Marked Presence of Methicillin-Resistant Staphylococcus aureus in Wild Lagomorphs in Valencia, Spain

The appearance of methicillin-resistant strains of Staphylococcus aureus (MRSA) in several animal species (including rabbits) has set off alarms for their capacity to act as reservoirs for this bacterium. This is especially important in wild animals given its epidemiological implications. The objectives of this study were to identify and characterize S. aureus, specifically MRSA, strains in wild lagomorph high-density areas. Ten hares and 353 wild rabbits from 14 towns with a high rabbit density in the Valencian region (eastern Spanish coast) were sampled. Swabs from the nasal cavity, ears, perineum and lesions (when present) were taken for microbiological studies. The detection of different genes and antibiotic susceptibility studies were also carried out. Of all the animals, 41.3% were positive for S. aureus, of which 63.3% were MRSA. Ears were the anatomical location with more S. aureus and MRSA strains. The more frequently identified MLST type was ST1945 (97.1%, 136/140). The mecA gene was found only in one sample. The rest (n = 139) carried the mecC gene and were included in CC130, except one. Penicillin resistance was detected in 28 mec-negative isolates and, in one case, bacitracin resistance. mecA isolate presented resistance to enrofloxacin and tetracycline, and 10 mecC isolates also showed bacitracin resistance. No MRSA isolate was positive for genes chp, sea, tst and PVL. Two ST1945 isolates contained IEC type E (comprising genes scn and sak). mecA-isolate was positive for blaZ. Of the 28 MSSA strains showing resistance to penicillin, 22 carried the blaZ gene. These surprising results highlight the marked presence of MRSA strains in wild rabbits in high-density areas.

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Detection of mecA gene by latex agglutination and its molecular analysis by PCR in methicillin resistant staphylococcus aureus.

The Professional Medical Journal ◽

10.29309/tpmj/2020.27.07.3752 ◽

2020 ◽

Vol 27 (07) ◽

pp. 1363-1370

Author(s):

Aneela Khawaja ◽

Iffat Javed ◽

Sohaila Mushtaq ◽

Saeed Anwar ◽

Faiqa Arshad ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistant Staphylococcus Aureus ◽

Methicillin Resistance ◽

Meca Gene ◽

Latex Agglutination ◽

Medical Institute ◽

Cross Sectional ◽

Methicillin Resistant ◽

Agglutination Method ◽

Resistant Strains

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus. This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.

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Antibiotic Resistance and Virulence Gene Characteristics of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolated from Healthy Edible Marine Fish

International Journal of Microbiology ◽

10.1155/2020/9803903 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Justine Fri ◽

Henry A. Njom ◽

Collins N. Ateba ◽

Roland N. Ndip

Keyword(s):

Staphylococcus Aureus ◽

Antibiotic Resistance ◽

Marine Fish ◽

Control Strategies ◽

Virulence Gene ◽

Multidrug Resistant ◽

Methicillin Resistant ◽

Vancomycin Mics ◽

Resistant Strains ◽

Mrsa Strains

Thirty-three (33) isolates of methicillin-resistant Staphylococcus aureus (MRSA) from healthy edible marine fish harvested from two aquaculture settings and the Kariega estuary, South Africa, were characterised in this study. The phenotypic antimicrobial susceptibility profiles to 13 antibiotics were determined, and their antibiotic resistance determinants were assessed. A multiplex PCR was used to determine the epidemiological groups based on the type of SCCmec carriage followed by the detection of staphylococcal enterotoxin-encoding genes sea-sed and the Panton Valentine leucocidin gene (pvl). A high antibiotic resistance percentage (67–81%) was observed for Erythromycin, Ampicillin, Rifampicin, and Clindamycin, while maximum susceptibility to Chloramphenicol (100%), Imipenem (100%), and Ciprofloxacin (94%) was recorded. Nineteen (58%) of the MRSA strains had Vancomycin MICs of ≤2 μg/mL, 4 (12%) with MICs ranging from 4–8 μg/mL, and 10 (30%) with values ≥16 μg/mL. Overall, 27 (82%) isolates were multidrug-resistant (MDR) with Erythromycin-Ampicillin-Rifampicin-Clindamycin (E-AMP-RIP-CD) found to be the dominant antibiotic-resistance phenotype observed in 4 isolates. Resistance genes such as tetM, tetA, ermB, blaZ, and femA were detected in two or more resistant strains. A total of 19 (58%) MRSA strains possessed SCCmec types I, II, or III elements, characteristic of healthcare-associated MRSA (HA-MRSA), while 10 (30%) isolates displayed SCCmec type IVc, characteristic of community-associated MRSA (CA-MRSA). Six (18%) of the multidrug-resistant strains of MRSA were enterotoxigenic, harbouring the see, sea, or sec genes. A prevalence of 18% (6/33) was also recorded for the luk-PVL gene. The findings of this study showed that marine fish contained MDR-MRSA strains that harbour SCCmec types, characteristic of either HA-MRSA or CA-MRSA, but with a low prevalence of enterotoxin and pvl genes. Thus, there is a need for continuous monitoring and implementation of better control strategies within the food chain to minimise contamination of fish with MDR-MRSA and the ultimate spread of the bug.

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Findings of methicillin-resistant strains of Staphylococcus aureus in livestock

Czech Journal of Food Sciences ◽

10.17221/209/2009-cjfs ◽

2010 ◽

Vol 27 (Special Issue 2) ◽

pp. 36-41

Author(s):

Z. Šťástková ◽

S. Karpíšková ◽

R. Karpíšková

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistant Staphylococcus Aureus ◽

Meca Gene ◽

Potential Hazard ◽

The Czech Republic ◽

Methicillin Resistant ◽

Milk Samples ◽

Genes Encoding ◽

Resistant Strains ◽

Bulk Tank

The aim of our study was to determine the occurrence of methicillin resistant Staphylococcus aureus (MRSA) at dairy farms in the Czech Republic. Altogether 1061 samples from 95 farms were examined. The samples analysed were milk (individual and bulk tank milk samples), animal swabs and swabs from the farm environment. In total, 299 S. aureus isolates were obtained, of which 23 were MRSA. These MRSA isolates originated from three farms (13 isolates from farm A and 5 isolates from each of farms B and C). All MRSA isolates carried the mecA gene while none of them carried the genes for PVL, TSST-1 and exfoliatins. Only the isolates from goat farm C were positive for the genes encoding enterotoxins. By SCCmec typing, the strains were classified as community-associated MRSA carrying SCCmec IV or V. This study revealed that animals can be an important source of methicillin resistant staphylococci and represent a potential hazard of further spread.

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Missense Mutations in PBP2A Affecting Ceftaroline Susceptibility Detected in Epidemic Hospital-Acquired Methicillin-Resistant Staphylococcus aureus Clonotypes ST228 and ST247 in Western Switzerland Archived since 1998

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.04068-14 ◽

2015 ◽

Vol 59 (4) ◽

pp. 1922-1930 ◽

Cited By ~ 52

Author(s):

William L. Kelley ◽

Ambre Jousselin ◽

Christine Barras ◽

Emmanuelle Lelong ◽

Adriana Renzoni

Keyword(s):

Staphylococcus Aureus ◽

University Hospital ◽

Surveillance Program ◽

Missense Mutations ◽

Unknown Parameters ◽

Methicillin Resistant ◽

Content Type ◽

Resistant Strains ◽

Mrsa Strains

ABSTRACTThe development and maintenance of an arsenal of antibiotics is a major health care challenge. Ceftaroline is a new cephalosporin with activity against methicillin-resistantStaphylococcus aureus(MRSA); however, no reports concerning MRSA ceftaroline susceptibility have been reported in Switzerland. We tested thein vitroactivity of ceftaroline against an archived set of 60 MRSA strains from the University Hospital of Geneva collected from 1994 to 2003. Our results surprisingly revealed ceftaroline-resistant strains (MIC, >1 μg/ml in 40/60 strains; EUCAST breakpoints, susceptible [S], ≤1 μg/ml; resistant [R], >1 μg/ml) were present from 1998 to 2003. The detected resistant strains predominantly belonged to sequence type 228 (ST228) (South German clonotype) but also to ST247 (Iberian clonotype). A sequence analysis of these strains revealed missense mutations in the penicillin-binding protein 2A (PBP2A) allosteric domain (N146K or E239K and N146K-E150K-G246E). The majority of our ST228 PBP2A mutations (N146K or E150K) were distinct from ST228 PBP2A allosteric domain mutations (primarily E239K) recently described for MRSA strains collected in Thailand and Spain during the 2010 Assessing Worldwide Antimicrobial Resistance Evaluation (AWARE) global surveillance program. We also found that similar allosteric domain PBP2A mutations (N146K) correlated with ceftaroline resistance in an independent external ST228 MRSA set obtained from the nearby University Hospital of Lausanne, Lausanne, Switzerland, collected from 2003 to 2008. Thus, ceftaroline resistance was observed in our archived strains (including two examples of an MIC of 4 µg/ml for the Iberian ST247 clonotype with the triple mutation N146K/E150K/G246E), at least as far back as 1998, considerably predating the commercial introduction of ceftaroline. Our results reinforce the notion that unknown parameters can potentially exert selective pressure on PBP2A that can subsequently modulate ceftaroline resistance.

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Potensi Ekstrak Atuna racemosa sebagai Anti - Methicillin Resistant Staphylococcus aureus (MRSA)

Jurnal Sain Veteriner ◽

10.22146/jsv.34700 ◽

2017 ◽

Vol 35 (2) ◽

pp. 260

Author(s):

Siti Isrina Oktavia Salasia ◽

Novra Arya Sandi ◽

Fajar Budi Lestari ◽

Verda Farida ◽

Nurbani Aziz

Keyword(s):

Staphylococcus Aureus ◽

Meca Gene ◽

New Drugs ◽

Diffusion Test ◽

Methicillin Resistant ◽

Causative Agents ◽

Mrsa Infection ◽

Polymerase Chain ◽

Mrsa Strains ◽

Multiple Antibiotics

Staphylococcus aureus is one of the major causative agents of mastitis in animals and a variety of human diseases such as septicemia, endocarditis, arthritis dan osteomyelitis. Infection of Methicillin-resistant S. aureus (MRSA) has been widely reported and these strains are usually resistant to multiple antibiotics. The purpose of this study was to evaluate the potential of Atuna racemosa, as an alternative herbal medicine against MRSA infection. The MRSA strains were isolated from human and confirmed based on their resistant to various antibiotics and analyzing of the mecA gene by polymerase chain reaction (PCR). Atuna racemosa originated from Ambon, Maluku, Indonesia, were extracted using 70% ethanol. The activities of the Atuna racemosa extract against MRSA were performed by diffusion disc agar and dilussion agar tests. The results showed that Atuna racemosa extract has the barrier effect of MRSA growth at a concentration of 5% in the diffusion test and at a concentration of 7% in the dilution test. Atuna racemosa could be used as an alternative new drugs with dose of 0.07 g/ml (7%) against MRSA which is multi-resistant to many antibiotics.

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Phenotypic and Molecular Typing of Nosocomial Methicillin-Resistant Staphylococcus aureus Strains Susceptible to Gentamicin Isolated in France from 1995 to 1997

Journal of Clinical Microbiology ◽

10.1128/jcm.38.1.185-190.2000 ◽

2000 ◽

Vol 38 (1) ◽

pp. 185-190

Author(s):

Jacques-Olivier Galdbart ◽

Anne Morvan ◽

Nevine El Solh

Keyword(s):

Staphylococcus Aureus ◽

Molecular Typing ◽

Insertion Sequence ◽

Methicillin Resistant Staphylococcus Aureus ◽

Phage Typing ◽

Methicillin Resistant ◽

Before And After ◽

Resistant Strains ◽

Mrsa Strains

ABSTRACT Methicillin-resistant strains susceptible to gentamicin (Gm s MRSA) have emerged since 1993 in several French hospitals. To study whether particular clones have spread in various French cities and whether some clones are related to gentamicin-resistant (Gm r ) MRSA strains, various methods (antibiotyping, phage typing, determination of Sma I macrorestriction patterns before and after hybridization with IS 256 transposase and aacA-aphD probes) were used to compare 62 Gm s MRSA strains isolated from 1995 to 1997 in nine cities and 15 Gm r MRSA strains. Eighteen major Sma I genotypes were identified, of which 11 included only Gm s MRSA strains and 5 included only Gm r MRSA strains. Each of the Gm r MRSA strains contained 6 to 13 Sma I fragments hybridizing with the insertion sequence IS 256 , of which a single band also hybridized with the aacA-aphD gene. No such hybridizing sequences were detected in 60 of the 62 Gm s MRSA strains. Thus, the divergence between Gm r and Gm s MRSA strains is revealed, not only by their distributions in distinct Sma I genotypes but also by the differences in hybridization patterns. Two of the 62 Gm s MRSA strains had the uncommon feature of carrying several Sma I bands hybridizing with IS 256 , suggesting that they are possibly related to the Gm r MRSA strains grouped in the same Sma I genotype. Five of the 11 Sma I genotypes including only Gm s MRSA strains contained strains from diverse cities, isolated during different years and with different antibiograms, suggesting that some clones have spread beyond their cities of origin and persisted.

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Livestock-Associated Methicillin-Resistant Staphylococcus aureus (MRSA) in Purulent Subcutaneous Lesions of Farm Rabbits

Foods ◽

10.3390/foods9040439 ◽

2020 ◽

Vol 9 (4) ◽

pp. 439 ◽

Cited By ~ 1

Author(s):

Vanessa Silva ◽

Telma de Sousa ◽

Paula Gómez ◽

Carolina Sabença ◽

Madalena Vieira-Pinto ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Agents ◽

Methicillin Resistant Staphylococcus Aureus ◽

Multidrug Resistant ◽

Diffusion Method ◽

Spa Typing ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Resistance To Penicillin ◽

Mrsa Strains

Methicillin-resistant Staphylococcus aureus (MRSA) are one of the main pathogens associated with purulent infections. MRSA clonal complex 97 (CC97) has been identified in a wide diversity of livestock animals. Therefore, we aimed to investigate the antibiotic resistance profiles of MRSA strains isolated from purulent lesions of food-producing rabbits. Samples from purulent lesions of 66 rabbits were collected in a slaughterhouse in Portugal. Samples were seeded onto ORSAB plates with 2 mg/L of oxacillin for MRSA isolation. Susceptibility to antibiotics was tested by the disk diffusion method against 14 antimicrobial agents. The presence of resistance genes, virulence factors and the immune evasion cluster (IEC) system was studied by polymerase chain reaction. All isolates were characterized by multilocus sequence typing (MLST), agr and spa typing. From the 66 samples analyzed, 16 (24.2%) MRSA were detected. All strains were classified as multidrug-resistant as they were resistant to at least three classes of antibiotics. All isolates showed resistance to penicillin, erythromycin and clindamycin. Seven isolates were resistant to gentamicin and harbored the aac(6′)-Ie-aph (2″)-Ia gene. Resistance to tetracycline was detected in 10 isolates harboring the tet(K) gene. The IEC genes were detected in three isolates. MRSA strains belonged to CC97, CC1, CC5, CC15 or CC22. The isolates were assigned to six different spa types. In this study we found a moderate prevalence of multidrug-resistant MRSA strains in food-producing rabbits. This may represent concern for food safety and public health, since cross-contamination may occur, leading to the spread of MRSA and, eventually, the possibility of ingestion of contaminated meat.

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Methicillin-resistant Staphylococcus spp. isolated from curd cheese “requeijão” and “especialidade láctea type requeijão” sold in Brazil

Ciência Rural ◽

10.1590/0103-8478cr20170008 ◽

2017 ◽

Vol 47 (7) ◽

Author(s):

Jamile de Oliveira Hachiya ◽

Gabriel Augusto Marques Rossi ◽

Laryssa Freitas Ribeiro ◽

Rafael Akira Sato ◽

Higor Oliveira Silva ◽

...

Keyword(s):

Dairy Products ◽

Meca Gene ◽

Multidrug Resistant ◽

Human Medicine ◽

Methicillin Resistant ◽

Heat Treated ◽

Antimicrobial Resistance Profile ◽

Resistance To Penicillin ◽

Resistant Strains ◽

Staphylococcus Spp

ABSTRACT: This study focused on counting Staphylococcus spp. in curd cheeses “requeijão” and “especialidade láctea type requeijão” sold in Brazil, assessing the presence of mecA gene in obtained isolates and establishing antimicrobial resistance profile of the mecA gene positive isolates. To this, a set of 200 samples of these dairy products were evaluated. Low counts of Staphylococcus spp. were observed in these dairy products. All the isolates were determined as coagulase-negative strains using coagulase test and PCR. However, two isolates (3.70%) were carriers of mecA gene and they can be considered as risk for public health. These isolates presented resistance to penicillin, oxacillin and erythromycin. In conclusion, low counts of Staphylococcus were detected in curd cheese “requeijão” and “especialidade láctea type requeijão” sold in Brazil. However, coagulase-negative methicillin-resistant Staphylococcus spp. was detected in these dairy products. This is the first report of the detection of methicillin-resistant Staphylococcus spp. in heat-treated dairy products in Brazil. Results served as a warning to public sanitary authorities to control multidrug-resistant strains in veterinary and human medicine.

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Staphylococcus aureus PBP4 Is Essential for β-Lactam Resistance in Community-Acquired Methicillin-Resistant Strains

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00049-08 ◽

2008 ◽

Vol 52 (11) ◽

pp. 3955-3966 ◽

Cited By ~ 105

Author(s):

Guido Memmi ◽

Sergio R. Filipe ◽

Mariana G. Pinho ◽

Zhibiao Fu ◽

Ambrose Cheung

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistance ◽

Healthy Individuals ◽

Penicillin Binding Protein ◽

Methicillin Resistant ◽

Fold Reduction ◽

Resistant Strains ◽

Mrsa Strains ◽

Hospital Acquired ◽

Major Target

ABSTRACT Recent cases of infections caused by community-acquired methicillin-resistant Staphylococcus aureus (MRSA) (CA-MRSA) strains in healthy individuals have raised concerns worldwide. CA-MRSA strains differ from hospital-acquired MRSAs by virtue of their genomic background and increased virulence in animal models. Here, we show that in two common CA-MRSA isolates, USA300 and MW2 (USA400), a loss of penicillin binding protein 4 (PBP4) is sufficient to cause a 16-fold reduction in oxacillin and nafcillin resistance, thus demonstrating that mecA, encoding PBP2A, is not the sole determinant of methicillin resistance in CA-MRSA. The loss of PBP4 was also found to severely affect the transcription of PBP2 in cells after challenge with oxacillin, thus leading to a significant decrease in peptidoglycan cross-linking. Autolysis, which is commonly associated with the killing mechanism of penicillin and β-lactams, does not play a role in the reduced resistance phenotype associated with the loss of PBP4. We also showed that cefoxitin, a semisynthetic β-lactam that binds irreversibly to PBP4, is synergistic with oxacillin in killing CA-MRSA strains, including clinical CA-MRSA isolates. Thus, PBP4 represents a major target for drug rediscovery against CA-MRSA, and a combination of cefoxitin and synthetic penicillins may be an effective therapy for CA-MRSA infections.

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Phenotypic and Genotypic Characterization of Nosocomial Staphylococcus aureus Isolates from Trauma Patients

Journal of Clinical Microbiology ◽

10.1128/jcm.36.2.414-420.1998 ◽

1998 ◽

Vol 36 (2) ◽

pp. 414-420 ◽

Cited By ~ 32

Author(s):

T. Na’was ◽

A. Hawwari ◽

E. Hendrix ◽

J. Hebden ◽

R. Edelman ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Trauma Patients ◽

Resistant Genotype ◽

Methicillin Resistant ◽

University Of Maryland ◽

Single Clone ◽

Resistant Strains ◽

Mrsa Strains ◽

The University

Staphylococcus aureus is a major cause of nosocomial infections. During the period from March 1992 to March 1994, the patients admitted to the intensive care unit of the University of Maryland Shock Trauma Center were monitored for the development ofS. aureus infections. Among the 776 patients eligible for the study, 60 (7.7%) patients developed 65 incidents of nosocomialS. aureus infections. Of the clinical isolates, 43.1% possessed a polysaccharide type 5 capsule, 44.6% possessed a type 8 capsule, and the remaining 12.3% had capsules that were not typed by the type 5 or type 8 antibodies. Six antibiogram types were noted among the infection-related isolates, with the majority of the types being resistant only to penicillin and ampicillin. It was noted that the majority of cases of pneumonia were caused by relatively susceptible strains, while resistant strains were isolated from patients with bacteremia and other infections. Only 16 (6.3%) of the isolates were found to be methicillin-resistant S. aureus (MRSA). DNA fingerprinting by pulsed-field gel electrophoresis showed 36 different patterns, with characteristic patterns being found for MRSA strains and the strains with different capsular types. Clonal relationships were established, and the origins of the infection-related isolates in each patient were determined. We conclude that (i) nosocomial infection-related isolates from the shock trauma patients did not belong to a single clone, although the predominance of a methicillin-resistant genotype was noted, (ii) most infection-relatedS. aureus isolates were relatively susceptible to antibiotics, but a MRSA strain was endemic, and (iii) for practical purposes, the combination of the results of capsular and antibiogram typing can be used as a useful epidemiological marker.

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