Methicillin-resistant Staphylococcus Aureus Genotypes Research Articles

Clinical and molecular characteristics of methicillin-resistant Staphylococcus aureus in bone and joint infection among children

ObjectiveTo investigate the characteristics of Methicillin-Resistant Staphylococcus aureus (MRSA) in bone and joint infection (BJI) among children.MethodsA total of 338 patients diagnosed with BJI from 2013 to 2022 in Children’s Hospital of Fudan University were enrolled. Demographic information, microbiology culture results and laboratory findings, including white blood counts (WBC), C-reactive protein (CRP), procalcitonin (PCT), interleukin-6 (IL-6), and erythrocyte sedimentation rate (ESR) were collected and analyzed. MRSA was confirmed by antimicrobial susceptibility testing. Other MRSA-caused infections were randomly selected for comparison. Twenty-three virulence and antimicrobial resistance (AMR) genes were screened for MRSA strains. Multilocus sequence typing (MLST) and Staphylococcal protein A (spa) typing were performed using PCR amplification and sequencing.ResultsOf the identified pathogens in BJI, MRSA accounted for 21.0% (47/224). Patients with BJI had high levels of initial CRP, white blood cell count (WBC) and IL-6. ST59 (43.9%) and t437 (37.6%) were the main MRSA subtypes isolated from the children. The major genotypes in BJI were ST59-t437 (29.8%) and ST22-t309 (14.9%), with high carriage of hemolysins including hla (94.4–100%), hlb (66.2–93.3%), and hld (100%). Notably, Panton–Valentine leukocidin (pvl) had a high prevalence (53.3%) in ST22-t309-MRSA. Other virulence genes including tst, seg and sei were more commonly detected in ST22-t309-MRSA (40.0–46.7%) than in ST59-t437-MRSA (4.2–9.9%). High-carriage AMR genes in MRSA included aph(3ʹ)/III (66.7–80%), ermB (57.5–73.3%) and ermC (66.7–78.9%). MRSA presented high-resistance to erythromycin (52.0–100%) and clindamycin (48.0–92.5%), different genotypes displayed variation in their susceptibilities to antibiotics.ConclusionsThe major MRSA genotype in BJI was ST59-t437, followed by ST22-t309, with a higher prevalence of the pvl gene. Continuous surveillance of pvl-positive ST22-t309-MRSA in pediatric BJI infections is thus required.

Bacterial Genotype, Carrier Risk Factors, and an Antimicrobial Stewardship Approach Relevant to Methicillin-resistant Staphylococcus aureus Prevalence in a Population of Macaques Housed in a Research Facility.

Methicillin-resistant Staphylococcus aureus (MRSA) remains a significant problem for human and animal health and can negatively affect the health status of macaques and other nonhuman primates (NHP) in research colonies. However, few publications provide guidance on the prevalence, genotype, or risk factors for macaques with MRSA and even fewer on how to effectively respond to MRSA once identified in a population. After having a clinical case of MRSA in a rhesus macaque, we sought to determine the MRSA carrier prevalence, risk factors, and genotypes of MRSA in a population of research NHPs. Over a 6-wk period in 2015, we collected nasal swabs from 298 NHPs. MRSA was isolated from 28% (n = 83). We then reviewed each macaque's medical record for a variety of variables including animal housing room, sex, age, number of antibiotic courses, number of surgical interventions, and SIV status. Analysis of these data suggests that MRSA carriage is associated with the room location, age of the animal, SIV status, and the number of antibiotic courses. We used multilocus sequence typing and spa typing on a subset of MRSA and MSSA isolates to determine whether the MRSA present in NHPs was comparable with common human strains. Two MRSA sequence types were predominant: ST188 and a novel MRSA genotype, neither of which is a common human isolate in the United States. We subsequently implemented antimicrobial stewardship practices (significantly reducing antimicrobial use) and then resampled the colony in 2018 and found that MRSA carriage had fallen to 9% (26/285). These data suggest that, as in humans, macaques may have a high carrier status of MRSA despite low clinically apparent disease. Implementing strategic antimicrobial stewardship practices resulted in a marked reduction in MRSA carriage in the NHP colony, highlighting the importance of limiting antimicrobial use when possible.

Diversity and Dissemination of Methicillin-Resistant Staphylococcus aureus (MRSA) Genotypes in Southeast Asia.

Methicillin-resistant Staphylococcus aureus (MRSA) is a successful pathogen that has achieved global dissemination, with high prevalence rates in Southeast Asia. A huge diversity of clones has been reported in this region, with MRSA ST239 being the most successful lineage. Nonetheless, description of MRSA genotypes circulating in the Southeast Asia region has, until now, remained poorly compiled. In this review, we aim to provide a better understanding of the molecular epidemiology and distribution of MRSA clones in 11 Southeast Asian countries: Singapore, Malaysia, Thailand, Vietnam, Cambodia, Lao People's Democratic Republic (PDR), Myanmar, Philippines, Indonesia, Brunei Darussalam, and Timor-Leste. Notably, while archaic multidrug-resistant hospital-associated (HA) MRSAs, such as the ST239-III and ST241-III, were prominent in the region during earlier observations, these were then largely replaced by the more antibiotic-susceptible community-acquired (CA) MRSAs, such as ST22-IV and PVL-positive ST30-IV, in recent years after the turn of the century. Nonetheless, reports of livestock-associated (LA) MRSAs remain few in the region.

Characterization of Methicillin-Resistant Staphylococcus aureus Isolates from Periprosthetic Joint Infections.

Periprosthetic joint infection (PJI) is a troublesome clinical issue in total joint arthroplasty (TJA). Although methicillin-resistant Staphylococcus aureus (MRSA) is considered to be the most serious pathogen in PJIs, little is known about the genotypic and phenotypic characteristics of MRSA clones isolated from PJI patients. A total of 36 MRSA isolates from PJI patients were collected at the Chang-Gung Memorial Hospital in Taiwan from May 2016 to October 2019. All MRSA isolates were subjected to genome typing. The prevalence of Panton–Valentine leucocidin (PVL), the antibiotic susceptibility profile, and the biofilm formation ability were compared among different MRSA genogroups. Additionally, demographics and clinical manifestations of patients infected with different MRSA genogroups were investigated. Eight sequence types (STs) were identified among 36 isolated from PJIs. According to the incidence of MRSA genotypes in PJIs, in this study, we divided them into four groups, including ST8 (n = 10), ST59 (n = 8), ST239 (n = 11), and other STs (n = 7). For the antibiotic susceptibility testing, we found that all MRSA isolates in the ST239 group were highly resistant to ciprofloxacin, gentamicin trimethoprim-sulfamethoxazole, and levofloxacin. Additionally, ST239 MRSA also had a higher ability to form biofilm than other groups. Importantly, patients with ST239 infection typically had a fever and exhibited higher levels of inflammatory markers, including C-reactive protein (CRP) and white blood cell count (WBC). Epidemiological investigations revealed that knee PJIs were mainly attributed to infection with ST59 MRSA and increasing trends for infection with ST8 and other ST types of MRSAs in PJI patients were observed from 2016 to 2019. The identification of MRSA genotypes in PJIs may be helpful for the management of PJIs.

Distribution of sasX, qacA/B and mupA genes and determination of genetic relatedness of methicillin-resistant Staphylococcus aureus among clinical isolates and nasal swab samples from the same patients in a hospital in Malaysia.

This study determined the distribution of sasX, qacA/B and mupA genes from methicillin-resistant Staphylococcus aureus (MRSA) isolated from clinical samples and nasal swab samples of the same patients and analysed their genetic relatedness. Polymerase chain reaction was used to detect the presence of sasX, qacA/B and mupA genes from 47 paired MRSA isolates. A paired isolate was defined as one nasal swab (colonising) isolate and clinical isolate that caused infection in the same patient. 22 selected paired isolates were subjected to multilocus sequence typing (MLST). The genetic relatedness among the isolates and association between the putative genes with epidemic sequence types (STs) were investigated. 7 (14.9%, n = 14) paired isolates were positive for the sasX gene. qacA/B genes were positive in 7.4% (n = 7) of the isolates, from three paired isolates and one clinical isolate whose paired colonising isolate was negative. The paired sample of three patients were positive for both genes. The mupA gene was not detected in all the isolates. MLST revealed two epidemic STs, ST22 and ST239, and a novel ST4649. sasX and qacA/B genes were found in ST239 in 29.5% (n = 13) and 13.6% (n = 6) of cases, respectively. Gene co-existence occurred in 13.6% (n = 6) of MRSA ST239 and 2.3% (n = 1) of MRSA ST4649. sasX and qacA/B genes were present in the MRSA isolates, while the mupA gene was undetected. ST22 and ST239 were the major MRSA clones. The circulating MRSA genotypes conferred different virulence and resistance determinants in our healthcare settings.

Genomic portrait of community-associated methicillin-resistant Staphylococcus aureus ST772-SCCmec V lineage from India

BackgroundSignificant changes in the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) were recognised with the emergence of community-associated methicillin-resistant Staphylococcus aureus. However, studies on the molecular epidemiology and the genomic investigation of MRSA are limited in India. AimThe study was aims to understand the molecular epidemiology of MRSA causing bloodstream infection and also to investigate the origin and evolution of ST772 S. aureus isolated from India. MethodsA total of 233 non-repetitive MRSA isolates were screened for the presence staphylococcal cassette chromosome (SCCmec) types, multi-locus sequence types (MLST) and staphylococcal protein A (spa) types. Whole genome sequence data of ST772-SCCmec V (n = 32) isolates were generated and comparative analysis was performed. ResultsMLST analysis revealed ten different clonal complexes and three singletons. ST772 (27%), ST22 (19%) and ST239 (16%) were the predominant MRSA genotypes in causing bloodstream infection. The spa types were highly diverse. Phylogenetic analysis revealed that nearly three-fourth of the Indian STT72-SCCmec V isolates belongs to dominant (ST772-A2) and emerging subgroups (ST772-A3). A pattern of increasing antimicrobial resistance was noticed in the dominant and emerging subgroups. An integrated resistance plasmid encoding resistance clusters for beta-lactam (blaZ), macrolides (mphC, msrA), and aminoglycoside resistance (aphA-III, sat-4, aadE) was identified in all isolates, except four basal strains. ST772-SCCmec V was emerged on the Indian subcontinent in 1964 and diverged into a dominant subgroup in 1991. Furthermore, the expansion is likely to be associated with the acqusition of mobile genetic elements such as integrated resistance plasmid and SCCmec V (5C2) as well as the fixation of double serine mutation (S84L, S80Y) in the quinolone resistance determining region. ConclusionsST772 S. aureus have consistent virulence and resistance determinants which may results in successful survival in both community and hospital settings.

Molecular characterization of methicillin-resistant and -susceptible Staphylococcus aureus recovered from hospital personnel

Introduction. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major causes of hospital-acquired infections. Over the past two decades MRSA has become 'epidemic' in many hospitals worldwide. However, little is known about the genetic background of S. aureus recovered from hospital personnel in China.Hypothesis/Gap Statement. The diversity of S. aureus genotypes warrants further surveillance and genomic studies to better understand the relatedness of these bacteria to those recovered from patients and the community.Aim. The aim of this study was to determine the genetic diversity of MRSA and methicillin-susceptible S. aureus (MSSA) recovered from hospital personnel in Tianjin, North China.Methodology. Three hundred and sixty-eight hand or nasal swabs were collected from 276 hospital personnel in 4 tertiary hospitals in Tianjin, North China between November 2017 and March 2019. In total, 535 Gram-positive bacteria were isolated, of which 59 were identified as S. aureus. Staphylococcal cassette chromosome mec (SCCmec) typing, multi-locus sequence typing (MLST) and spa typing were performed to determine the molecular characteristics of S. aureus.Results. Thirty-one out of 276 (11 %) hospital personnel were S. aureus carriers, whereas 11/276 (4 %) carried MRSA. Fifty out of 59 (85 %) S. aureus isolates were resistant or intermediately resistant to erythromycin. The dominant genotypes of MRSA recovered from hospital personnel were ST398-t034-SCCmecIV/V and ST630-t084/t2196, whereas the major genotypes of MSSA included ST15-t078/t084/t346/t796/t8862/t8945/t11653 and ST398-t189/t034/t078/t084/t14014.Conclusion. Although the predominant genotypes of MRSA recovered from hospital personnel in this study were different from the main genotypes that have previously been reported to cause infections in Tianjin and in other geographical areas of China, the MRSA ST398-t034 genotype has previously been reported to be associated with livestock globally. The dominant MSSA genotypes recovered from hospital personnel were consistent with the those previously reported to have been recovered from the clinic.

The occurrence and distribution of livestock-associated methicillin-resistant Staphylococcus aureus ST398 on German dairy farms

The objective of this study was to investigate the occurrence and distribution of methicillin-resistant Staphylococcus aureus (MRSA) on 20 German dairy farms. Farms were selected based on previous MRSA reports from phenotypic susceptibility testing of mastitis pathogens. Samples were collected from predefined groups of cows, young stock, farm personnel, and the environment. A high MRSA-positive test rate was detected in swab samples from milk-fed calves (22.7%; 46/203). In postweaning calves, the MRSA-positive test rate was 9.1% (17/187). From prefresh heifers, both nasal swabs and udder cleft swabs were collected if possible. Including both sample types, the MRSA-positive test rate in prefresh heifers was 13.0% (26/200). The positive test rate was 8.9% (17/191) in nasal swabs and 6.5% (11/170) in udder cleft swabs. In quarter milk samples (QMS), the MRSA-positive test rate was 2.9% (67/2347), and on cow level, 7.9% (47/597) of the dairy cows were affected. Among all cows included in this study, the geometric mean of somatic cell counts was higher in QMS that carried MRSA (345,000 cells/mL) in comparison to all QMS (114,000 cells/mL). No differences in parity or the affected mammary quarter position on the udder were observed among the 47 infected cows. Methicillin-resistant S. aureus was also detected in boot swab samples (dust), teat liners, and in suckers from automatic calf feeders. All isolates belonged to livestock-associated sequence type 398 and most common staphylococcal protein A (spa)-types were t011 and t034. Most isolates harbored the staphylococcal cassette chromosome mec (SCCmec)-type V, with the exception of some isolates with SCCmec-type IVa on 1 farm. Similar MRSA genotypes in samples from humans and dairy cows underline the possible zoonotic and reverse-zoonotic transmission of livestock-associated MRSA strains from dairy farms. Similar MRSA genotypes in pig and cattle barns were detected on only 1 of 5 farms that kept both cattle and pigs. Similar MRSA spa-types were detected in samples from different sources (dairy cows, young stock, environment, and humans), suggesting a possible contagious transmission on some of the farms. Sporadically, up to 3 different MRSA spa-types were detected in QMS from the respective farms. On MRSA-affected farms, improper milking hygiene procedures and elevated bulk-tank milk somatic cell counts (>250,000 cells/mL) were observed. The occurrence of livestock-associated MRSA ST398 in different samples from dairy farms, and especially in young calves, should be considered for future MRSA-monitoring programs and biosecurity guidelines.

Characterization of different virulent factors in methicillin-resistant Staphylococcus aureus isolates recovered from Iraqis and Syrian refugees in Duhok city, Iraq.

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious public health problem. There is limited information regarding the genetics of MRSA strains among the native Iraqi and incoming Syrian refugee communities. We aimed to characterize the genotypes and different virulence factors of MRSA in strains isolated from these two communities. Frozen MRSA strains (125) isolated from the native Iraqi and Syrian refugee communities were used in this study. PCR (singleplex and multiplex) and agr typing was used for the genotypic analysis of different virulence genes. We tested for the presence of virulence genes including pvl, arcA, tst, lukE/lukD, hla, hlb, eta, etb and agr. Prevalence of arcA MRSA in the Iraqi community (56.58%) was significantly higher (p = 0.008) than that in the Syrian refugee community (32.66%). Prevalence of lukE-lukD was also significantly higher (p = 0.001) in the Iraqi (82.89%) compared to that in the Syrian refugee community (57.14%). Further, prevalence of hla MRSA in the Iraqi community was (93.4%) and in the Syrian refugee community was (71.4%); (p = 0.0008). No significant differences were observed in the prevalence of pvl, tst, eta, etb and hlb. The most dominant agr types in both Iraqi (76.1% and 10.5%) and Syrian refugee (44.9% and 18.37%) communities were I and III. To sum up, no significant differences were observed between the groups for a majority of virulence factors. This is the first investigation of MRSA genotypes and virulence in both these communities. These results could be useful for further studies that assess the genetic relatedness of strains in the region for epidemiological and monitoring purposes, which would be crucial to limiting the spread of MRSA.

Epidemiology of methicillin-resistant Staphylococcus aureus in a Japanese neonatal intensive care unit.

There have been few reports on the genetic structure of the current population of methicillin-resistant Staphylococcus aureus (MRSA) from neonatal intensive care units (NICUs) in Japan. In the present study we conducted a molecular epidemiological analysis based on whole genome sequencing against MRSA strains in a Japanese NICU. We performed genotyping by whole genome sequencing, polymerase chain reaction-based typing of Staphylococcal cassette chromosome mec (SCCmec) and polymerase chain reaction-based open-reading frame typing against 57 MRSA strains from fecal or nasal specimens from NICU patients in Juntendo University Shizuoka Hospital in 2013-2014. Forty-nine MRSA strains (86.0%) exhibited a clonal complex (CC) 1, and were divided into three sequence types (STs): ST2725 (n = 25), ST2764 (n = 21), and ST1 (n = 3). All CC1 MRSA strains had SCCmec IVa, and were resistant to new quinolones, which are limited in pediatric use, suggesting that these strains were derived from adult MRSA clones. Single nucleotide polymorphism differences of both ≤10 and >100 nucleotides were observed by pairwise, single nucleotide polymorphism analysis among ST2725 and ST2764 MRSA strains, respectively. Seven ST8 MRSA strains (12.2%) were isolated, and no strain exhibiting the Japanese hospital-associated MRSA genotype (ST5/SCCmec II) was isolated in this study. Our molecular epidemiological analysis suggested that ST2725 and ST2764 MRSA strains had genetic diversity that could not be explained only by a recent transmission event in the NICU. These MRSA clones might be disseminated in other Japanese hospital facilities as new endemic clones. Our results are expected to contribute to the improvement of infection control measures of MRSA in NICUs.

Molecular analysis of methicillin-resistant Staphylococcus aureus isolates from four teaching hospitals in Iran: the emergence of novel MRSA clones

BackgroundThe global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan.MethodsA total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antimicrobial resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multilocus sequence typing and SCCmec typing.ResultsOf 146 Staphylococcus aureus isolates, 24 (16.4%) isolates were identified as MRSA strains. According to antimicrobial susceptibility testing the highest resistance rates were seen for tetracycline, erythromycin, ciprofloxacin and gentamicin. All of Staphylococcus aureus isolates were susceptible to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCCmec types were obtained among MRSA strains including 16 (66.7%) SCCmec type V, 3 (12.5%) SCCmec type III and 5 (20.8%) SCCmec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%), cna (25.0%), fnbB (16.6%), ebps 5 (20.8%), and the fnbA, bbp and clfA genes were not detected in any MRSA isolate. MLST analysis revealed 11 sequence types among MRSA isolates as follows: ST239, ST291, ST22, ST861, ST889, ST8, ST59, ST343, ST772, ST6 and ST1465. Also seven MLST-based clonal complexes (CCs) were identified among MRSA strains including: CC8, CC7, CC398, CC59, CC22, CC1 and CC5.ConclusionsA relatively high diversity was found in MRSA genotypes in Kashan and Isfahan hospitals, and seven clonal complexes were identified. Pandemic MRSA clones including CC8 and CC22 were the most prevalent clones and the novel ST types including ST1465, ST861, ST 889 and ST772 are reported for the first time in Iran in the present study. In addition the high prevalence of MSCRAMMs genes in MRSA isolates demonstrates the high potential of these strains for pathogenicity.

Antimicrobial Resistance and Molecular Characteristics of Methicillin-resistant Staphylococcus aureus Isolates from Children Patients in Iran.

IntroductionMethicillin-resistant Staphylococcus aureus (MRSA) causes high rates of mortality and a substantial burden to health systems worldwide. Here, we investigated the antimicrobial susceptibility and molecular characteristics of MRSA isolated from children referred to Children’s Medical Center in Tehran.Materials and methodsA total of 98 MRSA isolates were collected from children. Antimicrobial resistance patterns were determined using the disk diffusion and E-test methods. The presence of biofilm encoding genes and the pvl gene were determined by PCR. We used the microtiter plate method to assess the ability of biofilm formation. The MRSA isolates were further analyzed using PFGE and SCCmec typing.ResultsAntibiotic susceptibility testing showed that the highest and the lowest antibiotic resistance percentage were related to erythromycin (62%) and minocycline (10%), respectively. Overall, 63% of MRSA isolates were biofilm producers. Resistance to two antibiotics such as erythromycin (72% vs 28%, P=0.01) and clindamycin (71% vs 29%, P=0.04) was higher among biofilm producers than non-biofilm producers. All strains had biofilm-forming genes and the prevalence of pvl gene was 41%. Most MRSA isolates belonged to SCCmec IVa (75%) and SCCmec III (18%). In PFGE technique, 5 common types and 2 single types were identified; Common type 1 with 37 isolates was dominant clone.ConclusionWe thus report preliminary data on the prevalence and distribution of MRSA genotypes in Tehran Children’s Hospital. These findings characterize the MRSA colonization dynamics in child patients in Iran and may aid the design of strategies to prevent MRSA infection and dissemination.

Emerging variants of methicillin-resistant Staphylococcus aureus genotypes in Kuwait hospitals.

BackgroundFrequent changes in the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) occurring worldwide demand regular surveillance to study their composition and distribution in healthcare facilities. We investigated the genotypic characteristics of MRSA obtained in Kuwait hospitals to better understand their clonal distribution.Materials and methodsA total of 1,327 MRSA isolates obtained from clinical samples in 13 Kuwait hospitals from 1 January to 31 December 2016 were investigated using antibiogram, SCCmec typing, spa typing and DNA microarray.ResultsThe isolates belonged to six SCCmec types with the majority belonging to type IV (658; 49.5%) and type V (355; 26.7%). Two hundred and sixty-one spa types were identified with spa types t688, t304, t860, t127, t044, t311, t002, t223, t267, t019, t3841, t005, t084, t852, and t657 constituting 51.0% (n = 677) of the isolates. Among the 1,327 MRSA isolates, 102 (7.68%) isolates were identified as novel variants of internationally recognized MRSA clones. These 102 isolates were investigated further and belonged to 14 clonal complexes (CCs) with CC361 (32; 32.3%), CC30 (15; 14.7%), CC22 (13; 12.7%) and CC1 (11, 10.7%) as the dominant CCs. Eighty-one (79.4%) of the novel isolates harbored SCCmec IV or V+fusC composite genetic elements. Four isolates (3.9%) harbored unusual combinations of ccr and mec complexes comprising of CC6-MRSA [IV+fusC+ccrC], CC97-MRSA [V/VT+fusC+ccrAB2], CC121-MRSA [V/VT+fusC+ccrB4] and CC1-MRSA-pseudoSCCmec [class B mec+fusc+ccrAB1]. Forty-six (45.1%) of these isolates were positive for PVL and 89 (87.2%) were resistant to fusidic acid mediated by fusC.ConclusionsThe study showed the emergence of novel variants of previously recognized MRSA genotypes with unusual genetic characteristics including high prevalence of PVL and fusidic acid resistance in Kuwait hospitals. This has added to the dynamic lists of known variations in MRSA genomes which can impose serious challenges for infection control and treatment of MRSA infections.

In Vitro Susceptibility of Ciprofloxacin-Resistant Methicillin-Resistant Staphylococcus aureus to Ototopical Therapy.

Objective Despite the rising incidence of methicillin-resistant Staphylococcus aureus (MRSA) otologic infections, choice of treatment remains controversial. Only fluoroquinolone-containing ototopical preparations are approved by the US Food and Drug Administration for middle ear application. Furthermore, American Academy of Otolaryngology-Head and Neck Surgery Foundation guidelines advocate ototopical monotherapy for both tympanostomy tube otorrhea and acute otitis externa. Unfortunately, MRSA may be ciprofloxacin resistant. This causes confusion regarding antibiotic selection, because susceptibility profiles reflect a minimum inhibitory concentration (MIC), referenced against systemic, not ototopical, drug delivery dosing. The goal of this study is to determine the ciprofloxacin MIC for ciprofloxacin-resistant MRSA isolates from otologic infections and compare that value to the expected drug concentration achieved by fluoroquinolone ear drops and determine MRSA genotype for each isolate. Study Design In vitro assay with retrospective medical record review. Setting Tertiary care university hospital. Subjects and Methods Thirty otologically sourced ciprofloxacin-resistant MRSA isolates collected from adult and pediatric patients. MICs were calculated by broth dilution method. Isolates underwent multilocus sequence typing and polymerase chain reaction for arcA and Panton-Valentine leukocidin to establish the genotype. Results MICs ranged from 16 to 1025 µg/mL. There was a relationship between MIC and genotype; of the 7 isolates with an MIC value greater than 512 µg/mL, 6 were sequence type (ST)8. Conclusion These findings support the practice of ototopical monotherapy for patients with uncomplicated ciprofloxacin-resistant MRSA otitis externa. However, they raise concern that ototopical therapy may not be adequate to treat highly resistant strains of MRSA infecting the middle ear space.

Diversity of Multidrug Efflux Genes and Phenotypic Evaluation of the In vitro Resistance Dynamics of Clinical Staphylococcus Aureus Isolates Using Methicillin; a Model β-lactam.

Objectives:Methicillin-resistant Staphylococcus aureus (MRSA) across the world often leave clinicians with little or no choice of treatment options. The multi-drug efflux (MDE) genes are bacterial survival mechanisms responsible for the pumping out of antibiotics and other biocides from the cytoplasm. Whilst effort is being made in the development of antibiotic adjuvants such as efflux pumps inhibitors, information is needed on the diversity of these MDEs in the circulating S. aureus and on the growth dynamics of the clinical isolates in response to antibiotics is not regularly examined.Methods:Here, we evaluated the diversity of MDEs in cinical S. aureus recovered in a tertiary academic hospital, Pretoria, South African hospital using PCR and also employed visual minimum inhibitory concentration and quantitative analysis of spectrophometric measurements of bacterial growth in the presence of a model β lactam antibiotic (methicillin), to phenotypically elucidate the resistance pattern of these isolates in response to methicillin.Results:Three major distribution patterns of MDEs were observed in the clinical isolates evaluated. Moreover, norA, nor B and tet38 were present in 98.9% of the isolates while other MDE were present in different proportions ranging from 40 to 98.6% of the isolates. In addition, S. aureus isolates, be it of MRSA or MSSA genotype did not habour the same set of MDEs despite being recovered from the same hospital setting. Finally, we showed that MSSA displayed phenotypic resistance to methicilllin despite the non-detection of the mecA resistance gene.Conclusions:Our data suggest that the growth of S. aureus may be enhanced by β lactams (methicillin) and that MSSA may also display resistance to methicillin and perhaps other β lactam antibiotics. The high prevalence of MDEs suggestive of resistance to a broad spectrum of biocides and fluoroquinolones are particularly disturbing.

Antimicrobial Resistance and Molecular Characteristics of Methicillin-Resistant Staphylococcus aureus Isolates from Child Patients of High-Risk Wards in Shenzhen, China.

Methicillin-resistant Staphylococcus aureus (MRSA) strains are responsible for high rates of mortality and thus pose a substantial burden to public health worldwide. Here, we investigated the antimicrobial susceptibility and molecular characteristics of MRSA isolated from child patients at Shenzhen Children's Hospital. We characterized 140 MRSA strains through antimicrobial susceptibility testing. We further performed spa typing, multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCCmec) analysis, pvl gene analysis, and pulsed-field gel electrophoresis (PFGE). The analyzed MRSA strains were found to be sensitive to most non-β-lactam antimicrobial agents. Sequence type (ST) 59 was found to be the most common MLST lineage (54.3%). Most MRSA isolates belonged to the SCCmec IV (64.3%) and V (22.8%) types. The MRSA-ST59-SCCmec IV-t437 clone was the most predominant strain that infected 28.6% of all patients studied. Moreover, 50.7% of MRSA isolates were found to be pvl-positive. We report preliminary data on the prevalence and distribution of MRSA genotypes in Shenzhen Children's Hospital. We characterized MRSA colonization dynamics in child patients in China, and our findings can serve as the basis for the development of strategies to prevent MRSA infection and transmission.

Occurrence of methicillin-resistant Staphylococcus aureus in dairy cattle herds, related swine farms, and humans in contact with herds

In this study we investigated the circulation of methicillin-resistant Staphylococcus aureus (MRSA) in 2 dairy cattle farms (farm A and B), previously identified as MRSA-positive in bulk tank milk samples, and epidemiologically related to swine farms. Collected specimens included quarter milk samples and nasal swabs from dairy cows, pig nasal swabs collected at both the farm and slaughterhouse level, environmental dust samples, and human nasal swabs from the farms’ owners and workers. The prevalence of MRSA was estimated at the herd level by testing quarter milk samples. The prevalence of MRSA was 4.8% (3/63; 95% confidence interval=0–10.2%) and 60% (33/55; 95% confidence interval=47.05–72.95) in farm A and B, respectively. In farm A, MRSA was also isolated from humans, pigs sampled at both farm and slaughterhouse level, and from environmental samples collected at the pig facilities. The dairy cattle facilities of farm A tested negative for MRSA. In farm B, MRSA was isolated from environmental dust samples in both the cattle and pig facilities, whereas nasal swabs collected from cows and from humans tested negative. Sixty-three selected MRSA isolates obtained from different sources in farm A and B were genetically characterized by multilocus sequence typing, spa-typing, ribosomal spacer-PCR, and also tested for the presence of specific virulence genes and for their phenotypical antimicrobial susceptibility by broth microdilution method. Different clonal complex (CC) and spa-types were identified, including CC398, CC97, and CC1, CC already reported in livestock animals in Italy. The MRSA isolates from quarter milk of farm A and B mostly belonged to CC97 and CC398, respectively. Both lineages were also identified in humans in farm A. The CC97 and CC398 quarter milk isolates were also identified as genotype GTBE and GTAF by ribosomal spacer-PCR respectively, belonging to distinct clusters with specific virulence and resistance patterns. The GTBE and GTAF clusters also included swine, environmental, and human isolates from both farms. A high heterogeneity in the genetic and phenotypic profiles was observed in environmental isolates, in particular from farm B. These results demonstrate the possibility of a dynamic sharing and exchange of MRSA lineages or genotypes between different species and farm compartments in mixed-species farms. The risk of transmission between swine and related dairy cattle herds should be considered. Our findings also confirm the zoonotic potential of livestock-associated MRSA and underline the importance of applying biosecurity measures and good hygiene practices to prevent MRSA spread at the farm level and throughout the food production chain.

Living Conditions as a Driving Factor in Persistent Methicillin-resistant Staphylococcus aureus Colonization Among HIV-infected Youth

Methicillin-resistant Staphylococcus aureus (MRSA) colonization has been linked to HIV-related sexual and social behaviors. MRSA risk factors may be different for HIV-infected children, adolescents and young adults. We investigated the association of MRSA colonization, persistent colonization and genotypes with potential risk factors among HIV-infected youth. For this case-control study, patients 24 years of age or younger attending 2 HIV reference centers were recruited from February to August 2012 and followed for 1 year. Nasal swabs were collected at enrollment and every 3 months. MRSA clones were characterized by staphylococcal chromosomal cassette mec typing, spa typing and multilocus sequence typing. We compared MRSA colonization and persistent colonization with patient demographic and clinical characteristics. Among 117 participants, MRSA colonization frequency (calculated for each collection based on the number of positive cultures per patient) was 12.8% at the first collection. The average MRSA colonization frequency was 10.4%. Our results showed 11.1% were persistent carriers (subjects with more than 1 positive culture in at least 3). Crowding was the only factor associated with MRSA colonization (P = 0.018). Persistent carriers had significantly higher (4.2 times) odds of living in a crowded household (95% confidence interval-1.1-16.2). We observed high genetic diversity among MRSA isolates, with t002/ST5 and t318/ST30 being the most frequent. MRSA colonization among HIV-infected youth is more closely related to living in a low-income or slum community than to HIV-related clinical factors. High genetic MRSA isolate diversity in our population suggests frequent transmission.

Prevalence and genotype distribution of methicillin-resistant Staphylococcus aureus (MRSA) in India.

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious human pathogen that can cause a wide variety of infections. Comparative genetic analyses have led to the discovery that despite the existence of a vast number of genotypes, outbreak strains of MRSA appear to be limited to certain genotypes, some of which are further restricted to certain geographical locations. Whereas extensive literature is available in several countries, the complexity of the clonal distribution both of healthcare-associated (HA) and community-associated (CA) MRSA in India is only now beginning to be understood. Studies have revealed that MRSA in India is distributed among all of the major staphylococcal cassette chromosome mec (SCCmec) types. The majority of HA-MRSA isolates belong to SCCmec type III and sequence type (ST) 239. By contrast, CA-MRSA mostly belong to ST22 (SCCmec IV), ST772 (SCCmec V) and ST672 (SCCmec V) genotypes. Similar to the global scenario, CA-MRSA is becoming more invasive and transmissible and is increasingly becoming difficult to be differentiated from HA-MRSA. In addition, it is disturbing that some of the HA-MRSA isolates have been reported to be vancomycin-resistant. On the other hand, almost no information is available on the genotypes of livestock-associated MRSA or their potential impact on human infections in India. Concerted efforts are needed to further understand the genetic epidemiology of MRSA in India.

Determinants of Outcome in Hospitalized Patients With Methicillin-Resistant Staphylococcus aureus Bloodstream Infection: Results From National Surveillance in Canada, 2008-2012

BACKGROUND Bloodstream infection (BSI) due to methicillin-resistant Staphylococcus aureus (MRSA) is associated with considerable morbidity and mortality. OBJECTIVE To determine the incidence of MRSA BSI in Canadian hospitals and to identify variables associated with increased mortality. METHODS Prospective surveillance for MRSA BSI conducted in 53 Canadian hospitals from January 1, 2008, through December 31, 2012. Thirty-day all-cause mortality was determined, and logistic regression analysis was used to identify variables associated with mortality. RESULTS A total of 1,753 patients with MRSA BSI were identified (incidence, 0.45 per 1,000 admissions). The most common sites presumed to be the source of infection were skin/soft tissue (26.6%) and an intravascular catheter (22.0%). The most common spa types causing MRSA BSI were t002 (USA100/800; 55%) and t008 (USA300; 29%). Thirty-day all-cause mortality was 23.8%. Mortality was associated with increasing age (odds ratio, 1.03 per year [95% CI, 1.02-1.04]), the presence of pleuropulmonary infection (2.3 [1.4-3.7]), transfer to an intensive care unit (3.2 [2.1-5.0]), and failure to receive appropriate antimicrobial therapy within 24 hours of MRSA identification (3.2 [2.1-5.0]); a skin/soft-tissue source of BSI was associated with decreased mortality (0.5 [0.3-0.9]). MRSA genotype and reduced susceptibility to vancomycin were not associated with risk of death. CONCLUSIONS This study provides additional insight into the relative impact of various host and microbial factors associated with mortality in patients with MRSA BSI. The results emphasize the importance of ensuring timely receipt of appropriate antimicrobial agents to reduce the risk of an adverse outcome.