to a quickly spreading colonization and virulence determinant. Nat Med 18: 816–819. doi: 10.1038/nm.2692
[62]
McAdam PR, Templeton KE, Edwards GF, Holden MT, Feil EJ, et al. (2012) Molecular tracing of the emergence, adaptation, and transmission of hospital-associated methicillin-resistant Staphylococcus aureus. Proc Natl Acad Sci U S A 109: 9107–9112. doi: 10.1073/pnas.1202869109
[63]
Strommenger B, Bartels MD, Kurt K, Layer F, Rohde SM, et al. (2013) Evolution of methicillin-resistant Staphylococcus aureus towards increasing resistance. J Antimicrob Chemother.
[64]
Nübel U, Nitsche A, Layer F, Strommenger B, Witte W (2012) Single-nucleotide polymorphism genotyping identifies a locally endemic clone of methicillin-resistant Staphylococcus aureus. PLoS One 7: e32698. doi: 10.1371/journal.pone.0032698
[65]
Stegger M, Liu CM, Larsen J, Soldanova K, Aziz M, et al. (2013) Rapid Differentiation between Livestock-Associated and Livestock-Independent Staphylococcus aureus CC398 Clades. PLoS One 8: e79645. doi: 10.1371/journal.pone.0079645
[66]
Smith TC, Gebreyes WA, Abley MJ, Harper AL, Forshey BM, et al. (2013) Methicillin-resistant Staphylococcus aureus in pigs and farm workers on conventional and antibiotic-free swine farms in the USA. PLoS One 8: e63704. doi: 10.1371/journal.pone.0063704
[67]
Li S, Skov RL, Han X, Larsen AR, Larsen J, et al. (2011) Novel types of staphylococcal cassette chromosome mec elements identified in clonal complex 398 methicillin-resistant Staphylococcus aureus strains. Antimicrob Agents Chemother 55: 3046–3050. doi: 10.1128/aac.01475-10
[68]
McCarthy AJ, van Wamel W, Vandendriessche S, Larsen J, Denis O, et al. (2012) Staphylococcus aureus CC398 clade associated with human-to-human transmission. Appl Environ Microbiol 78: 8845–8848. doi: 10.1128/aem.02398-12
[69]
Hallin M, De Mendonca R, Denis O, Lefort A, El Garch F, et al. (2011) Diversity of accessory genome of human and livestock-associated ST398 methicillin resistant Staphylococcus aureus strains. Infect Genet Evol 11: 290–299. doi: 10.1016/j.meegid.2010.10.021
[70]
Strommenger B, Braulke C, Heuck D, Schmidt C, Pasemann B, et al. (2008) spa Typing of Staphylococcus aureus as a frontline tool in epidemiological typing. J Clin Microbiol 46: 574–581. doi: 10.1128/jcm.01599-07
[71]
Feil EJ (2004) Small change: keeping pace with microevolution. Nat Rev Microbiol 2: 483–495. doi: 10.1038/nrmicro904
[72]
Kurt K, Rasigade JP, Laurent F, Goering RV, Zemlickova H, et al. (2013) Subpopulations of Staphylococcus aureus clonal complex 121 are associated with distinct clinical entities. PLoS One 8: e58155. doi: 10.1371/journal.pone.0058155
[73]
Roumagnac P, Weill FX, Dolecek C, Baker S, Brisse S, et al. (2006) Evolutionary history of Salmonella Typhi. Science 314: 1301–1304. doi: 10.1126/science.1134933
[74]
van Wamel WJ, Rooijakkers SH, Ruyken M, van Kessel KP, van Strijp JA (2006) The innate immune modulators staphylococcal complement inhibitor and chemotaxis inhibitory protein of Staphylococcus aureus are located on beta-hemolysin-converting bacteriophages. J Bacteriol 188: 1310–1315. doi: 10.1128/jb.188.4.1310-1315.2006
[75]
Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, et al. (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 59: 307–321.
[76]
Rozas J (2009) DNA sequence polymorphism analysis using DnaSP. Methods Mol Biol 537: 337–350. doi: 10.1007/978-1-59745-251-9_17
[77]
Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25: 1451–1452. doi: 10.1093/bioinformatics/btp187
[78]
Rozas J, Sanchez-DelBarrio JC, Messeguer X, Rozas R (2003) DnaSP, DNA polymorphism analyses by the coalescent and other methods. Bioinformatics 19: 2496–2497. doi: 10.1093/bioinformatics/btg359
[79]
Drummond AJ, Rambaut A (2007) BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evol Biol 7: 214. doi: 10.1186/1471-2148-7-214