Methicillin-susceptible Staphylococcus Aureus Isolates Research Articles

Prevalence of the cefazolin inoculum effect (CzIE) in nasal colonizing methicillin-susceptible Staphylococcus aureus in patients from intensive care units in Colombia and use of a modified rapid nitrocefin test for detection.

The cefazolin inoculum effect (CzIE) has been associated with poor clinical outcomes in patients with methicillin-susceptible Staphylococcus aureus (MSSA) infections. We aimed to investigate the point prevalence of the CzIE among nasal colonizing MSSA isolates from ICU patients in a multicenter study in Colombia (2019-2023). Patients underwent nasal swabs to assess for S. aureus colonization on admission to the ICU, and some individuals had follow-up swabs. We performed cefazolin MIC by broth microdilution using standard and high inoculum and developed a modified nitrocefin-based rapid test to detect the CzIE. Whole-genome sequencing was carried out to characterize BlaZ types and allotypes, phylogenomics, and Agr-typing. A total of 352 patients were included; 46/352 (13%) patients were colonized with S. aureus and 22% (10/46) and 78% (36/46) with MRSA and MSSA, respectively. Among 36 patients who contributed with 43 MSSA colonizing isolates, 21/36 (58%) had MSSA exhibiting the CzIE. BlaZ type A and BlaZ-2 were the predominant type and allotype in 56% and 52%, respectively. MSSA belonging to CC30 were highly associated with the CzIE, and single-nucleotide polymorphism (SNP) analyses supported possible transmission of MSSA exhibiting the CzIE among some patients of the same unit. The modified nitrocefin rapid test had 100%, 94.4%, and 97.7% sensitivity, specificity, and accuracy, respectively. We found a high point prevalence of the CzIE in MSSA colonizing the nares of critically ill patients in Colombia. A modified rapid test was highly accurate in detecting the CzIE in this patient population.

Genomic profiling of methicillin-sensitive Staphylococcus aureus (MSSA) isolates in Kuwait hospitals.

Staphylococcus aureus is an important pathogen that causes mild to invasive infections in hospitals and the community. Although methicillin-susceptible Staphylococcus aureus (MSSA) isolates continue to cause different infections, there is no data on the genetic backgrounds of the MSSA colonizing or causing infections in Kuwait hospitals. This study aimed to investigate MSSA isolated from patients admitted to Kuwait hospitals for antibiotic resistance and genetic backgrounds to understand their clonal composition. Consecutive MSSA isolates were collected from single patients during two surveillance periods in 2016 and 2021 in 13 public hospitals. The isolates were characterized using antibiogram, staphylococcal protein A (spa) typing, DNA microarray analysis, and multilocus sequence typing (MLST) using standard protocols. A total of 446 MSSA was cultured from different clinical samples in 2016 (n = 240) and 2021 (n = 206). All isolates were susceptible to vancomycin [minimum inhibitory concentration (MIC) ≤ 2 mg/L], teicoplanin (MIC ≤2 mg/L), linezolid (MIC ≤4 mg/L), ceftaroline (MIC ≤2 mg/L), rifampicin, and mupirocin but were resistant to erythromycin (21.3%), clindamycin (14.0%), gentamicin (3.8%), kanamycin (10.5%), fusidic acid (27.0%), tetracycline (6.9%), trimethoprim (23.1%), and ciprofloxacin (35.2%). Molecular typing identified 155 spa types, dominated by t127 (15.0%), t084 (5.4%), t3841 (5.4%), t267 (2.4%), t442 (2.2%), t091 (2.2%), t021 (2.2%), and t003 (2.2%); 31 clonal complexes (CCs); and 56 sequence types (STs). The majority of the isolates (n = 265; 59.4%) belonged to CC1 (20.6%), CC15 (10.9%), CC22 (5.1%), CC30 (7.6%), CC361 (10.1%), and CC398 (4.7%). The MSSA isolates belonged to diverse genetic backgrounds dominated by CC1, CC15, CC22, CC30, CC361, and CC398. The distribution of MSSA clones in 2016 and 2021 showed the stability of these clones over time. The study provides the first comprehensive data on the clonal distribution of MSSA in Kuwait hospitals.

In Vitro Activity of Ceftaroline and Comparators against Bacterial Isolates Collected Globally from Patients with Skin and Soft Tissue Infections: ATLAS Program 2019-2020.

The objective of this study was to assess the in vitro activity of ceftaroline and a panel of comparator agents against isolates causing skin and soft tissue infections (SSTIs) collected in Africa/Middle East, Asia-Pacific, Europe, and Latin America from 2019-2020. Minimum inhibitory concentrations (MIC) were determined using European Committee on Antimicrobial Susceptibility Testing criteria. All the methicillin-susceptible Staphylococcus aureus (MSSA) isolates were susceptible to ceftaroline. Across all regions, ceftaroline demonstrated potent activity against methicillin-resistant S. aureus (MRSA, susceptibility 89.5-93.7%) isolates. Susceptibility to vancomycin, daptomycin, linezolid, teicoplanin, trimethoprim sulfamethoxazole, and tigecycline was ≥94.1% in MSSA and MRSA isolates. Against β-hemolytic streptococci isolates, ceftaroline demonstrated very potent activity (MIC90 0.008-0.03 mg/L) across all regions. All β-hemolytic streptococci isolates were susceptible to linezolid, penicillin, and vancomycin (MIC90 0.06-2 mg/L). Among the extended-spectrum β-lactamases (ESBL)-negative Enterobacterales tested (E. coli, K. pneumoniae, and K. oxytoca), susceptibility to ceftaroline was high (88.2-98.6%) in all regions. All ESBL-negative Enterobacterales were susceptible to aztreonam. Potent activity was observed for amikacin, cefepime, and meropenem (94.1-100%) against these isolates. Overall, ceftaroline showed potent in vitro activity against isolates of pathogens causing SSTIs. Continuous surveillance of global and regional susceptibility patterns is needed to guide appropriate treatment options against these pathogens.

Detecting the performance of methicillin-resistant Staphylococcus aureus by a molecular diagnostic assay in positive blood culture: Influence of coexistence of mecA-positive bacteria and diversity in orfX-SCCmec junction region in methicillin-susceptible S. aureus

BackgroundIn blood cultures that test positive for staphylococcal bacteria, rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) or methicillin-susceptible Staphylococcus aureus (MSSA) by molecular assay is useful for appropriate antimicrobial treatment of bloodstream infections. Although the Xpert MRSA/SA BC assay is widely available in clinical settings in Japan, its efficacy has not yet evaluated thoroughly. MethodsWe retrospectively studied 100 blood culture cases positive for S. aureus at Sapporo Medical University Hospital between March 2019 and May 2022. Cycle threshold (CT) values for target genes from the Xpert MRSA/SA BC assay were compared to phenotypic results. Genotyping and genetic analysis of the orfX-SCCmec junction region was performed for selected isolates. ResultsWe analyzed 25 and 75 isolates assigned to MRSA and MSSA, respectively, using the Xpert MRSA/SA BC assay. Of these, 99 isolates from agar cultures showed compatible susceptibility to oxacillin. One genetically misidentified case of MRSA was found to be caused by the mixed growth of MSSA and methicillin-resistant S. hominis on agar culture. Of the 73 MSSA with pure growth on agar culture, 45 (61.6%) were found to be orfX-SCCmec-positive, spa-positive, and mecA-negative in this assay. These MSSA belong to diverse spa and coa types. ConclusionThe Xpert MRSA/SA BC assay accurately identified MRSA and MSSA in positive blood cultures. However, over half of the MSSA isolates showed positive results for orfX-SCCmec, presumably due to genetic diversity in the orfX-associated region of MSSA. Therefore, the coexistence of MSSA and mecA-harboring coagulase-negative staphylococci may cause confusion about identification of MRSA.

The phenotypes and genotypes associated with biofilm formation among methicillin-susceptible Staphylococcus aureus (MSSA) isolates collected from a tertiary hospital in Terengganu, Malaysia.

Methicillin-susceptible Staphylococcus aureus (MSSA) is an important nosocomial pathogen worldwide. This study aims to investigate the in vitro biofilm-forming ability of clinical MSSA isolated from various sources in the main public tertiary referral hospital in Terengganu, Malaysia and to detect the presence of biofilm-associated and regulatory genes among these isolates. A total of 104 MSSA isolates [pus (n = 75), blood (n = 24), respiratory secretions (n = 2), eye (n = 2), and urine (n = 1)] were investigated for slime production and biofilm formation using Congo red agar and crystal violet microtitre plate, respectively. Fifteen MSSA isolates with varying degrees of biofilm formation were selected for validation via a real-time cell analyser. All isolates were screened for microbial surface components recognising adhesive matrix molecules (MSCRAMM) and accessory gene regulator (agr) using polymerase chain reaction assay. A total of 76.0% (79/104) isolates produced slime layer, while all isolates developed biofilm as follows: 52.8% (55/104) strong biofilm producers, 40.4% (42/104) intermediate biofilm producers, and 6.7% (7/104) weak biofilm producers. A total of 98.1% (102/104) isolates carried at least one of the screened MSCRAMM gene(s) with the eno gene detected at the highest rate (87.5%, 91/104), while the sasG gene was significantly associated with strong biofilm production (p = 0.015). Three agr groups, 1, 2, and 3, were detected among the MSSA isolates with a predominance of agr-3 (32.7%, 34/104). In conclusion, biofilm formation varied greatly among clinical MSSA isolates, and the presence of sasG gene and agr-1 may play important role in initiating MSSA infections via biofilm formation.

PREVALENCE OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS AND OTHER STAPHYLOCOCCAL NASAL CARRIAGES AMONG HEALTHCARE WORKERS, PHRAMONGKUTKLAO HOSPITAL

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a group of S. aureus strains containing the SCCmec gene causing beta-lactam antibiotic resistance. MRSA is common in healthcare settings and can cause serious problems.
 Objective: The study aimed to investigate the prevalence of MRSA nasal colonization among privates of the Medical Private Company, Phramongkutklao Hospital, including antibiotic susceptibility pattern of S. aureus isolates and risk factors of S. aureus nasal carriage.
 Methods: Nasal swabs were obtained from the anterior nares of 170 privates. Staphylococcal isolates were identified using a catalase test, tube coagulase test and matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF MS). MRSA detection was screened using cefoxitin disk diffusion and confirmed using the mecA gene detection and SCCmec typing. Antibiotic susceptibility patterns of S. aureus were examined using the disk diffusion method. A questionnaire was collected from the subjects to determine risk factors for S. aureus nasal carriage.
 Results: Of 170 subjects, 157 (92.35%) revealed staphylococcal positive, yielding 161 staphylococcal isolates. The prevalence of MRSA, methicillin-resistant Staphylococcus epidermidis (MRSE), and methicillin-susceptible Staphylococcus aureus (MSSA) nasal carriage was 0.59, 1.18 and 8.82%, respectively. The MRSA isolate carried mecA revealing SCCmec type II. The MSSA isolates indicated low resistance to tetracycline (13.3%), whereas the MRSA isolate resisted ciprofloxacin, clindamycin, erythromycin, gentamicin, oxacillin and tetracycline. Using multiple logistic regression analysis, a significant risk factor for S. aureus nasal carriage was utensil sharing (adjusted odds ratio=4.41; 95% CI=1.33-14.61).
 Conclusion: Healthcare-associated MRSA existed among privates of the Medical Private Company. An associated risk factor for acquiring S. aureus was utensil sharing which could be used to help improve prevention and control management among privates.

Methicillin resistance Staphylococcus aureus nasal carriage and its associated factors among HIV patients attending art clinic at Dessie comprehensive specialized hospital, Dessie, North East Ethiopia.

Globally the incidence of nosocomial infections and colonization due to methicillin resistant Staphylococcus aureus (MRSA) has become greater concern. The objective of the study was to determine the prevalence and associated factors of nasal carriage of MRSA with its antimicrobial susceptibility patter among HIV patients attending ART clinic. cross-sectional study was conducted from January 01 to May 30, 2020 at Dessie comprehensive specialized hospital, north east Ethiopia. A total of 206 HIV patients were recruited by applying systematic random sampling technique. Nasal specimen was collected from both anterior nares, and inoculated directly on mannitol salt agar, MacConkey, 5% blood agar. Screening of MRSA and methicillin susceptible Staphylococcus aureus (MSSA) strain was done by using cefoxitin antibiotic disc following modified Kirby-Bauer disc diffusion technique. Bivariable and multivariable logistic regression analyses were performed to assess the associated factors with S. aureus and MRSA. study participants were in the age range between 12 and 72 years and their mean (±SD) age was 41.52 (±11.2). The rate of S. aureus and MRSA colonization was 127/206 (61.7%) and 58/206 (28.2%), respectively. Having job close contact with human [AOR = 4.41; 95% CI = 1.5-13.02; p = 0.007], picking the nose [AOR = 4.38; 95% CI = 1.34-14.29; p = 0.014] and ART failure [AOR = 7.41; 95% CI = 2.08-26.41; p = 0.002] had statistically significant association with MRSA colonization. MRSA showed resistance for tetracycline (53.4%), erythromycin (84.5%), and trimethoprim-sulfamethoxazole (86.2%). Multi-Drug Resistance (MDR) was detected among 96.5% of MRSA and 20.3% of MSSA isolates. the rate of S. aureus and MRSA nasal colonization was high and it has associated with different factors. Understanding and managing MRSA among HIV patients is mandatory and stakeholders should find out the way how to decolonize the bacteria from nasal area.

Pharmacodynamics of Flucloxacillin in a Neutropenic Murine Thigh Infection Model: A Piece of the Puzzle towards Evidence-Based Dosing.

For decades, flucloxacillin has been used to treat methicillin-susceptible Staphylococcus aureus (MSSA). Little is still known about its pharmacodynamics (PD). The present study aimed to determine the pharmacokinetic (PK)/PD index and the PD-index value minimally required for efficacy. MICs of 305 MSSA isolates were measured to determine the wild-type distribution. The PD of 8 S. aureus, 1 S. pyogenes, and 1 S. agalactiae isolates were evaluated in a neutropenic murine thigh infection model. Two S. aureus isolates were used in a dose-fractionation study and a dose–response analysis was performed additionally in the in vivo model. Data were analyzed with a population PK and sigmoid maximum effect model. The end of the wild-type distribution was 1 mg/L. The percentage of time the unbound concentration was above MIC (%fT > MIC) was best correlated with efficacy. For S. aureus, median %fT > 0.25 × MIC required for 1-log reduction was 15%. The value for S. pyogenes was 10%fT > MIC and for S. agalactiae 22%fT > 0.25xMIC for a 1-log reduction. The effect of flucloxacillin reached a 2-log reduction of S. aureus at 20%fT > 0.25xMIC and also for S. pyogenes and S. agalactiae, a reduction was reached. These data may serve to optimize dosing regimens currently used in humans.

Cefadroxil Comparable to Cephalexin: Minimum Inhibitory Concentrations among Methicillin-Susceptible Staphylococcus aureus Isolates from Pediatric Musculoskeletal Infections.

ABSTRACTCephalexin and cefadroxil are oral first-generation cephalosporins used to treat methicillin-susceptible Staphylococcus aureus (MSSA) infections. Despite its shorter half-life, cephalexin is more frequently prescribed, although cefadroxil is an appealing alternative, given its slower clearance and possibility for less frequent dosing. We report comparative MIC distributions for cefadroxil and cephalexin, as well as for oxacillin, cephalothin, ceftaroline, and cefazolin, for 48 unique clinical MSSA isolates from pediatric patients with musculoskeletal infections. Both cefadroxil and cephalexin had MIC50 values of 2 μg/mL and MIC90 values of 4 μg/mL. MIC50s for oxacillin, cephalothin, and ceftaroline were ≤0.25 μg/mL, and cefazolin’s MIC50 was 0.5 μg/mL. While cefadroxil and cephalexin MICs are higher than those for other active agents, the distributions of MICs for cefadroxil and cephalexin are statistically equivalent, suggesting similar in vitro MSSA activities. Cefadroxil should be further considered an alternative agent to cephalexin, although additional work is needed to identify the optimal dose and frequency of these antibiotics for the treatment of serious MSSA infections.IMPORTANCE Cephalexin and cefadroxil are oral antibiotics that are used to treat serious infections due to the bacteria MSSA. While cephalexin is used more commonly, cefadroxil is excreted from the body more slowly; this generally allows patients to take cefadroxil less frequently than cephalexin. In this study, we compared the abilities of cefadroxil, cephalexin, and several other representative intravenous antibiotics to inhibit the growth of MSSA in the laboratory. Bacterial samples were obtained from children with bone, joint, and/or muscle infections caused by MSSA. We found that cefadroxil and cephalexin inhibited the growth of MSSA at similar concentrations, suggesting similar antibacterial potencies. The selected intravenous antistaphylococcal antibiotics generally inhibited bacterial growth with lower antibiotic concentrations. Based on these results, cefadroxil should be further considered an alternative oral antibiotic to cephalexin, although future research is needed to identify the optimal dose and frequency of these antibiotics for serious infections.

Comparison of Molecular Characteristics Between Methicillin-Resistant and -Susceptible Staphylococcus aureus Clinical Isolates by Whole-Genome Sequencing.

IntroductionThe transmission of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA) are great public health concern worldwide. To better understand S. aureus evolution and dissemination, we compared the molecular features of MSSA and MRSA isolates.MethodsIn this study, 74 MSSA and 102 MRSA non-duplicate isolates were recovered from clinical samples between 2016 and 2020. Molecular epidemiology, antimicrobial resistance determinants, and virulence gene profiles were carried out by whole-genome sequencing (WGS).ResultsTwenty distinct sequence types were identified in MRSA isolates, with the most common being ST59, ST630, and ST338. The major genotypes of MSSA were ST188 and ST7. The toxin genes clfA, sek, and seq were significantly associated with MRSA, while splA/B, clfB, map, sdrC/D, and sem-sen-seo-seu were detected more frequently in MSSA isolates than MRSA (P < 0.05). The tst positive isolates were more commonly identified in CC1 and CC72, whereas lukE/D was mainly found in the CC7, CC15, CC88, and completely absent in CC59 clones.ConclusionOur results compared the genetic diversity between MRSA and MSSA strains, suggesting efforts to fight infections caused by MSSA need to be intensified due to MSSA isolates carrying wide range of virulence factors. Comparative epidemiological studies of large populations of MSSA and MRSA will be necessary in the future to understand how MSSA and MRSA populations may co-evolve and interact in the future.

APTC-C-SA01: A Novel Bacteriophage Cocktail Targeting Staphylococcus aureus and MRSA Biofilms.

The high infection and mortality rate of methicillin-resistant Staphylococcus aureus (MRSA) necessitates the urgent development of new treatment strategies. Bacteriophages (phages) have several advantages compared to antibiotics for the treatment of multi-drug-resistant bacterial infections, and thus provide a promising alternative to antibiotics. Here, S. aureus phages were isolated from patients and environmental sources. Phages were characterized for stability, morphology and genomic sequence and their bactericidal activity against the biofilm form of methicillin-susceptible Staphylococcus aureus (MSSA) and MRSA was investigated. Four S. aureus phages were isolated and tested against 51 MSSA and MRSA clinical isolates and reference strains. The phages had a broad host range of 82–94% individually and of >98% when combined and could significantly reduce the viability of S. aureus biofilms. The phages had a latent period of ≤20 min and burst size of >11 plaque forming units (PFU)/infected cell. Transmission electron microscopy (TEM) identified phages belonging to the family of Myoviridae. Genomic sequencing indicated the lytic nature of all four phages, with no identified resistance or virulence genes. The 4 phages showed a high complementarity with 49/51 strains (96%) sensitive to at least 2/4 phages tested. Furthermore, the frequency of bacteriophage insensitive mutant (BIM) generation was lower when the phages were combined into the phage cocktail APTC-C-SA01 than for bacteria exposed to each of the phages alone. In conclusion, APTC-C-SA01, containing four lytic S. aureus phages has the potential for further development as a treatment against MSSA and MRSA infections.

Molecular Epidemiology, Virulence Factors, Antibiotic Resistance and Risk Factors for Nasal Carriage of Staphylococcus aureus in A Teenage Student Population: High Prevalence of Oxacillin Susceptible MRSA Isolates

Background: Asymptomatic carriage of Staphylococcus aureus can lead to endogenous infections and cross-transmission to other individuals. Objectives: The prevalence, molecular epidemiology, antibiotic resistance, and risk factors for nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) were studied in school children in Ardabil, Iran. Methods: Totally, 510 nasal samples were collected during 2017. Isolates were identified and subjected to antimicrobial susceptibility testing, identification of oxacillin resistance, and molecular typing. Results: Totally, 13.5% of volunteers were positive for methicillin-susceptible Staphylococcus aureus (MSSA) and 17.5% colonized with mecA positive S. aureus strains, including 6.07% oxacillin-resistant MRSA (OR-MRSA) and 11.56% oxacillin-susceptible MRSA (OS-MRSA). Excluding β-lactam antibiotics, high resistance rate was observed for erythromycin (71%), tetracycline (25.8%), clindamycin (35%) in our isolates. Surprisingly, 11% of the isolates [OR-MRSA (25.8%), OS-MRSA (10.1%), and MSSA (5.7%) isolates] were resistant to mupirocin. Moreover, 18 (58%), 29 (49%), and 29 (42%) of OR-MRSA, OS-MRSA, and MSSA isolates were multidrug-resistant (MDR), respectively. Overall, 97.48% of isolates carried ≥ 3 toxin encoding genes. The pvl gene was found in 46 (29%) isolates. In comparison, 25.50% of MRSA (9.60% OR-MRSA and 34% OS-MRSA) and 33% of MSSA isolates carried pvl gene. SCCmec type IV had the highest rate among OR-MRSA (87%) and OS-MRSA (74.5%) isolates, which indicates CA-MRSA phenotype. Eleven and 21 spa types were identified in OR-MRSA, and OS-MRSA isolates, respectively. The most common spa types were t11332 (14.3%) and t012 (11.4%) in OS-MRSA isolates. ERIC-PCR revealed high genetic diversity among isolates. The number of students in classroom and incomplete antibiotic course were associated with OS-MRSA nasal carriage. Conclusions: This study showed a high proportion of MDR CA-MRSA nasal carriage among Iranian healthy school children community.

In Vitro Selection of High-Level Beta-Lactam Resistance in Methicillin-Susceptible Staphylococcus aureus.

Selective pressure of beta-lactams is thought to be responsible for mutation selection in methicillin-susceptible Staphylococcus aureus (MSSA). We used next-generation sequencing to compare the genomes of beta-lactamase-positive (SA0707) and -negative (SA0937) MSSA isolates with their derivatives obtained after selection with oxacillin, ceftaroline, or meropenem. Selection with oxacillin and ceftaroline caused a rapid and significant (6–8 times) increase in the minimum inhibitory concentration (MICs) of oxacillin, penicillin, amoxicillin/clavulanate, and ceftaroline against the derivatives of both isolates, associated with growth impairment. Selection with meropenem caused a limited increase in the MICs of all beta-lactams against both isolates. During the initial stages of selection (after 5–15 passages), mutations were detected only in some reads, which indicated the heterogeneity of the population; however, during the later stages, either the population reversed to the wild type or fixation of the mutation was observed in the entire population. Selection with different beta-lactams caused diverse mutational events, but common mutations were detected in gdpP, all penicillin-binding proteins, cell wall regulators (vraST, graR), and deletions in the promoter region of pbp4. Therefore, the disk diffusion test with cefoxitin does not reveal resistance associated with these mechanisms in some cases, which can lead to the failure of beta-lactam therapy.

Antimicrobial resistance and genotyping of Staphylococcus aureus obtained from food animals in Sichuan Province, China

BackgroundStaphylococcus aureus (S. aureus), especially methicillin-resistant Staphylococcus aureus (MRSA), is considered a common zoonotic pathogen, causing severe infections. The objective of this study was to investigate the antimicrobial susceptibility, resistance genes and molecular epidemiology among MRSA and methicillin-susceptible Staphylococcus aureus (MSSA) isolated from food animals in Sichuan Province, China.MethodsThis study was conducted on 236 S. aureus isolates. All isolates were subjected to antimicrobial susceptibility testing by using a standard microbroth dilution method. The Polymerase Chain Reaction (PCR) was performed to identify genes encoding the β-lactams resistance (blaZ, mecA), macrolides (ermA, ermB, ermC) and aminoglycosides (aacA-aphD). The molecular structures and genomic relatedness of MRSA isolates were determined by staphylococcal chromosome cassette mec (SCCmec) typing and pulsed-field gel electrophoresis (PFGE), respectively.ResultsAmong 236 isolates, 24 (10.17 %) were recognized as MRSA. MRSA isolates showed different resistance rates to 11 antimicrobials ranging from 33.33 to 100 %, while for MSSA isolates the rates varied from 8.02 to 91.51 %. Multi-drug resistance phenotype was found in all MRSA isolates. The ermC gene encoding macrolides-lincosamides-streptogramin B was the most prevalent gene detected in 87.29 % of the S. aureus isolates, followed by ermB (83.05 %), blaZ (63.98 %), aacA-aphD (44.07 %), ermA (11.44 %) and mecA (11.02 %) genes. The prevalence of resistance genes in MRSA isolates was significantly higher than that of MSSA. Regarding the molecular morphology, SCCmec III (12/24, 50 %) was the most common SCCmec type. Furthermore, the PFGE typing showed that 24 MRSA were divided into 15 cluster groups (A to O), the major pulsotype J encompassed 25 % of MRSA isolates.ConclusionsThe S. aureus isolates from food animals in Sichuan province of China have severe antimicrobials resistance with various resistance genes, especially MRSA isolates. Additionally, the genetic pool of MRSA isolates is diverse and complex, and further investigation is necessary.

Prevalence of PVL and LucED Genes in Methicillin-Resistant and Susceptible Isolates of Staphylococcus aureus Isolated from Burn Patients in Kermanshah, Iran

Background: The main pathogen causing various nosocomial infections, especially in burn patients, is Staphylococcus aureus. An important virulence factor of this bacterium is leukocidin, which binds to white blood cells and leads to apoptosis and necrosis. Objectives: In the present research, we aimed at investigating the prevalence of the panton-valentine leukocidin (PVL) and LucED genes in methicillin-susceptible and resistant isolates of S. aureus isolated from burn patients in Kermanshah, Iran. Methods: Overall, 73 isolates of S. aureus were collected from burn wounds and identified by specific biochemical tests. After determining the susceptibility patterns of the isolates by the disc diffusion method, the frequencies of the PVL and LucED genes were assessed using specific primers and polymerase chain reactions. Results: The rates of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA) isolates were 58.9% (43 isolates) and 41.1% (30 isolates), respectively. In MRSA isolates, the highest antibiotic resistance was to penicillin (100%) and gentamicin (81.4%). Moreover, the frequencies of the LucED and PVL genes were 76.7% (56 isolates) and 27.4% (20 isolates), respectively, and the highest frequency of the LucED and PVL genes was observed in MRSA (81.4%) and MSSA (40%) isolates, respectively. Conclusions: Increased multidrug resistance pattern among S. aureus isolates with leukocidin production has led to treatment failure and severe infections. The development of infections can be prevented by the identification of isolates carrying the leukocidin gene, and early detection of MRSA isolates is feasible by using the PVL gene.

A Test for the Rapid Detection of the Cefazolin Inoculum Effect in Methicillin-Susceptible Staphylococcus aureus.

The cefazolin inoculum effect (CzIE) has been associated with therapeutic failures and mortality in invasive methicillin-susceptible Staphylococcus aureus (MSSA) infections. A diagnostic test to detect the CzIE is not currently available. We developed a rapid (∼3 h) CzIE colorimetric test to detect staphylococcal-β-lactamase (BlaZ) activity in supernatants after ampicillin induction. The test was validated using 689 bloodstream MSSA isolates recovered from Latin America and the United States. The cefazolin MIC determination at a high inoculum (107 CFU/ml) was used as a reference standard (cutoff ≥16 μg/ml). All isolates underwent genome sequencing. A total of 257 (37.3%) of MSSA isolates exhibited the CzIE by the reference standard method. The overall sensitivity and specificity of the colorimetric test was 82.5% and 88.9%, respectively. Sensitivity in MSSA isolates harboring type A BlaZ (the most efficient enzyme against cefazolin) was 92.7% with a specificity of 87.8%. The performance of the test was lower against type B and C enzymes (sensitivities of 53.3% and 72.3%, respectively). When the reference value was set to ≥32 μg/ml, the sensitivity for isolates carrying type A enzymes was 98.2%. Specificity was 100% for MSSA lacking blaZ The overall negative predictive value ranged from 81.4% to 95.6% in Latin American countries using published prevalence rates of the CzIE. MSSA isolates from the United States were genetically diverse, with no distinguishing genomic differences from Latin American MSSA, distributed among 18 sequence types. A novel test can readily identify most MSSA isolates exhibiting the CzIE, particularly those carrying type A BlaZ. In contrast to the MIC determination using high inoculum, the rapid test is inexpensive, feasible, and easy to perform. After minor validation steps, it could be incorporated into the routine clinical laboratory workflow.

S. pseudintermedius and S. aureus lineages with transmission ability circulate as causative agents of infections in pets for years

BackgroundStaphylococcus pseudintermedius (SP) and Staphylococcus aureus (SA) are common colonizers of companion animals, but they are also considered opportunistic pathogens, causing diseases of diverse severity. This study focused on the identification and characterization of 33 coagulase-positive staphylococci isolated from diseased pets (28 dogs and five cats) during 2009–2011 in a veterinary hospital in Spain in order to stablish the circulating lineages and their antimicrobial resistance profile.ResultsTwenty-eight isolates were identified as SP and five as SA. Nine methicillin-resistant (MR) isolates (27%) carrying the mecA gene were detected (eight MRSP and one MRSA). The 55% of SP and SA isolates were multidrug-resistant (MDR). MRSP strains were typed as ST71-agrIII-SCCmecII/III-(PFGE) A (n=5), ST68-agrIV-SCCmecV-B1/B2 (n=2), and ST258-agrII-SCCmecIV-C (n=1). SP isolates showed resistance to the following antimicrobials [percentage of resistant isolates/resistance genes]: penicillin [82/blaZ], oxacillin [29/mecA] erythromycin/clindamycin [43/erm(B)], aminoglycosides [18–46/aacA-aphD, aphA3, aadE], tetracycline [71/tet(M), tet(K)], ciprofloxacin [29], chloramphenicol [29/catpC221], and trimethoprim-sulfamethoxazole [50/dfrG, dfrK]. The dfrK gene was revealed as part of the radC-integrated Tn559 in two SP isolates. Virulence genes detected among SP isolates were as follow [percentage of isolates]: siet [100], se-int [100], lukS/F-I [100], seccanine [7], and expB [7]. The single MRSA-mecA detected was typed as t011-ST398/CC398-agrI-SCCmecV and was MDR. The methicillin-susceptible SA isolates were typed as t045-ST5/CC5 (n=2), t10576-ST1660 (n=1), and t005-ST22/CC22 (n=1); the t005-ST22 feline isolate was PVL-positive and the two t045-ST45 isolates were ascribed to Immune Evasion Cluster (IEC) type F. Moreover, the t10576-ST1660 isolate, of potential equine origin, harbored the lukPQ and scneq genes. According to animal clinical history and data records, several strains seem to have been acquired from different sources of the hospital environment, while some SA strains appeared to have a human origin.ConclusionsThe frequent detection of MR and MDR isolates among clinical SP and SA strains with noticeable virulence traits is of veterinary concern, implying limited treatment options available. This is the first description of MRSA-ST398 and MRSP-ST68 in pets in Spain, as well the first report of the dfrK-carrying Tn559 in SP. This evidences that current transmissible lineages with mobilizable resistomes have been circulating as causative agents of infections among pets for years.

Effect of the vancomycin minimum inhibitory concentration on clinical outcomes in patients with methicillin-susceptible Staphylococcus aureus bacteraemia: a systematic review and meta-analysis

ObjectiveThe use of the vancomycin minimum inhibitory concentration (MIC) as a prognostic predictor in patients with methicillin-susceptible Staphylococcus aureus (MSSA) has been debated in the last decade. We performed a...

251. High prevalence of the cefazolin inoculum effect in methicillin-susceptible Staphylococcus aureus causing bloodstream infections in a hospital network in Houston, TX

BackgroundAnti-staphylococcal β-lactams, such as anti-staphylococcal penicillins (AsPen) or cefazolin are the drugs of choice for methicillin-susceptible Staphylococcus aureus (MSSA) bloodstream infections. However, cefazolin has seen increasing use due to its better tolerance, lower cost, ease of administration and possibly better outcomes when compared to AsPen. Nevertheless, its efficacy may be compromised by the cefazolin inoculum effect (CzIE), defined as an increase in the minimum inhibitory concentration (MIC) of cefazolin to ≥16mg/L when a high inoculum (5x107 CFU/ml) is present. Previous studies have suggested that the prevalence of the CzIE varies geographically, with high prevalence in some Latin American countries. Prospective data evaluating the presence of the CzIE in deep-seated MSSA infections across the United States are lacking.MethodsWe performed a prospective observational study of MSSA bacteremia in a network of 13 hospitals in Houston, TX. Patients ≥ 18 years old, with a positive blood culture with MSSA, with at least one follow-up blood culture confirming clearance of the bacteremia, who received cefazolin or nafcillin as definitive therapy (72 hours or longer after culture results known) and whose original isolate was available for evaluation of the CzIE, were included. Patients with polymicrobial BSI, or those who received another antibiotic with activity against MSSA in the definitive therapy period were excluded. Cefazolin MICs were determined by broth microdilution at standard and high inoculum.ResultsWe report the results of 50 patients enrolled from February 15, 2020-April 30, 2020. The baseline characteristics of each group are outlined in Table 1. A total of 37/50 (74%) received cefazolin as definitive therapy, and complicated bacteremia was seen in 27/50 (54%). A total of 16/50 (32%) of the MSSA isolates exhibited the CzIE. Two patients in our cohort died: both of whose isolates exhibited the CzIE and received cefazolin as definitive therapy. ConclusionWe report a high prevalence of the CzIE in MSSA BSIs in a major US urban hospital network. Further evaluation of the clinical implications of the CzIE is urgently needed.Disclosures William R. Miller, MD, Entasis Therapeutics (Scientific Research Study Investigator)Merck (Grant/Research Support)Shionogi (Advisor or Review Panel member) Cesar A. Arias, MD, MSc, PhD, FIDSA, Entasis Therapeutics (Scientific Research Study Investigator) MeMed (Scientific Research Study Investigator) Merck (Grant/Research Support)

Molecular epidemiology of methicillin-susceptible Staphylococcus aureus in infants in a neonatal intensive care unit.

To investigate the molecular epidemiology of methicillin-susceptible Staphylococcus aureus (MSSA) in infants in a neonatal intensive care unit (NICU) using whole-genome sequencing. Investigation of MSSA epidemiology in a NICU. Single-center, level IV NICU. Universal S. aureus screening was done using a single swab obtained from the anterior nares, axilla, and groin area of infants in the NICU on a weekly basis. Core genome multilocus sequence type (cgMLST) analysis was performed on MSSA isolates detected over 1 year (2018-2019). In total, 68 MSSA-colonized infants were identified, and cgMLSTs of 67 MSSA isolates were analyzed. Overall, we identified 11 cgMLST isolate groups comprising 39 isolates (58%), with group sizes ranging from 2 to 10 isolates, and 28 isolates (42%) were unrelated to each other or any of the isolate groups. Cases of infants colonized by MSSA were scattered throughout the 1-year study period, and isolates belonging to the same cgMLST group were typically detected contemporaneously, over a few weeks or a few months. Overall, 13 infants (19.7%) developed MSSA infections: bacteremia (n = 3), wound infection (n = 5), conjunctivitis (n = 4), and cellulitis (n = 1). We detected no association between these clinically manifest infections and specific cgMLST groups. Although MSSA isolates in infants in a NICU showed high diversity, most were related to other isolates, albeit within small groups. cgMLST facilitates an understanding of the complex transmission dynamics of MSSA in NICUs, and these data can be used to inform better control strategies.