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Background: Coagulase negative staphylococci (CoNS) are important reservoirs of antibiotic resistance genes and associated mobile genetic elements and are believed to contribute to the emergence of successful methicillin resistant Staphylococcus aureus (MRSA) clones. Although, these bacteria have been linked to various ecological niches, little is known about the dissemination and genetic diversity of antibiotic resistant CoNS in general public settings. Methods: Four hundred seventy-nine samples were collected from different non-healthcare/general public settings in various locations (n = 355) and from the hands of volunteers (n = 124) in London UK between April 2013 and Nov Results: Six hundred forty-three staphylococcal isolates belonging to 19 staphylococcal species were identified. Five hundred seventy-two (94%) isolates were resistant to at least one antibiotic, and only 34 isolates were fully susceptible. Sixty-eight (11%) mecA positive staphylococcal isolates were determined in this study. SCCmec types were fully determined for forty-six isolates. Thirteen staphylococci (19%) carried SCCmec V, followed by 8 isolates carrying SCCmec type I (2%), 5 SCCmec type IV (7%), 4 SCCmec type II (6%), 1 SCCmec type III (2%), 1 SCCmec type VI (2%), and 1 SCCmec type VIII (2%). In addition, three isolates harboured a new SCCmec type 1A, which carried combination of class A mec complex and ccr type 1. MLST typing revealed that all S. epidermidis strains possess new MLST types and were assigned the following new sequence types: ST599, ST600, ST600, ST600, ST601, ST602, ST602, ST603, ST604, ST605, ST606, ST607 and ST608. Conclusions: The prevalence of antibiotic resistant staphylococci in general public settings demonstrates that antibiotics in the natural environments contribute to the selection of antibiotic resistant microorganisms. The finding of various SCCmec types in non-healthcare associated environments indicates the complexity of SCCmec. We also report on new MLST types that were assigned for all S. epidermidis isolates, which demonstrates the genetic variability of these isolates. Keywords: CoNS, Antibiotic resistance, SCCmec,MLST * Correspondence: email@example.com School of Biological and Chemical Sciences, Queen Mary University of London, London, UK School of Health, Sport and Biosciences, University of East London, E1 4NS, London, UK Full list of author information is available at the end of the article © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 2 of 10 Background selected based on staphylococci morphology . The col- Staphylococci are the most frequently isolated nosocomial onies were then purified on Nutrient Agar (Oxoid, Basing- pathogens, accounting for 30% of hospital associated infec- stoke, UK). tions . Despite, that the high virulence of S. aureus has been evidenced in many studies , it is believed that co- Identification agulase-negative staphylococci (CoNS) act as an important All isolates were initially screened using Gram staining, reservoir of antimicrobial resistance genes and resist- catalase and coagulase tests. Those that demonstrated po- ance-associated mobile genetic elements, which can tential staphylococci characteristics were identified by transfer between staphylococcal species. Among Matrix-assisted laser desorption ionization time flight other CoNS, S. epidermidis, S. hominis and S. hae- mass-spectroscopy (MALDI-TOF-MS, Microflex LT, Bru- molyticus are often reported to be resistant to mul- ker Daltonics, Coventry, UK) in a positive linear mode tiple antibiotics [3, 4]. (2000–20,000 m/z range) as described previously . The mecA gene responsible for methicillin resist- The resulting spectra were compared with reference spec- ance was first determined in S. aureus, however, many tra by using the Biotyper 3.0 software (Bruker Daltonics, other staphylococcal species were found to also harbour it Coventry, UK). Escherichia. coli DH5α (Bruker Daltonics, . The mecA gene encodes an additional penicillin-bind- Coventry, UK) was used as a standard for calibration and ing protein 2a (PBP 2a), which mediates cell wall synthesis quality control. in the presence of β-lactam antibiotics . Together with its regulators mecI-mecR1 and site specific recombination Antimicrobial susceptibility test genes ccrA and ccrB,the mecA gene, is located on a mobile A panel of 11 antibiotics was used to determine the anti- genetic element known as staphylococcal cassette chromo- biotic susceptibility of all the isolates. The standard disk dif- some mec (SCCmec). A number of studies have demon- fusion method was used to test AM: amoxicillin (10 μg); strated the transfer of mecA gene from coagulase-negative CEP: cefepime (30 μg);CHL:chloramphenicol (30 μg); staphylococcal species to S. aureus in vivo, and thus con- ERY: erythromycin (5 μg); FC: fusidic acid (10 μg); GEN: tributing to more successful S. aureus clones . To date gentamicin (10 μg); MUP: mupirocin (20 μg); OX: oxacillin 11 SCCmec types have been reported based on combina- (1 μg); PEN: penicillin (1 unit); STR: streptomycin (10 μg); tions of mec (A, B, C1, C2 and D) and ccr (AB1, AB2, TET: tetracycline (10 μg);. The susceptible, intermediate re- AB3, AB4 and ccrC) complexes and so called J regions (1, sistant or resistant were determined by the Guidelines for 2, 3) . Susceptibility Testing . The Minimum Inhibitory Con- Traditionally recognised as hospital associated pathogens, centrations (MIC) for oxacillin were additionally evaluated methicillin resistant coagulase negative staphylococci (MR- using “M.I.C. evaluators” (Oxoid Ltd., Basingstoke, UK). CoNS) have recently been linked with a range of ecological niches (community, wildlife and environmental sources) Detection of mecA gene and staphylococcal cassette [10–12]. As a result, today increasing attention is being paid chromosome mec (SCCmec) typing to the rapid spread of MR-CoNS and their role in transmis- The mecA gene was determined by using PCR method sion within the community and non - hospital settings . as described previously . For mecA positive isolates, In this study we demonstrate the dissemination of anti- SCCmec types were determined by evaluating mec and biotic resistance in CoNS isolated from various environ- ccr complexes . mental sites in London, UK. The characterization of mecA gene and the SCCmec elements provide insights into the di- MLST typing of Staphylococcus epidermidis versity of environmental CoNS clones. Multi-locus sequence typing (MLST) was used to deter- mine the sequence types of S. epidermidis . Sequence Methods types were assigned using the S. epidermidis database Isolation (www.mlst.net). Four hundred seventy-nine samples were collected from different environmental sites in various locations (n = 355) and from the hands of volunteers (n =124)inLondonUK Results between April 2013 and Nov 2014. Environmental sites in- Purification of isolates cluded hotels (n = 100), baby care facilities (n =65), hand- A total of 643 staphylococci isolates were recovered bags (n =43), supermarkets (n = 37), restaurants (n =36), in this study, including those from hotels (n = 74), public transport (n =54), and a public library (n =20). All baby care facilities (n = 46), handbags (n =17), super- specimens were plated on Mannitol Salt Agar (Oxoid, markets (n = 89), restaurants (n = 96), public trans- Basingstoke, UK), and then incubated aerobically at 37 °C portation (n = 94), human hands (n = 192) and public for 24–72 h). One or two colonies for each site were libraries (n = 35) (Additional file 1:Table S1). Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 3 of 10 Species determination (80%) among all 19 staphylococcal species, followed by Six hundred forty-three staphylococcal isolates be- S. cohnii (33%), S. haemolyticus (22%), and S. saprothyti- longing to 19 staphylococcal species were identified cus (20%). Other isolates demonstrated relatively lower in this study. This included: S. epidermidis (n = 193), S. carriage of mecA gene, including S.hominis (3%), S.capitis hominis (n = 161), S. capitis (n = 77), S. warneri (n = 63), S. (8%), S. epidermidis (11%), S.warneri (11%), S.pasteuri haemolyticus (n =45), S. pasteuri (n = 33), S. saprophyticus (13%). No mecA gene was found in the remaining 10 spe- (n =20), S. aureus (n = 12), S. simiae (n =10), S. cohnii (n = cies, including S. aureus, S. simiae, S. equorum, S. caprae, 9), S. sciuri (n =5), S. pettenkoferi (n =3), S. auricularis (n = S. xylosus, S. auricularis, S. simulans, S. arlettae S.petten- 2), S. caprae (n =2), S. equorum (n =2), S. lugdunensis (n = koferi,and S. lugdunensis. 2), S. xylosus (n =2), S. arlettae (n =1), and S. simulans (n SCCmec types were fully determined in forty-six isolates. =1). S. epidermidis was the predominant species, followed Twenty-two out of 68 isolates lacked either the mec gene by S. hominis, S. capitis, S. warneri, S. haemolyticus, S. pas- complex or the ccr gene complex. Thirteen staphylococci teuri,and S. saprophyticus However, the occurrence of (19%) carried SCCmec type V, followed by 8 isolates carry- the species varied for different sites. S. epidermidis ing SCCmec type I (2%), 5 isolates SCCmec type IV (7%), 4 was predominant among the isolates recovered from res- isolates SCCmec type II (6%), 1 isolate SCCmec type III taurants, public transport, hands and handbags, whereas S. (2%), 1 isolate SCCmec type VI (2%), and 1 isolate SCCmec hominis was predominant among the isolates recovered type VIII (2%). In addition, three isolates harboured a new from supermarkets, baby care facilities and hotels and S. SCCmec type 1A, which carried combination of class A haemolyticus was predominantly isolated from the library mec complex and ccr type 1. Of the ten isolates that were (Table 1). non-typeable, three carried a combination of class A mec complex and ccrC, six carried a combination of class B Antibiotic susceptibility test results mec and ccrC, and one carried class B mec and ccr type 3 The disc diffusion method was used to test 606 isolates (Table 3). against a panel of 11 antibiotics. 572 (94%) isolates were resistant to at least one antibiotic, and only 34 isolates Multi-locus sequence typing of S. epidermidis were fully susceptible. Resistance to penicillin, and fusi- MLST was performed to determine the housekeeping dic acid was observed in more than 65% of all staphylo- genes of 13 oxacillin resistant and mecA positive S. coccal isolates tested. 202 (33%) isolates were resistant epidermidis. MLST typing revealed that all S. epidermidis to streptomycin, 190 (31%) to erythromycin, 161 (27%) strains possess new MLST types. MLST types of S. epider- to amoxicillin, 98 (16%) to tetracycline, 87 (14%) to midis isolates with in house numbers of 279, 133, 134, mupirocin, 59 (10%) to gentamicin, 48 (8%) cefepime, 36 135, 126, 259, 124, 127, 234, 187, 308, 153 and 191 were (6%) oxacillin, and 21(3%) chloramphenicol (Table 2). respectively assigned as ST599, ST600, ST600, ST600, ST601, ST602, ST602, ST603, ST604, ST605, ST606, mecA gene determination and SCCmec typing results ST607 and ST608 (Table 4). Three S. epidermidis isolates Sixty-eight (11%) mecA positive staphylococcal isolates shared the same sequence types (ST), including S. epider- were determined, however, no MRSA was determined in midis 133, 134 and 135 that were isolated from different this study. S. sciuri had the highest mecA gene carriage sites of a library (DSL) possessed ST600 whereas S. epider- midis 259, and S. epidermidis 124 that had ST602 se- quence type were isolated from the human hands (HH) Table 1 Predominant and common staphylococcal species and different sites of hotels (DSH) respectively. recovered from the human hands and different environmental sites Discussion Sites Predominant species (%) Commonly isolated species (%) Environmental staphylococcal species BCF S. hominis (17%) S. warneri (17%) Although antibiotic resistance is commonly linked to the DSH S. hominis (30%) S. haemolyticus (18%) clinic, recent studies from different ecological niches re- DSL S. haemolyticus (29%) S. epidermidis (26%) vealed multidrug resistant bacteria is widespread in the DSR S. epidermidis (38%) S. hominis (35%) environment [11, 12, 17]. DSS S. hominis (44%) S. epidermidis (29%) We have previously reported on high levels of antibiotic resistance in staphylococci isolated from different environ- DST S. epidermidis (35%) S. capitis (15%) mental/public settings [11, 12]. In this study we evaluated HB S. epidermidis (40%) S. capitis (27%) the dissemination of antibiotic resistant staphylococci re- HH S. epidermidis (36%) S. hominis (23%) covered from a wide range of environmental settings, and BCF baby care facilities, DSH different sites of hotels, DSL different sites of a characterised the carriage of the mecA gene and the diver- library, DSR different sites of restaurants, DSS different sites of supermarkets; DST different sites of transportation facilities, HB handbags, HH human hand sity of SCCmec elements in these isolates. Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 4 of 10 Table 2 Antibiotic susceptibility profile of staphylococci isolates recovered from general public settings Isolates No of isolates Resistance to a panel of 11 antibiotics (%) OX PG MUP CEF GM FC S A E T C S. epidermidis 176 8 7216 9 7 642226 4316 2 S. hominis 152 2 68 9 5 7 66 24 17 38 21 3 S. capitis 73 4 5815 4 1 604723 1410 5 S. haemolyticus 40 10 50 13 23 15 68 73 45 0 25 10 S. warneri 63 3 5417 10 22 595140 2716 2 S. pasteuri 31 6 6913 6 9 692531 4417 3 S. saprophyticus 20 15 90 25 5 10 100 10 25 35 15 10 S. aureus 12 0 83 17 0 58 83 33 50 25 0 8 S. simiae 10 0 10 0 0 0 40 0 0 0 0 0 S. cohnii 9 24 67 11 33 0 78 78 22 56 11 0 S. sciuri 5 60 60 80 0 20 80 80 40 0 0 0 S. pettenkoferi 3 0 33 0 0 0 676733 0 00 S. lugdunensis 2 0 50 0 0 050 0 00 0 0 S. equorum 2 0 50 50 0 0 50 50 50 0 50 0 S. caprae 2 0 100 0 100 100 100 50 50 0 0 0 S. xylosus 2 0 100 50 50 50 100 100 0 0 50 0 S. auricularis 2 0 50 0 50 0 50 0 50 0 0 0 S. arlettae 1 0 100 0 0 0 100 100 100 100 0 0 S. simulans 1 0 100 0 0 0 100 0 0 0 0 0 OX oxacillin (1 μg), PG penicillin G (1 unit), MUP mupirocin (20 μg), CEF cefepime (30 μg), GM gentamicin (10 μg), FC fusidic acid (10 μg), S streptomycin (10 μg), A amoxicillin (10 μg), E erythromycin (5 μg), T tetracycline (10 μg), C chloramphenicol (30 μg) Six hundred and forty-three staphylococci isolates be- higher than those reported in clinical staphylococci isolates longing to 19 species, including S. epidermidis, S. hominis, [21, 23, 24]. It is widely accepted that higher levels of anti- S. haemolyticus, S. capitis, S. warneri, S. pasteuri, S. sapro- biotic resistance in clinical isolates are due to consistent phyticus, S. cohnii, S. aureus, S. simiae, S. sciuri, S. petten- antibiotic exposure . The environment may also con- koferi, S. lugdunensis, S. equorum, S. caprae, S. xylosus, S. tribute to the development of antibiotic resistance in mi- auricularis, S. simulans, and S. arlettae,were identified in croorganisms due to human/ animal therapeutics, sewage, this study. Interestingly, many of the staphylococci species agriculture and industrial use of antibiotics . There- recovered in our study have previously been associated fore, the wide dissemination of multidrug resistant CoNS with the community, preserved food, and wildlife [4, 10]. in non-healthcare associated environments is a disturbing finding. In our study, 94% of staphylococcal isolates were Antibiotic resistance phenotypically resistant to at least 1 antibiotic, 18% were Antibiotic resistance of staphylococci associated with resistant to five or more antibiotics and only 6% staphylo- healthcare settings is well documented, however, little coccal isolates were fully susceptible. The study also re- is known about the antibiotic resistance in staphylococci vealed that the number of isolates resistant to multiple isolated from different ecological niches . In this study, antibiotics varied between the different isolation sites. The the majority of staphylococci were resistant to penicillin least number of multiple antibiotic resistant CoNS isolates (65%) and fusidic acid (66%) (Fig. 1). Despite that 80% of were recovered from the public transport (58%), the high- hospital associated CoNS (across Europe) were reported est was isolated from hotels (78%). to be resistant to oxacillin , only 6% of CoNS were re- sistant to oxacillin in this study. In addition, the levels Methicillin-resistant staphylococci of resistance to chloramphenicol (3%), cefepime (8%), Methicillin resistant staphylococci pose a major public gentamicin (10%), mupirocin (14%), tetracycline (16%), and health threat, and cause severe economic and health con- erythromycin (31%) were lower compared to those reported sequences . Methicillin resistance is determined by the in clinical settings [19–22]. In contrast, the rates of resist- mecA gene, which encodes for penicillin binding protein ance to fusidic acid (66%), amoxicillin (27%) and strepto- 2a (PBP2a) that has a low affinity to β-lactam antibiotics mycin (33%) in environmental staphylococcal isolates were . Hussain et al. assessed the correlation between mecA Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 5 of 10 Table 3 Molecular characterisation and antibiotic resistance of mecA gene positive staphylococci −1 ID Sites Species PG MUP CEF GM FC S A E T C mecA mec ccr SCCmec MIC/OX (mg l ) 71 HH S. capitis SS S S S R S S S S + –– I 0.5 100 DSH S. cohnii R R S S S S R R S S + Class A 5 5A 1 97 BCF S. cohnii R S R S R R S R S S + Class B 1 I 0.25 279 HH S. epidermidis R S S S R S S R S S + Class B 2 IV 2 127 DSH S. epidermidis R S I S S S R R R S + Class C 5 V 2 139 DSR S. epidermidis R R R S R S R R R S + Class C 5 V 2 191 DSS S. epidermidis R S S S R S R S S S + Class B 4 VI 2 153 DSH S. epidermidis R S S S R S S S S S + Class C 5 V 1 187 DSS S. epidermidis R S S S S S S R S S + Class C 5 V 1 134 DSL S. epidermidis* R S S S R R R R R S + Class B 1 I 1 259 HH S. epidermidis R S R S R R R R S S + Class C 5 V 1 135 DSL S. epidermidis* R S S S R R R R R S + Class B 2 IV 0.5 124 DSH S. epidermidis R S R S R R R S R S + Class B 2 IV 0.5 133 DSL S. epidermidis* R S S S R R R R R S + Class B 3 3B 0.5 126 HH S. epidermidis RS I S S R R R S S + –– III 0.5 119 DSH S. epidermidis R S S S S S R I R S + Class C 5 V 0.12 111 BCF S. epidermidis R S S S S S S S S S + Class A 2 II 0.12 202 DST S. epidermidis S R S S R I S S S S + Class B 5 5B 0.12 264 HH S. epidermidis S R S S R R S S S S + Class B 2 IV 0.06 129 DSL S. epidermidis R S S S R R R R R S + Class B 1 I 0.03 362 DSL S. haemolyticus R S I S S R R S S S + Class C 5 V 2 367 DSL S. haemolyticus R S R S S R R S S S + Class C 5 V 2 355 DSH S. haemolyticus R S R R R R R S R S + Class C 5 V 2 384 HH S. haemolyticus R R S S R R S S S S + Class C 5 V 2 322 DSH S. haemolyticus R S S S S I R S R S + Class A 1 1A 0.25 382 HH S. haemolyticus RS I S R R S R S S + –– II 0.25 323 DSH S. haemolyticus S S S R S R R I R S + Class A 2 II 0.12 381 HH S. haemolyticus S S I S S R S S S S + Class B 5 5B 0.12 360 DSH S. haemolyticus S S S S R S S S S R + Class B 5 5B 0.06 369 DSL S. haemolyticus R S S S R R S R S R + Class B 1 I 0.03 413 DSH S. hominis S R S S R R S S S S + Class C 5 V 2 506 DSS S. hominis S S S S R S S S S S + Class B 1 I 0.5 400 DSH S. hominis R R S S R S R R R S + Class A 1 1A 0.12 326 DSH S. hominis S S S S S S R I S S + Class A 1 1A 0.06 589 HB S. pasteuri R S S S R R S R S S + Class A 5 5A 0.25 592 HH S. pasteuri S R S S R R S S S S + Class B 5 5B 0.25 627 HH S. saprophyticus R I S S R S S S R S + Class B 5 5B 0.5 621 DSS S. saprophyticus R R R S R S R S R S + Class B 2 IV 0.25 630 HH S. sciuri R R I S R R S S S S + Class A 4 VIII 2 632 DSH S. sciuri R S I S R R R S S S + Class A 5 5A 1 633 DSH S. sciuri R R I R R R R S S S + Class B 5 5B 1 629 HH S. sciuri SR S S S S S S S S + –– II 0.25 704 HH S. warneri R S I S R R R R S S + Class C 5 V 0.5 662 DSH S. warneri R S S R R R R S R S + Class C 5 V 0.25 Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 6 of 10 Table 3 Molecular characterisation and antibiotic resistance of mecA gene positive staphylococci (Continued) −1 ID Sites Species PG MUP CEF GM FC S A E T C mecA mec ccr SCCmec MIC/OX (mg l ) 694 HH S. warneri RS S S S S S S R S + –– I 0.25 655 BCF S. warneri S R S R R R S I S S + Class B 1 I 0.12 Note: * S. epidermidis isolates with similar MLST types R: resistant, S sensitive. I intermediate BCF baby care facility, DSH different sites of hotels, DSL different sites of a library, DSR different sites of restaurants, DSS different sites of supermarkets, DST different sites of transportation facilities, HB handbags, HH human hands A amoxicillin (10 μg), CEF cefepime (30 μg), C chloramphenicol (30 μg), E erythromycin (5 μg), FC fusidic acid (10 μg), GM gentamicin (10 μg), MUP mupirocin (20 μg), OX oxacillin (1 μg), PG penicillin G (1 unit), S streptomycin (10 μg), T tetracycline (10 μg) − 1 gene and oxacillin susceptibility breakpoints (0.5 mg l ) induced high levels of oxacillin resistance was determined of 493 clinical CoNS belonging to and classified into 4 cat- in OS-MRSA [30, 31]. In this study, 68 (46%) staphylococ- egories . The mecA gene positive staphylococci were cal isolates were confirmed by PCR to carry the mecA gene, categorized into groups I and II, and demonstrated that however, they were phenotypically susceptible to oxacillin − 1 group I (S. haemolyticus (83.3%), S. epidermidis (61.9%), S. with the MICs (oxacillin) varying from 0.015 to 2 mg l . hominis (51.8%)) differs from group II (S. cohnii (28.5%), S. This study demonstrates the prevalence of mecA positive warneri (27.3%), S. saprophyticus (9.0%)) by their high levels but oxacillin susceptible CoNS (OS-CoNS) in the environ- of mecA-carriage . Interestingly, S. hominis (38%), S. ment. Little is known about OS-CoNS isolates recovered haemolyticus (22%), and S. epidermidis (7%) isolated in this from the environment and their epidemiological data are study harboured significantly lower levels of the mecA gene. limited. Additional studies are necessary to further our un- Moreover, in this study S. cohnii (33%) and S. saprophyticus derstanding of the prevalence and molecular epidemiology (10%) showed higher mecA gene carriage than clinical iso- of OS-CoNS in the environment. lates reported by Hussain, et al. , whereas the levels of mecA gene carriage in S. warneri (6%) were lower than in SCCmec elements clinical isolates. No mecA gene was detected in staphylo- SCCmec is a mobile genetic element with two essential coccal species of groups III and IV, which included S. xylo- components: the mec gene complex, and the cassette sus, S. lugdunensis, S. capitis, S.simulans,and S. schleiferi chromosome recombinase (ccr) gene complex . The . Similarly, in this study S. lugdunensis, S. xylosus and S. combination of the mec gene complex and ccr gene com- simulans were determined to be susceptible to oxacillin plex confers different SCCmec types . SCCmec type I, and lacked mecA gene. However, in contrast to the reports II, III are reported to be associated with MRSA recovered by Hussain, et al. wefound that mecA gene was from healthcare settings, whereas SCCmec type IV and V present in 8% of S. capitis isolates. are mainly associated with the community . Moreover, Oxacillin susceptible mecA gene positive S. aureus (OS- it has been shown that the size of SCCmec types IV and V MRSA) has been reported worldwide, and the risk of are smaller than SCCmec types I, II and III, thus conferring Table 4 MLST types of 13 oxacillin resistant and mecA positive S. epidermidis ID Sites Species arcC aroE gtr mutS pyrR tpiA yqiL MLST types 279 HH S. epidermidis 57 17 5 5 3 4 31 ST599 133 DSL S. epidermidis 57 1 2 2 4 1 4 ST600 134 DSL S. epidermidis 57 1 2 2 4 1 4 ST600 135 DSL S. epidermidis 57 1 2 2 4 1 4 ST600 126 HH S. epidermidis 57 25 9 5 6 1 8 ST601 259 HH S. epidermidis 57 1 2 2 4 1 1 ST602 124 DSH S. epidermidis 57 1 2 2 4 1 1 ST602 127 DSH S. epidermidis 57 10 5 5 10 16 21 ST603 234 HB S. epidermidis 57 1 1 1 2 41 1 ST604 187 DSS S. epidermidis 57 1 1 2 2 1 1 ST605 308 HH S. epidermidis 57 1 2 2 4 7 1 ST606 153 DSH S. epidermidis 57 1 22 2 2 16 1 ST607 191 DSS S. epidermidis 57 3 5 5 7 14 11 ST608 HH human hands, DSL different sites of a library, DSH different sites of hotels, DSS different sites of supermarkets MLST Multi-locus sequence typing Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 7 of 10 Fig. 1 The scatter plot of staphylococcal (> 30 isolates) susceptibility profile. Legend: OX: oxacillin (1 μg);PG: penicillin G (1unit);MUP: mupirocin (20 μg); CEF: cefepime (30 μg); GM: gentamicin (10 μg); FC: fusidic acid (10 μg); S: streptomycin (10 μg);A: amoxicillin (10 μg); E: erythromycin (5 μg); T: tetracycline (10 μg); C: chloramphenicol (30 μg) increased mobility by their smaller size and contributing lacks ccr,while,SCCmec12263 is reported to carry the ccr the spread of these smaller SCCmec elements .In this complex but lacks mec complex [36, 37]. In this study, study, SCCmec type I, II or III were found in 19% (n =13) 21 isolates (29%) were categorized as (ψ)-SCCmec and of mecA-positive CoNS, whereas 27% (n =18) of CoNS SCCmec12263 since they lacked either mec complex or were determined to harbour SCCmec type IV or V. ccr complexes. ψ SCC element is characterized by lacking SCCmec type VI and VIII were previously identified in genes for ccr and mec . One of S. saprophyticus isolates Portugal (2006) and Canada (2009) in hospital associated in this study was found to possess the ψ SCC element MRSA (HA-MRSA) [33, 34]. In this study, we identified (Table 5). one of each type, however, we did not detect SCCmec types IX. MLST of S. epidermidis Becker et al., have previously summarized the commu- Whilst many studies have reported on the changing epi- nity and livestock associated staphylococcal species and demiology of S. aureus, epidemiological data of other their SCCmec types, which included S. capitis (I, IA, II, III, staphylococcal species are limited [38, 39]. In this study, 10 IV, IVa, V, non-typeable: (NT)), S. cohnii (NT), S. epider- new MLST types were determined in 13 S. epidermidis iso- midis (I, IIa, IIb, III, III (variant), IV, IVa, IVb, IVc, IVd, lates. Interestingly, although isolates recovered from human IVe, IVg, V, VI, NT), S. haemolyticus (I, II, II.1, III, III hands (S. epidermidis 259/ SCCmec V) and hotels (S. epi- (variant), IV, V, NT), S. honomis (I, III, IV, NT), S. pasteuri dermidis 124/ SCCmec IV) harboured different SCCmec (IVc), S. saprophyticus (III, NT), S. sciuri (I, III, IIIA, V, types, they shared the same MLST type ST602. In addition, VII, NT) and S. warneri (IV, IV.1, IVb, IVE) . In this three S. epidermidis isolates recovered from libraries (S. epi- study, species associated SCCmec types differed and in- dermidis 133, S. epidermidis 134, S. epidermidis 135) shared cluded the following: S. capitis (I, NT), S. haemolyticus (I, the same MLST type ST600 (Table 4). However, despite II, V, NT) and S. hominis (I, V, NT), S. cohnii (I, V, NT), S. sharing the same MLST type S. epidermidis 133, S. epider- pasteuri (NT), S. saprophyticus (IV, NT), S. sciuri (II, VIII), midis 134 and S. epidermidis 135 harbored SCCmec type S. warneri (I, V, NT). S. epidermidis possessed SCCmec 3B, I, IV respectively. Others reported that S. epidermidis types similar to those reported previously . ST2 was associated with type II, III, IV and non-typable Thirteen unclassified SCCmec types were determined in SCCmec,and S. epidermidis ST22 harboured SCCmec type this study, including three carrying class A mec complex III, IV and V . and ccrC, six had a combination of class B mec and ccrC, one carried class B mec and ccr3, and three had a combin- Conclusions ation of class A mec complex and ccr type 1. The 1A was Systematic analysis of staphylococci isolated from previously defined as a new SCCmec type 1A by others non-healthcare environments provided insights into . Pseudo (ψ)-SCCmec harbours the mec complex but the diversity and antibiotic susceptibility patterns of these Xu et al. Antimicrobial Resistance and Infection Control (2018) 7:73 Page 8 of 10 Table 5 The diversity of SCCmec types of mecA gene positive staphylococci −1 ID Sites Species PG MUP CEF GM FC S A E T C mecA mec ccr SCCmec MIC/OX (mg l ) 75 HH S. capitis R S S S R R S S S S + Class A NT Pseudo (ψ)-SCCmec 0.5 81 HH S. capitis R S R S R R R R S S + NT 5 SCCmec12263 0.5 70 HH S. capitis R S S S S R S S R S + NT 5 SCCmec12263 0.25 83 HH S. capitis S R S S R R S S S S + NT 5 SCCmec12263 0.12 24 DSH S. capitis S S S S R R S S S S + NT 1 SCCmec12263 0.12 108 HH S. cohnii S S I S R R S R R S + Class A NT Pseudo (ψ)-SCCmec 1 308 HH S. epidermidis R R S S R S R R S S + Class B NT Pseudo (ψ)-SCCmec 2 234 HB S. epidermidis S R S S R R S R R S + Class A NT Pseudo (ψ)-SCCmec 1 249 DSH S. epidermidis R S S S R R S S R S + NT 2 SCCmec12263 0.12 125 DSH S. epidermidis S S I S S R S S S S + NT 5 SCCmec12263 0.06 185 DSS S. epidermidis R S S S S S S S S S + Class C NT Pseudo (ψ)-SCCmec 0.06 498 DSS S. hominis R S S S R S S R S S + Class A NT Pseudo (ψ)-SCCmec 0.5 426 DSH S. hominis R S I S R R R R S S + Class A NT Pseudo (ψ)-SCCmec 0.25 412 DSH S. hominis R S S S R S R R S S + NT 1 SCCmec12263 0.06 391 BCF S. hominis R S S S R S S S S S + NT 5 SCCmec12263 0.03 593 HH S. pasteuri R S S R R R R S S S + NT 5 SCCmec12263 0.5 597 HH S. pasteuri R R I S S R S S S S + NT 5 SCCmec12263 0.5 616 BCF S. saprophyticus R R S S R I R R R S + NT 5 SCCmec12263 256 612 BCF S. saprophyticus RR S S R S S R S S + NT NT ψ SCC 1 659 DSH S. warneri R R S S R R R S S S + NT 5 SCCmec12263 0.5 648 BCF S. warneri R S S R R S R S S S + NT 5 SCCmec12263 0.06 645 BCF S. warneri R S S S R S S S S S + NT 4 SCCmec12263 0.015 R resistant, S sensitive, I intermediate BCF baby care facility, DSH different sites of hotels, DSL different sites of a library, DSR different sites of restaurants, DSS different sites of supermarkets; DST different sites of transportation facilities, HB handbags, HH human hands A amoxicillin (10 μg), CEF cefepime (30 μg), C chloramphenicol (30 μg), E erythromycin (5 μg), FC fusidic acid (10 μg), GM gentamicin (10 μg), MUP mupirocin (20 μg), OX oxacillin (1 μg), PG penicillin G (1 unit), S streptomycin (10 μg), T tetracycline (10 μg) isolates. Multi-drug resistance was commonly seen in Abbreviations AM: Amoxicillin; BCF: Baby care facility; CEP: Cefepime; each staphylococcal species. The prevalence of mul- CHL: Chloramphenicol; CoNS: Coagulase-negative staphylococci; tiple antibiotic resistant staphylococci in this study DSH: Different sites of hotels; DSL: Different sites of a library; DSR: Different provides evidence that antibiotics in the natural en- sites of restaurants; DSS: Different sites of supermarkets; DST: Different sites of transportation facilities; ERY: Erythromycin; FC: Fusidic acid; vironments can contribute to the selection of anti- GEN: Gentamicin; HB: Handbags; HH: Human hands; MALDI-TOF-MS: Matrix- biotic resistance in microorganisms. The finding of assisted laser desorption ionization time flight mass-spectroscopy; various SCCmec types in non-healthcare associated en- MIC: Minimum Inhibitory Concentrations; MLST: Multi-locus sequence typing; MR-CoNS: Methicillin resistant coagulase negative staphylococci; vironments emphasizes the complexity of SCCmec ele- MRSA: Methicillin resistant Staphylococcus aureus; MUP: Mupirocin; ments. In addition to this, we also report on new MLST OX: Oxacillin; PEN: Penicillin; SCCmec: Staphylococcal cassette chromosome types that were assigned for all S. epidermidis isolates. This mec; ST: Sequence types; STR: Streptomycin; TET: Tetracycline highlights the genetic variability of these isolates. In conclu- Funding sion, the non-healthcare environments may act as a reser- This work was part of Zhen Xu’s PhD study funded by China Scholarship voir of multidrug resistant staphylococci, and current Council. infection control measures are ineffective in limiting the Availability of data and materials spread of these bacteria. All data generated or analysed during this study are included in this published article and its supplementary information files. Additional file Authors’ contributions ZX: samples collection, laboratory work, data analysis, manuscript preparation. HS: study design, critically reviewing the paper. RM: Data Additional file 1: Table S1. Isolates collected from different analysis, critically reviewing the paper. JC: data analysis, critically reviewing environmental sites and human hands (PDF 46 kb) the paper. WZ: data analysis, critically reviewing the paper. YL: data analysis, Xu et al. 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Antimicrobial Resistance and Infection Control – Springer Journals
Published: Jun 13, 2018
Keywords: CoNS; Antibiotic resistance; SCCmec; MLST
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