Présence, selon l origine du réservoir humain ou animal, des gènes codant pour l immune evasion cluster genes, dans différentes lignées clonales de Staphylococcus aureus Vandendriessche S, Deplano A, Nonhoff C, Dodemont M, Roisin S, R De Mendonça and Denis O Centre National de Référence Staphylococcus aureus, Belgium
Epidemiological history of S. aureus Occassional infection in companion/food-producing animals transfer human to animal S. aureus = human pathogen Dramatic change MRSA CC398 in pigs!!!! Occassional infection in farmer or family transfer animal to human Prompted research on MRSA/S. aureus from animal origin
S. aureus from animal origin Animal-derived S. aureus strains: spa, MLST, micro-array, SNPs, WGS, 2 trends: Lineages unique to animals: CC151, CC133, CC130, CC873 Lineages shared between H & A: CC1, CC5, CC30, CC97, CC398 CC398, CC5 CC97
Host adaptation Host jump (H A) is followed by host adaptation Common themes: Gene decay: spa in poultry S. aureus (Lowder, PNAS 2009) Allelic diversification: vwbp adapted to ruminants (Guinane, GBE 2010) Loss/acquisition of genes: IEC genes Immune Evasion Cluster (IEC) genes: Carried on MGE (φ3 prophage) scn (SCIN), sak (staphylokinase) and chp (CHIPS) Strict human specificity (Rooijakkers, Nature Immun 2005) Found in 95% of S. aureus from human origin, but nearly absent in S. aureus from animals (Verkaik, CMI 2001; Price, mbio 2012) IEC essential for colonization of humans but not animals
Objectives 1) Assess the presence of S. aureus lineages previously detected in food producing animals among a collection of human-derived S. aureus (from National Reference Centre for S. aureus, NRCS) 1) To compare the prevalence of immune evasion cluster (IEC) genes in this human-derived subset with the prevalence in S. aureus isolates from livestock and farmers.
Strain collections Two animal-derived collections 1) Nasal S. aureus isolates from farmers & livestock (pig, bovine, broiler) MRSA CC398, CC97 MSSA CC1, CC5, CC9, CC30, CC45, CC97, CC398, CC133, 2) S. aureus isolates causing serious infections in Belgian rabbitries MSSA CC121 A human-derived strain collection 3) S. aureus sent to NRCS ( 08-13) for toxin detection/molecular typing Present work: Characterization of S. aureus isolates assigned to a lineage/spa type previously detected among livestock (Vandendriessche et al., JAC 2013 and JAC 2014, Vancraeynest et al., JVM 2006)
Workflow 3 collections 2008-2013: 1639 S. aureus isolated from patients, sent to NRCS for toxin/typing spa-typing BURP-cluster Isolates assigned to lineage/spatype found in livestock (n = 218) S. aureus from livestock farms representative subset (n = 195; 119 MRSA & 76 MSSA) S. aureus from rabbits (n = 7 MSSA) Collection 1 2 3 Further characterization: - Toxin detection (PVL/TSST-1) - Antimicrobial susceptibility testing - Presence of IEC genes by (in-house) PCR
Genetic diversity 13,3% of patient isolates assigned to spa-type/lineage prev. detected among livestock S. aureus from farms & rabbits n = 195 + 7 5% 5% CC1-t127 3% 3% 2% 5% 3% CC9 5% 3% 65% SHARED CC30-t021/t318 Focus CC45-t095 on CC398 CC97 55% CC121-t272/t645 CC398 CC133 CC5-t3478 CC705 other UNIQUE TO ANIMALS S. aureus from patients n = 218 17% 4% 0% 3% CC398 - MRSA (77,5%) - MSSA (22,5%) 1% 20%
Geographic distribution of CC398 Methicillin resistance 14 85% of MRSA CC398 is found in Flanders t011 (85%) 43 14/1 6/15 6 5/3 1 6 MRSA CC398 MSSA CC398 Flanders; intensive A-farming Wallonia; extensive A-farming Brussels/Capital : no farming 1/2 1 96% of MSSA CC398 is from Brussels/Wall t571 (54%) t034 (19%) t1451 (19%)
Geographic distribution of CC398 IEC genes 97% of CC398 in Flanders is IEC - 14 42/1 14/1 9/12 6 4/4 1 6 CC398-IEC - CC398-IEC + 1/2 Flanders; intensive A-farming Wallonia; extensive A-farming Brussels/Capital : no farming 1 96% of MSSA CC398 is IEC +
Comparison of MRSA & MSSA CC398 MRSA CC398 MSSA CC398 Most frequent spa types t011 t571, t034, t1451 Origin Screening 47% 15% (outbreak) Blood 10% 0% SSTI 22% 74% Respiratory tract 16% 4% Other 5% 7% Median age (range) 53yr (0-92) 29yr (0-83) Livestock contact 80% (for n=42) No data Presence of IEC 3% 96% Toxin genes 0% 26% Tetracycline R, tet(m) 100% 0% Erythromycine R 41% 81% Multiresistance AG, cip, Trim Infrequent
Two CC398 subclones International phenomenon Start: MRSA CC398 in livestock, occassional infection in persons in contact Later: infections by MSSA CC398 in patients without livestock contact worldwide: Europe, USA, South-America, Asia different settings: hospital, community, jail, Both subclone differ in - carriage of IEC genes, - Resistance profile (tetracycline), - Interhuman transmissibility, - Toxin gene content (PVL),
Two CC398 subclones International phenomenon Start: MRSA CC398 in livestock, occassional infection in persons in contact Later: infections by MSSA CC398 in patients without livestock contact worldwide: Europe, USA, South-America, Asia different settings: hospital, community, jail, MSSA CC398 is frequent in France link to epidemiology in Belgium?
Lineages shared between H & A CC1-t127 Animal-derived Patient ID 5 MSSA 18 MRSA, 15 MSSA Presence Pig, veal, child on pig farm patient PVL/TSST No PVL: 30% IEC 3/5 - (cluster: pig & child) 2/5 + 97% + al Italy, Spain, Cyprus; DNA-micro-array distinction of porcine & human cluster e.g. different in hlb & sak
Lineages shared between H & A CC9 CC30 Animal-derived Patient ID 11 MSSA 3 MSSA spa types t337, t1430, t1939, t2315 t587 Presence pigs, human on pig farm patients PVL/TSST no no IEC 100% - 2/3 - (mother & child) Animal-derived Patient ID 5 MSSA 10 MRSA, 34 MSSA spa types t021, t318, t1298, t3427 t021, t318 Presence pigs patients PVL/TSST no PVL: 66%; TSST: 25% IEC 100% - 97% +
Lineages shared between H & A CC97 Animal-derived Patient ID 2 MRSA, 8 MSSA 2 MRSA, 4 MSSA spa types t267, t1236, t1247, t1730, t2421, t2802, t5727 t224, t267, t359, t2421, t2802 Presence Veal, Beef, Dairy, human on dairy farm patients PVL/TSST no no IEC 70% - 30% + (cluster: dairy & human) 100% + CC45 1 MSSA t095 in veal calf, 1 MRSA t095 in patient; both IEC +
Lineages shared between H & A Isolates from commercial rabbitries facing chronic problems of staphylococcosis (pododermatitis, abscesses & mastitis) CC121 Animal-derived Patient ID 6 MSSA 8 MSSA spa types t272, t645, t741 t645, t272 Presence rabbits patients PVL/TSST no no, but 100% eta/etb + IEC 100% - 7/8 + 1/8 -
Lineages unique to animals Some lineages were frequent in livestock, but absent in the patient collection spa types t3478 t1403 t8662 CC5 CC133 CC705 Presence Broilers Dairy cattle Beef cattle Human on beef farm t529 Toxins none none none Dairy cattle Beef cattle IEC 100% negative 100% negative 100% negative
Conclusion Epidemiological & molecular evidence for two distinct CC398 subclones in Belgium Livestock-dependent, MRSA, IEC Livestock-independent, MSSA, IEC + Other lineages, such as CC1, CC9, CC30, CC97 & C121 variably carry IEC genes correlation with a human or animal origin Lineages adapted to different reservoirs Monitoring is important!!
Acknowledgements Centre Nationale de Référence S. aureus, Hôpital Erasme, ULB Olivier Denis Ariane Deplano Claire Nonhoff Magali Dodemont Ricardo de Mendonça Sandrine Roisin Maria Argudin Université de Gand, Faculté des véterinaires Freddy Haesebrouck Katleen Hermans THANK YOU FOR YOUR ATTENTION