Methicillin-resistant Staphylococcus Aureus ST398 Research Articles

PFGE diversity within the methicillin-resistant Staphylococcus aureus clonal lineage ST398

BackgroundLivestock has recently been identified as a new reservoir of methicillin-resistant Staphylococcus aureus (MRSA). Most isolates belong to ST398 and are non-typeable with PFGE using SmaI, making it difficult to study transmission and outbreaks. Therefore, a new PFGE using Cfr9I, a neoschizomer of SmaI was optimized and evaluated to investigate ST398 isolates.ResultsAfter optimizing and evaluating the Cfr9I PFGE, clear and reproducible banding patterns were obtained from all previously non-typeable MRSA (NTSmaI -MRSA) isolates. The PFGE patterns of ST398 isolates showed more diversity than with spa-typing and/or MLST. The PFGE results showed diversity within and between the two most prevalent spa-types of NTSmaI -MRSA (t011 and t108). No match was found, when comparing banding patterns of the NTSmaI -MRSA with 700 different PFGE types, obtained with SmaI digestion, in our database of more than 4000 strains. Furthermore, possible transmission among veterinarians and their family members was investigated and an outbreak of ST398 MRSA in a residential care facility was confirmed with the Cfr9I PFGE.ConclusionsThe adjusted PFGE can be used as a method for selecting important and distinct ST398 isolates for further research. The adjustments in the PFGE protocol using Cfr9I are easy to implement to study the ST398 clonal lineage in laboratories which already have a PFGE facility.

Emergence of methicillin-resistant Staphylococcus aureus (MRSA) in different animal species

The emergence of methicillin-resistant Staphylococcus aureus (MRSA) in animals such as horses, pet animals and productive livestock has raised questions of a probable human origin and in more general of host specificity of S. aureus. Particular clonal lineages are obviously specific for humans (e.g. ST15, ST25, ST45) and other for ruminants (e.g. ST151). MRSA associated with veterinary nosocomial infections (e.g. ST8 and ST254 in horses, ST22 in small animals) very likely have their origin in health care facilities. MRSA ST398 which became first known from widespread colonization in industrially raised pigs seems to have a limited host specificity and is able to colonize and to cause infections in various hosts. Mechanisms of host adaptation and their genomic background are poorly understood so far.

Heterogeneity among methicillin-resistant Staphylococcus aureus from Italian pig finishing holdings

A survey for methicillin-resistant Staphylococcus aureus (MRSA) in finishing pig holdings was carried out in Italy in 2008. MRSA isolates were characterised by spa-, MLST-, SCCmec- and antimicrobial susceptibility typing. A prevalence of 38% (45/118, 95% CI 29.4–46.9%) positive holdings was observed. Eleven different spa-types were found among 102 MRSA isolates, clustering in lineages associated with farm animals (ST398, ST9, ST(CC)97 in 36 holdings) and humans (ST1, 7 holdings). Nine (7.6%) holdings were positive for two, three or four different and unrelated spa-types in various combinations. ST398 was the most prevalent lineage (33 positive holdings). The most prevalent spa-type was t899 (ST398), detected in 22 positive holdings. Three novel spa-types (t4794 of ST9; t4795 of ST97; t4838 of ST398) were detected. Ten holdings were positive for spa-type t1730, that proved to be a new single-locus variant of ST97, within the CC97 (ST1476). The most prevalent SCCmec was Type V (79 isolates), while Type IVb was found in 10 isolates. None of the isolates was positive for Panton-Valentine Leukocidin, while most of the t127 and t1730 isolates, one t4794, one t4795, and one t2922 were positive for LukE-LukD genes. All 64 antimicrobial susceptibility tested isolates were resistant to tetracyclines, with high resistance rates to trimethoprim (68.8%), erythromycin (60.9%), and ciprofloxacin (35.4%). All t127, ST1 isolates were resistant to tetracycline–ciprofloxacin–erythromycin. This survey provides the first report of MRSA ST1 and ST(CC)97 among pigs and the first report of MRSA ST9 from pigs in Europe. The presence of human-associated CA-MRSA (t127, ST1, SCCmec type V) in 6% holdings surveyed can represent an additional MRSA reservoir for infections in humans.

Occurrence and genotyping using automated repetitive-sequence–based PCR of methicillin-resistant Staphylococcus aureus ST398 in Southeast Austria

In this retrospective study, the occurrence and genetic relatedness of methicillin-resistant Staphylococcus aureus (MRSA) ST398 in Austrian MRSA patients was investigated. From 2002 to 2008, 14 MRSA ST398 were detected. First occurrence of MRSA ST398 was already found in 2004. Spa ribotyping assigned 12 isolates to spa type t011 and 1 each to spa type t034 and spa type t1451. Isolated MRSA ST398 was nontypeable by pulsed-field gel electrophoresis (NT-MRSA) using restriction enzyme SmaI; therefore, genotyping was performed using automated repetitive-sequence–based PCR (rep-PCR) on the DiversiLab system. Rep-PCR results assigned 10 (71%) of the 14 MRSA ST398 into 1 cluster with a similarity >95%; there was 1 cluster consisting of 2 isolates with a similarity >99% and 2 unique MRSA ST398 isolates. In conclusion, MRSA ST398 was continuously detected in Southeast Austria; first in 2004 with up to 5 MRSA ST398 isolates in 2008. Automated rep-PCR proved as a reliable technique in determining genetic relatedness of NT-MRSA ST398 and demonstrates clonal spread of MRSA ST398 in the investigated region.

Methicillin-ResistantStaphylococcus aureusST398 in Swine Farm Personnel, Belgium

We assessed methicillin-resistant Staphylococcus aureus (MRSA) in persons on 49 swine farms in Belgium. Surveys showed that 48 (37.8%) persons carried MRSA ST398 and 1 (0.8%) had concurrent skin infection. Risk factors for carriage were MRSA carriage by pigs, regular contact with pigs and companion animals, and use of protective clothing.

Unexpected sequence types in livestock associated methicillin-resistant Staphylococcus aureus (MRSA): MRSA ST9 and a single locus variant of ST9 in pig farming in China

In October 2008 nine farrow-to-finish pig farms were visited in Shuangliu County in Sichuan Province, China. One farm was empty for one month but not cleaned after depopulation. Dust samples were collected at each farm and analysed for the presence of methicillin-resistant Staphylococcus aureus (MRSA). Dust samples from four farms were also analysed for the presence of methicillin-susceptible S. aureus (MSSA). On 5/9 farms MRSA was isolated and on 2/4 farms MSSA was isolated. On two farms, including the empty farm, no MRSA or MSSA could be detected. All MRSA isolates (n=43) belonged to spa type t899. MSSA isolates belonged to spa type t899 (n=12) and spa type t034 (n=2). From 4/9 farms the MRSA isolates of spa type t899 were assigned to multilocus sequence type (MLST) ST9 whereas on one farm the MRSA spa type t899 isolates belonged to a single locus variant of MLST ST9 (ST1376). MSSA isolates with spa type t899 belonged to MLST ST9 and the MSSA with spa type t034 belonged to MLST ST398.This is the first report on MRSA in pig farms in China and the first time that MRSA ST9 and a single locus variant of ST9 are detected in pig farms. This study shows that livestock associated MRSA is not restricted to clonal lineage ST398 as found in Europe and Northern America in commercial pigs but that other MRSA lineages are able to spread in livestock as well. The study confirms that livestock may act as a reservoir for MRSA.

Methicillin-ResistantStaphylococcus aureusST398 from Human Patients, Upper Austria

Methicillin-resistant Staphylococcus aureus (MRSA) clonal type ST398 is usually associated with animals. We examined 1,098 confirmed MRSA samples from human patients and found that 21 were MRSA ST398. Most (16) patients were farmers. Increasing prevalence from 1.3% (2006) to 2.5% (2008) shows emergence of MRSA ST398 in humans in Austria.

Epidemiology and susceptibilities of methicillin-resistant Staphylococcus aureus in Taiwan: emphasis on chlorhexidine susceptibility

Chlorhexidine is an antiseptic agent used for hand hygiene worldwide. To evaluate the susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) to chlorhexidine, this study determined MICs of chlorhexidine and another 12 antimicrobial agents, carriage of the Panton–Valentine leukocidin, qacA/B, and smr genes, genetic relatedness by multilocus sequence typing (MLST), and staphylococcal cassette chromosome mec element type for 206 MRSA clinical isolates from the Taiwan Surveillance of Antimicrobial Resistance program III and IV (years 2002 and 2004) from 26 hospitals. Using MLST, we respectively identified 102 (49.5%), 68 (33.0%), 13 (6.3%), 5 (2.4%), 5 (2.4%), and 13 (6.3%) isolates as ST239, ST59, ST5, ST241, ST573, and other types. The MIC 50 and MIC 90 of chlorhexidine for all 206 isolates were 2 and 8 μg/mL, respectively. Seventy-three (35.4%) isolates carried qacA/B gene, but none carried smr. For the 72 (35.0%) MRSA isolates with chlorhexidine MIC ≥4 μg/mL, 53 were ST239 (49 of them carried qacA gene), 12 were ST5 (all carried qacB gene), 5 were ST241 (4 carried qacA gene), 1 was ST338 (and carried qacA gene), and 1 was ST573 (and carried qacA gene). Compared with other sequence-type MRSA isolates, ST239 MRSA isolates were the most resistant to both chlorhexidine and other antimicrobial agents. Methicillin-resistant S. aureus strains with disinfectant resistance qacA/B genes are common in Taiwan. High frequency of qacA/B genes among specific sequence types (ST239, ST5, and ST241) resulted in low susceptibility to chlorhexidine. Periodic surveillance of antiseptic susceptibility among MRSA isolates is important for the control of nosocomial hospital-acquired infections.

SCCmecTyping in Methicillin-ResistantStaphylococcus aureusStrains of Animal Origin

We thank Jansen et al. for their comments about the SCCmec types of sequence type (ST) 398 methicillin-resistant Staphylococcus aureus (MRSA) isolates (1). For SCCmec typing of MRSA, several different PCR methods have been published. We originally chose the SCCmec PCR developed by Zhang et al. (2) because at that time it was the method of choice in many published papers. Fluit et al. questioned whether the SCCmec type III isolates were correctly typed (1). To prove that the results of typing these 4 isolates were incorrect, these researchers performed several different SCCmec PCRs, including a PCR with primers they developed themselves. In addition, Southern hybridization was done. The results showed that SCCmec III ST398 MRSA isolates should be typed as SCCmec type V. In this conclusion we agree with the authors. It seems clear that Zhang’s method incorrectly identified 4 of the animal-related ST398 isolates as SCCmec type III instead of SCCmec type V. Whether all ST398 MRSA are SCCmec type IV or V remains unclear. Recently, an article by Nemati et al. was published in which ST398 MRSA was also typed as SCCmec III (3). However, in that study the SCCmec typing method of Zhang was also used. In conclusion, the choice of SCCmec typing method is directly related to obtaining accurate SCCmec results for ST398 isolates. To date, almost all animal-related ST398 MRSA isolates are SCCmec types IV and V.

Clusters of Infections in Horses with MRSA ST1, ST254, and ST398 in a Veterinary Hospital

During 2006 and 2007 small clusters of methicillin-resistant Staphylococcus aureus (MRSA) infections in horses were recorded in different clinical departments of a veterinary university. The infections were caused by different MRSA clones (ST1, ST254, and ST398). In the same time, nasal colonization of veterinarians, veterinary personnel, and students was observed indicating transmission to humans.

Comparison of Adhesion and Virulence of Two Predominant Hospital-Acquired Methicillin-Resistant Staphylococcus aureus Clones and Clonal Methicillin-Susceptible S. aureus Isolates

The virulence of SCCmec type IV hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates belonging to the major sequence type 8 (ST8 [Lyon clone]) and to a minor upcoming clone, ST5, was compared with that of methicillin-susceptible S. aureus (MSSA) isolates of matching sequence types. In vitro adhesion to human airway epithelial cells (HAECs) as an indicator of dissemination and mortality in a murine sepsis model as an indicator of virulence were evaluated. Ten MRSA isolates and 8 MSSA isolates of ST8 and 8 MRSA isolates and 8 MSSA isolates of ST5 were characterized with respect to multilocus sequence type; agr, spa, and capsule typing; in vitro doubling time; toxin and adhesin gene profiles; and adherence to HAECs. Adherence was significantly lower in the MRSA ST5 group than in the ST8 groups. Infections with MRSA and MSSA isolates ST8 and ST5 were compared. No change in virulence related to the presence of SCCmec was observed, since ST8 but not ST5 caused a significantly lower mortality in its presence. Despite their similar genetic backgrounds, individual clonal MRSA and MSSA isolates were heterogeneous in adherence and virulence. No one of these specific virulence factors determined in vitro was related to mouse mortality. In conclusion, in a bacteremic model, mortality was dependent on the ST and was differentially modulated by SCCmec; within an ST, clonality was not associated with a homogenous outcome.

Incidence des Staphylococcus aureus résistants à la méticilline dans les infections cutanées de l’enfant survenues en milieu communautaire : étude rétrospective 2000–2005

A dramatic increase in the incidence of methicillin-resistant Staphylococcus aureus (MRSA) in community-onset skin infections has been reported over the last 10 years in the USA. The emergence of MRSA has been recently described in France. The aims of this study were to assess the incidence of MRSA in community-onset skin infections and to analyse the characteristics of MRSA skin infections in a French paediatric population.This is a retrospective study covering the period January 2000 to December 2005. Patients aged under 15 years with S. aureus isolated from skin and a clinical diagnosis of skin infection were included.One hundred and thirty-four children were included with a median age of 3.4 years. There were no significant differences in MRSA prevalence between the different years of the study. The overall prevalence of MRSA was 8.2% (n=11/134). None of the isolated strains presented an antimicrobial susceptibility profile suggestive of the ST80-type community-acquired MRSA described in France. Three MRSA strains were isolated from serious superantigen-mediated skin infections. The antimicrobial susceptibility and genetic profile (tst-positive agr2 MSRA) for one strain of S. aureus militated strongly in favour of an MRSA ST5 clone skin infection.In this study we found no evidence of epidemic spread of MRSA in community-onset childhood skin infections between 2000-2005. Nevertheless, we report three cases of serious MRSA-induced superantigen-associated skin infection. This argues in favour of the presence of virulent community MRSA clones in France.

Transmission of methicillin-resistant Staphylococcus aureus from food production animals to humans: a review.

Abstract International surveillance of antimicrobial use in food animal production shows that methicillin-resistant Staphylococcus aureus (MRSA), traditionally a human pathogen associated with hospitals, has emerged in the community and animals. Since 1961, MRSA has been causing human infections in hospitals worldwide and a vast majority of them were caused by five major epidemic clones. After 1990, other clones have emerged in the community, leading to infections in relatively young and healthy individuals. The origin of these clones is largely unknown, and extensive diversity among isolates exists. Companion animals have been indicated as a reservoir. However, most studies suggest that they are initially infected by humans and subsequently animals re-colonize humans. More recently, a new zoonotic reservoir in food production animals was found. This involves a specific clone, MRSA ST398, which spreads extensively in animals and is also found in retail meat. It poses a potential threat to public health, as people in contact with food production animals are at much higher risk of colonization. The most probable transmission route seems to be by (in)direct contact, as dust in stables was found positive for MRSA ST398. The role of MRSA ST398 as a food pathogen needs more research. To prevent colonization in humans, it is important to investigate transmission routes and transmission dynamics between animals, between animals and humans and between humans. Collaboration of human and veterinary epidemiologists and microbiologists is needed to identify the implications of this strain for public health and to develop cost-effective control strategies.