scholarly journals IDENTIFICATION AND DETECTION OF BIOFILM PRODUCING STAPHYLOCOCCUS AUREUS AND ITS ANTIBIOGRAM ACTIVITIES

Author(s):  
SAPANA SHARMA ◽  
UPASHANA BHANDARI ◽  
YOGESH OLI ◽  
GANESH BHANDARI ◽  
SUNITA BISTA ◽  
...  

Objectives: The main aim of this work is to determine the antibiogram profile of biofilm-producing Staphylococcus aureus from various clinical specimens of the patients. Methods: Various bacterial cultures of non-repeated clinical specimens from a total of 3388 patients were determined using standard microbiological and biochemical methods. Results: Out of 3388 only 604 (17.02%) displayed growth positive. A total of 65 (51.58%) S. aureus isolates were recovered, 25 (38.46%) were identified as methicillin-resistant S. aureus (MRSA) by Cefoxitin (30 μg) disk diffusion technique, of which majority were from pus/wound swab 22 (37.29%). The antibiogram of the isolates was analyzed by Kirby-Bauer disk diffusion technique analyzing Linezolid to be the most effective drug with susceptibility of 100% to both MRSA and methicillin-sensitive S. aureus, followed by vancomycin, tigecycline, and tetracycline. In vitro biofilm production by tissue culture plate (TCP) and Congo red agar method detected 52 (80%) and 25 (38.46%) as biofilm producers, respectively. TCP identified 2 (3.07%), 7 (10.76%), and 44 (67.69%) as strongly, moderately, and weakly adherent. About 30.7% of MRSA obtained were positive biofilm producers. The minimum inhibitory concentration value of Oxacillin for S. aureus by agar dilution method ranged from 0.025 μg/mL to 128 μg/mL. Conclusion: This study shows that biofilm production was more in methicillin-resistant strains and displayed a high degree of resistance to almost all groups of antibiotics.

2011 ◽  
Vol 6 (04) ◽  
pp. 317-323 ◽  
Author(s):  
Ezekiel Olugbenga Akinkunmi ◽  
Adebayo Lamikanra

Introduction: The study aimed to investigate the resistance of methicillin resistant Staphylococcus aureus (MRSA), an indicator used in hospitals but isolated from faecal samples of children in the community, to commonly used antibiotics and antiseptic agents. Methodology: S. aureus isolates were identified by phenotypic and genotypic techniques such as biochemical tests and polymerase chain reaction. Antibiotic susceptibility was investigated using the disc diffusion technique while the agar dilution method was used to determine the minimum inhibitory concentration (MIC) of antiseptics. Results: MRSA showed considerably higher resistance to other antibiotics than methicillin sensitive Staphylococcus aureus (MSSA). Twelve percent of the MSSA were susceptible to all the antibiotics studied while none of the MRSA had this property. A significant difference in susceptibility between MRSA and MSSA to the three antiseptic agents was observed as 68.8%, 75.0% and 100% of MRSA were less susceptible to benzalkonium chloride, chlorhexidine and cetrimide respectively, while 32.0%, 28.0% and 56.0% of MSSA respectively were less susceptible to these agents compared with S. aureus NCTC 6571. Overall, the MICs for the antiseptics were 2-3 times greater in the MRSA than in the MSSA (p < 0.001). Conclusion: Results show that the concentration of antiseptics used in the prevention of the transmission of infectious agents may have to be raised to cope with the possible presence of MRSA in patients coming into hospital.


2014 ◽  
Vol 52 (196) ◽  
pp. 977-981 ◽  
Author(s):  
Prakash Chandra Pahadi ◽  
Upendra Thapa Shrestha ◽  
Nabaraj Adhikari ◽  
Pradeep Kumar Shah ◽  
Ritu Amatya

Introduction: Methicillin resistant Staphylococcus aureus (MRSA), majorly associated with nosocomial and community infections worldwide, are emerging as resistant strains to many antibiotics narrowing down the efficacy of antimicrobial therapy. In order to investigate the changing resistant pattern of MRSA to empirical drugs, the study was carried out at KIST Medical College and Hospital, Nepal. It also aims to determine the minimum inhibitory concentration of vancomycin among MRSA. Methods: Altogether 3500 clinical samples including 1303 blood, 1489 urine and 708 body fluids were collected and processed. Isolated S. aureus were further screened for methicillin resistance by Kirby-Bauer disk diffusion technique using cefoxitin (30μg) disk. All MRSA were subjected to in vitro determination of MIC of vancomycin by agar dilution method as recommended by CLSI guidelines. Results: Total 287 S. aureus were isolated from the different clinical samples. Altogether 248 (86.41%) were found to be multidrug resistance (MDR) while 42 (14.63%) of the isolates were methicillin resistance with the highest prevalence in the age group of 16-30. All 42 (100%) MRSA isolates were resistant to ampicillin and penicillin followed by 41 (97.62%), 32 (76.19%), 31(73.81%), 29 (69.05%), 9 (21.43%) and seven (16.67%) to cefotaxime, gentamycin, cotrimoxazole, erythromycin, tetracycline and ciprofloxacin respectively. Although all MRSA strains were sensitive to vancomycin on disc diffusion, four isolates were intermediates in vitro determination of MIC of vancomycin. The break point for vancomycin was found to be 15mm. Conclusions: The increment in vancomycin MIC among MRSA is alarming. Strict control measures to prevent MRSA spread and a routine surveillance for VRSA must be incorporated in hospitals.  Keywords: mdr; mrsa; mic; visa; vrsa.


2006 ◽  
Vol 50 (8) ◽  
pp. 2680-2685 ◽  
Author(s):  
Olivier Denis ◽  
Ariane Deplano ◽  
Claire Nonhoff ◽  
Marie Hallin ◽  
Raf De Ryck ◽  
...  

ABSTRACT The in vitro activities of 22 antimicrobial agents, including ceftobiprole, daptomycin, and tigecycline, against 511 methicillin-resistant Staphylococcus aureus (MRSA) isolates from 112 Belgian hospitals were studied by using the CLSI agar dilution method. Isolates were characterized by pulsed-field gel electrophoresis (PFGE) analysis and by PCR detection of determinants of resistance to aminoglycosides, macrolides-lincosamides-streptogramins, and tetracyclines. A representative set of isolates with different PFGE genotypes was further characterized by multilocus sequence typing, determination of staphylococcal cassette chromosome mec (SCCmec) type, and multiplex PCR for toxic shock syndrome type 1 (TSST-1) and Panton-Valentine leukocidin genes. MRSA isolates belonged to nine epidemic MRSA clones, of which sequence type 45 (ST45)-SCCmec IV and ST8-SCCmec IV were predominant, accounting for 49 and 20% of isolates, respectively. The distribution of antimicrobial resistance and TSST-1 genes was strongly linked to clonal types. Ceftobiprole, daptomycin, and tigecycline showed high activity against all isolates of these sporadic and epidemic MRSA clones, as indicated by MIC90s of 2 mg/liter, 0.5 mg/liter, and 0.25 mg/liter, respectively. The MIC distribution of daptomycin and tigecycline was not different in isolates with decreased susceptibility to glycopeptides or tetracyclines, respectively. Ceftobiprole MICs were not correlated with oxacillin and cefoxitin MICs. These data indicate excellent activity of the newly developed agents ceftobiprole, daptomycin, and tigecycline against MRSA isolates recently recovered from hospitalized patients in Belgium, supporting their therapeutic potential for nosocomial MRSA infections.


2021 ◽  
Vol 71 (1) ◽  
pp. 150-54
Author(s):  
Sheroze Ilyas ◽  
Tehmina Munir ◽  
Rabia Sadaf ◽  
Mehreen Gilani

Objective: To compare the in-vitro efficacy by determining Minimum Inhibitory Concentration of Vancomycinusing the reference Agar Dilution to the E-Strip in Methicillin Resistant Staphylococcus aureus isolates. Study Design: Validation study. Place and Duration of Study: The department of Microbiology Army Medical College/National University ofMedical Sciences in collaboration with Pak Emirates Military Hospital Rawalpindi, from Dec 2016 to Dec 2017. Methodology: Non-duplicate 84 isolates of Methicillin resistant Staphylococcus aureus from various clinical specimens were included in the study. All these isolates were screened for susceptibility to glycopeptide by E-strips method (Bio mérieux) as well as Agar Dilution method, using vancomycin concentrations of 0.25, 0.50, 1.00, 2.00, 4.00 and 8.00µgm/ml respectively in two fold serial dilutions. Results: There was an overall agreement on 83 samples by both the methods i.e. 83 were Vancomycin SensitiveStaphylococcus aureus by both methods while one isolate with intermediate resistance to Vancomycin was onlydetected by Agar Dilution. The sensitivity of the E–strips compared to Agar Dilution was found to be 100%. Thepositive predictive value was 98.8% with a diagnostic accuracy of 98.8%. Specificity and negative predictive valuecould not be ascertained for E-strips because of the limitation of the method to detect the Vancomycin Intermediate Staphylococcus aureus isolates. Conclusion: E-strip can be a convenient alternative to the gold standard Agar Dilution but its inability to identifyVISA challenges its reliability in determining the Vancomycin resistance in MRSA isolates.


2015 ◽  
Vol 59 (10) ◽  
pp. 6262-6265 ◽  
Author(s):  
Ko-Hung Chen ◽  
Yu-Tsung Huang ◽  
Chun-Hsing Liao ◽  
Wang-Hui Sheng ◽  
Po-Ren Hsueh

ABSTRACTTedizolid is a novel, expanded-spectrum oxazolidinone with potent activity against a wide range of Gram-positive pathogens. A total of 425 isolates of Gram-positive bacteria were obtained consecutively from patients with acute bacterial skin and skin structure infections (ABSSSIs) or pneumonia. These isolates included methicillin-susceptibleStaphylococcus aureus(MSSA) (n= 100), methicillin-resistantStaphylococcus aureus(MRSA) (n= 100),Streptococcus pyogenes(n= 50),Streptococcus agalactiae(n= 50),Streptococcus anginosusgroup (n= 75),Enterococcus faecalis(n= 50), and vancomycin-resistant enterococci (VRE) (Enterococcus faecium) (n= 50). The MICs of tedizolid and linezolid were determined by the agar dilution method. Tedizolid exhibited betterin vitroactivities than linezolid against MSSA (MIC90s, 0.5 versus 2 μg/ml), MRSA (MIC90s, 0.5 versus 2 μg/ml),S. pyogenes(MIC90s, 0.5 versus 2 μg/ml),S. agalactiae(MIC90s, 0.5 versus 2 μg/ml),Streptococcus anginosusgroup (MIC90s, 0.5 versus 2 μg/ml),E. faecalis(MIC90s, 0.5 versus 2 μg/ml), and VRE (MIC90s, 0.5 versus 2 μg/ml). The tedizolid MICs againstE. faecalis(n= 3) and VRE (n= 2) intermediate to linezolid (MICs, 4 μg/ml) were 1 μg/ml and 0.5 μg/ml, respectively. The tedizolid MIC90s against S. anginosus,S. constellatus, andS. intermediuswere 0.5, 1, and 0.5 μg/ml, respectively, and the rates of susceptibility based on the U.S. FDA MIC interpretive breakpoints to the isolates were 16%, 28%, and 72%, respectively. Tedizolid exhibited 2- to 4-fold betterin vitroactivities than linezolid against a variety of Gram-positive cocci associated with ABSSSIs and pneumonia. The lower susceptibilities of tedizolid against isolates ofS. anginosusandS. constellatusthan against those ofS. intermediusin Taiwan were noted.


2019 ◽  
Vol 57 (9) ◽  
Author(s):  
Ayesha Khan ◽  
Lina M. Rivas ◽  
Maria Spencer ◽  
Rodrigo Martinez ◽  
Marusella Lam ◽  
...  

ABSTRACT Ceftaroline (CPT) is a broad-spectrum agent with potent activity against methicillin-resistant Staphylococcus aureus (MRSA). The sequence type 5 (ST5) Chilean-Cordobés clone, associated with CPT nonsusceptibility, is dominant in Chile, a region with high rates of MRSA infections. Here, we assessed the in vitro activity of CPT against a collection of MRSA isolates collected between 1999 and 2018 from nine hospitals (n = 320) and community settings (n = 41) in Santiago, Chile, and evaluated performance across testing methodologies. We found that our hospital-associated isolates exhibited higher CPT MIC distributions (MIC50 and MIC90 of 2 mg/liter) than the community isolates (MIC50 and MIC90 of 0.5 mg/liter), a finding that was consistent across time and independent of the culture source. High proportions (64%) of isolates were CPT nonsusceptible despite the absence of CPT use in Chile. Across methodologies, the Etest underestimated the MIC relative to the gold standard broth microdilution (BMD) test (MIC50 and MIC90 of 1 and 1.5 mg/liter, respectively). There was low (∼51%) categorical agreement (CA) between Etest and BMD results across CLSI and EUCAST breakpoints. The recent revision of CLSI guidelines abolished “very major error” (VME) from the previous guidelines (81%), which perform similarly to the EUCAST guidelines. The level of concordance between CLSI and EUCAST for BMD testing and Etest was >95%. Disk diffusion performed poorly relative to BMD under CLSI (CA, 55%) and EUCAST (CA, 36%) guidelines. Comparison of EUCAST to CLSI for disk diffusion (with EUCAST used as the reference) showed low agreement (CA, 25%; VME, 70%). In summary, CPT-nonsusceptible MRSA are dominant in clinical settings in Chile. Our results provide data to support the reevaluation of CPT breakpoints and to improve agreement across methodologies and agencies.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiwei Zhong ◽  
Haidan Wang ◽  
Yun Zhuang ◽  
Qun Shen

AbstractCryptotanshinone (CT) is an extract from the traditional Chinese medicine Salvia miltiorrhiza, which inhibits the growth of methicillin-resistant Staphylococcus aureus (MRSA) in vitro. This study aims to determine the antibacterial mechanisms of CT by integrating bioinformatics analysis and microbiology assay. The microarray data of GSE13203 was retrieved from the Gene Expression Omnibus (GEO) database to screen the differentially expressed genes (DEGs) of S. aureus strains that were treated with CT treatment. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to identify the potential target of CT. Data mining on the microarray dataset indicated that pyruvate kinase (PK) might be involved in the antimicrobial activities of CT. The minimum inhibition concentrations (MICs) of CT or vancomycin against the MRSA strain ATCC43300 and seven other clinical strains were determined using the broth dilution method. The effects of CT on the activity of PK were further measured. In vitro tests verified that CT inhibited the growth of an MRSA reference strain and seven other clinical strains. CT hampered the activity of the PK of ATCC43300 and five clinical MRSA strains. CT might hinder bacterial energy metabolism by inhibiting the activity of PK.


2011 ◽  
Vol 8 (3) ◽  
pp. 1120-1123 ◽  
Author(s):  
Bahram Letafat ◽  
Negar Mohammadhosseini ◽  
Ali Asadipour ◽  
Alireza Foroumadi

In the present study we report the synthesis and antibacterial activity of a new series 2-(1-methyl-4-nitro-1H-imidazol-5-ylsulfonyl)-1,3,4-thiadiazoles (6a-c). Compounds6a-cwere testedin vitroby the conventional agar dilution method against a panel of microorganisms including gram-negative and gram-positive bacteria. Compound6bwith 5-(5-nitrofuran-2-yl)-residue on 1,3,4-thiadiazole scaffold have shown promising antibacterial activities against gram-positive bacteria includingStaphylococcus aureus, Staphylococcus epidermidisandBacillus subtilis.


Author(s):  
Swati S. Kale ◽  
Ashwini Patil

Background: Staphylococcus aureus has emerged over the past several decades as a leading cause of hospital-associated and community acquired infections. Methicillin resistant S. aureus (MRSA), which are often resistant to several classes of antibiotics, is the most common cause of nosocomial infections and pose a great threat to the world. Vancomycin is regarded as the first-line drug for treatment of MRSA but resistance to this drug is being reported now a day.Methods: It was carried out for a period between January 2014 to June 2017 in the microbiology diagnostic laboratory. MRSA detection was performed by cefoxitin disk diffusion method. Screening for the vancomycin intermediate and the vancomycin resistant S. aureus (VISA and VRSA respectively) was carried out by using vancomycin screen. MIC (minimum inhibitory concentration) of vancomycin was tested by agar dilution method and E strip on all MRSA isolates.Results: A total of 287 S. aureus clinical isolates were included in the study. All MRSA were inoculated on vancomycin screen agar. Visible growth was present in 8 isolates. Five (3.73%) MRSA isolates with MIC of 4 were termed VISA (vancomycin intermediate S. aureus) by agar dilution method. Six isolates had the MIC of 4 and were termed as VISA.Conclusions: As disc diffusion method is not recommended by CLSI for S. aureus, vancomycin screen agar and MIC determination by either of the methods viz. agar dilution or E test can be used.


2015 ◽  
Vol 13 (2) ◽  
pp. 35-38
Author(s):  
Sabita Bhatta ◽  
Babli Basu ◽  
Chandrasekhar Narharrao Chaudhary ◽  
Ashok Kumar Praharaj

Introduction: Tigecycline is a novel glycylcycline  derivative of the tetracycline with activity against a wide range of  organisms including Methicillin resistant Staphylococcus aureus, Vancomycin  resistant  Enterococcus , Extended spectrum beta lactamase   producing  (Escherichia coli , Klebsiella  pneumonia)  and Acinetobacter species.  The aim of the study was to assess effectiveness of the drug against methicillin resistant Staphylococcus aureus (MRSA), vancomycin resistant enterococci (VRE), ESBL producers and carbapenem resistant Acinetobacter baumannii and to compare the efficacy of different methods of antimicrobial susceptibility testing for Tigecycline.Methods: A total of 250 clinical isolates were processed and identified by conventional methods. In all the 250 isolates, antimicrobial susceptibility was carried out by disc diffusion method , Minimum inhibitory test by agar dilution method (MIC) and in 30 isolates of A baumannii  MIC was also done by E test.Results: Out of 250 isolates, 236 isolates were sensitive to tigecycline by agar dilution method while only 159 were sensitive by disk diffusion method.Conclusion: Marked discordance was observed between the results of two different methods (DDT & Agar dilution method) for E coli, Klebsiella spp and A baumannii, where significant number of isolates were resistant to tigecycline by DDT as compared to AD method. But results of MIC by agar dilution method & E test were in concordance for A. baumannii.


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