scholarly journals Biofilm Formation and Methicillin Resistance of Staphylococcus aureus Isolated from Clinical Samples

10.3823/841 ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Abdelraouf A. Elmanama ◽  
Islam Majdi Al-Aydi ◽  
Mariam R. Al-Reefi

Background: Staphylococcus aureus including methicillin resistant S. aureus (MRSA) is one of the most effective biofilm-forming organisms, biofilm contribute in protecting the microorganism from host defenses and prevent the effective penetration of antimicrobial agents. Biofilm formation is considered as an important contributing factor for the initiation and establishment of chronic infection by S. aureus and known as a major obstacle in the treatment of S. aureus infections is their ability to develop resistance to antimicrobials. Aims : To screen clinical Staphylococcus aureus including MRSA isolates for their biofilm forming abilities and their association with antimicrobial resistance. Methods: A total of 196 clinical isolates of S. aureus were obtained from different sample sources using standard microbiological techniques from three major hospitals in Gaza strip. Biofilm formation of these isolates was determined by tissue culture plate (TCP) method and tube adherence method (TM). Antimicrobial susceptibility test was performed using the modified Kirby–Bauer disk diffusion method as per Clinical and Laboratory Standards Institute guidelines. MRSA was detected using the cefoxitin disk test. Results: Biofilm formation was observed in 174 (88.8 %) and in 145 (74.0%) isolates of S. aureus via TCP method and TM, respectively. The highest resistance percent was for penicillin (92.9%), followed by cefoxitin (80.6%) and oxacillin (67.9%), while the lowest resistance percent was for linezolid and ceftaroline (1%). Among the 196 isolates 71.4% (N=140) were classified as MDR with a MAR index (≥ 0.2). A total of 158 isolates (80.6%) were identified as MRSA distributed as 90.4%, 79.4% and 70.9% from Al-Shifa, Al-Nasser pediatrics and Al-Aqsa hospitals respectively. Large proportions (82.1%) of biofilm producers were identified as MRSA. Biofilm-producing MRSA exhibited a higher percent (90.5%) when compared with the biofilm non-producer MRSA (9.5%). Importantly, 89.2% of biofilm-producing S. aureus were multidrug resistant. Conclusions: S. aureus isolates possessed high biofilm-forming ability. Biofilm-producing strains have very high tendency to exhibit antimicrobial resistance, multidrug resistance and methicillin resistance. Regular surveillance of biofilm formation by S. aureus and their antimicrobial resistance profile may lead more success in treating S. aureus infections.   Keywords: Biofilm, MDR, MRSA, TCP, TM, Gaza strip, Palestine.

2012 ◽  
Vol 45 (2) ◽  
pp. 189-193 ◽  
Author(s):  
Karinne Spirandelli Carvalho Naves ◽  
Natália Vaz da Trindade ◽  
Paulo Pinto Gontijo Filho

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is spread out in hospitals across different regions of the world and is regarded as the major agent of nosocomial infections, causing infections such as skin and soft tissue pneumonia and sepsis. The aim of this study was to identify risk factors for methicillin-resistance in Staphylococcus aureus bloodstream infection (BSI) and the predictive factors for death. METHODS: A retrospective cohort of fifty-one patients presenting bacteraemia due to S. aureus between September 2006 and September 2008 was analysed. Staphylococcu aureus samples were obtained from blood cultures performed by clinical hospital microbiology laboratory from the Uberlândia Federal University. Methicillinresistance was determined by growth on oxacillin screen agar and antimicrobial susceptibility by means of the disk diffusion method. RESULTS: We found similar numbers of MRSA (56.8%) and methicillin-susceptible Staphylococcus aureus (MSSA) (43.2%) infections, and the overall hospital mortality ratio was 47%, predominantly in MRSA group (70.8% vs. 29.2%) (p=0.05). Age (p=0.02) was significantly higher in MRSA patients as also was the use of central venous catheter (p=0.02). The use of two or more antimicrobial agents (p=0.03) and the length of hospital stay prior to bacteraemia superior to seven days (p=0.006) were associated with mortality. High odds ratio value was observed in cardiopathy as comorbidity. CONCLUSIONS: Despite several risk factors associated with MRSA and MSSA infection, the use of two or more antimicrobial agents was the unique independent variable associated with mortality.


2021 ◽  
Vol 9 (11) ◽  
pp. 2274
Author(s):  
Aya Abd El rahman ◽  
Yasmine El kholy ◽  
Rania Y. Shash

Methicillin resistance in Staphylococcus aureus has become prevalent globally. Moreover, biofilm-formation makes it more difficult to eradicate bacteria by antibiotics. The mazEF toxin-antitoxin system encodes for mazF, which acts as an endoribonuclease that cleaves cellular mRNAs at specific sequence motifs (ACA), and mazE, which opposes the mazF action. Our goal was to detect mazEF expression in methicillin-resistant S. aureus (MRSA) isolates compared with methicillin-sensitive S. aureus (MSSA) isolates and determine its relation to methicillin susceptibility as well as biofilm-formation. According to their susceptibility to cefoxitin disks, 100 S. aureus isolates obtained from patients admitted to Cairo University Hospitals were categorized into 50 MSSA and 50 MRSA according to their susceptibility to cefoxitin disks (30 µg). Antimicrobial susceptibility and biofilm-formation were investigated using the disk diffusion method and tissue culture plate method, respectively. Finally, using real-time PCR, mazEF expression was estimated and correlated to methicillin susceptibility and biofilm formation. Both MRSA and MSSA isolates showed the best sensitivity results with linezolid and gentamicin, where about 88% of MRSA isolates and 96% of MSSA isolates were sensitive to linezolid while 76% of MRSA isolates and 84% of MSSA isolates were sensitive to gentamicin. MRSA isolates were significantly more able to form biofilm than MSSA isolates (p-value = 0.037). The mazEF expression was significantly correlated to methicillin resistance in S. aureus (p-value < 0.001), but not to biofilm-formation.


2004 ◽  
Vol 48 (4) ◽  
pp. 1124-1127 ◽  
Author(s):  
Hong Bin Kim ◽  
Hee-Chang Jang ◽  
Hee Jung Nam ◽  
Yeong Seon Lee ◽  
Bong Su Kim ◽  
...  

ABSTRACT Staphylococcus aureus, one of the most frequently isolated pathogens in both hospitals and the community, has been particularly efficient at developing resistance to antimicrobial agents. As methicillin-resistant S. aureus (MRSA) has prevailed and, furthermore, as S. aureus with reduced susceptibility to vancomycin has emerged, the therapeutic options for the treatment of S. aureus infections have become limited. To update the current status of antibiotic resistance, clinical S. aureus isolates were collected from eight university-affiliated hospitals from June 1999 to January 2001. Susceptibility tests with 28 antibiotics were performed by the disk diffusion method. Among a total of 682 isolates, the methicillin resistance rate was 64% (439 of 682), and most of the MRSA isolates were resistant to multiple classes of antibiotics. Although a constitutive macrolide-lincosamide-streptogramin B resistance phenotype was common, no isolates were resistant to quinupristin-dalfopristin or linezolid. Rifampin, fusidic acid, trimethoprim-sulfamethoxazole, and arbekacin showed superior in vitro activity compared with the other antibiotics against the MRSA isolates. No isolates showed reduced susceptibility to vancomycin.


Materials ◽  
2021 ◽  
Vol 14 (16) ◽  
pp. 4603
Author(s):  
Andrej Pavlic ◽  
Gabrijela Begic ◽  
Marin Tota ◽  
Maja Abram ◽  
Stjepan Spalj ◽  
...  

The presence of nickel could modify bacterial behavior and susceptibility to antimicrobial agents. Adhesion and biofilm formation on orthodontic archwires can be a source of bacterial colonization and possible health hazards. Staphylococcus aureus was subjected to exposure and adaptation to various sub-inhibitory concentrations of nickel. Five strains of bacteria adapted to nickel in concentrations of 62.5–1000 μg/mL were tested for adhesion and biofilm formation on nickel-titanium archwires. Archwires were previously incubated in artificial saliva. Bacteria were incubated with orthodontic wire with stirring for 4 h (adhesion) and 24 h (biofilm formation). The number of adherent bacteria was determined after sonication and cultivation on the Muller-Hinton agar. Disk diffusion method was performed on all bacteria to assess the differences in antimicrobial susceptibility. Bacteria adapted to lower concentrations of nickel adhered better to nickel-titanium than strains adapted to higher concentrations of nickel (p < 0.05). Biofilm formation was highest in strains adapted to 250 and 500 μg/mL of nickel (p < 0.05). The highest biofilm biomass was measured for strains adapted to 250 μg/mL, followed by those adapted to 1000 μg/mL. Bacteria adapted to lower concentrations of nickel demonstrated lower inhibition zone diameters in the disk diffusion method (p < 0.05), indicating increased antimicrobial resistance. In conclusion, bacteria adapted to 250 μg/mL of nickel ions adhered better, demonstrated higher biofilm formation and often had higher antimicrobial resistance than other adapted and non-adapted strains.


2020 ◽  
Vol 13 (12) ◽  
pp. 2736-2742
Author(s):  
Matlale Phriskey Mphahlele ◽  
James Wabwire Oguttu ◽  
Inge-Marie Petzer ◽  
Daniel Nenene Qekwana

Background and Aim: Staphylococcus aureus infections and antimicrobial resistance (AMR) in mastitis cases are both of clinical and economic importance. This study investigated the prevalence and AMR patterns of S. aureus isolated from composite milk samples of dairy cows submitted to the Onderstepoort Milk Laboratory for routine diagnosis. Materials and Methods: A total of 2862 cow milk samples randomly selected from submitted samples were tested for the presence of S. aureus using microbiological and biochemical tests. Confirmation of isolates was done using the analytical profile index. Antimicrobial susceptibility of S. aureus isolates against 12 antimicrobial agents was determined using the disk diffusion method. Results: S. aureus was isolated from 1.7% (50/2862) of the samples tested. All (100%) S. aureus isolates were resistant to at least one antimicrobial, while 62% (31/50) were resistant to three or more categories of antimicrobials (multidrug-resistant [MDR]). Most S. aureus isolates were resistant to erythromycin (62%; 31/50) and ampicillin (62%; 31/50). Almost half of S. aureus isolates were resistant to oxacillin (46%; 23/50) and only 8% (4/50) were resistant to cefoxitin. Conclusion: Although the prevalence of S. aureus among mastitis cases in this study was low, isolates exhibited high resistance to aminoglycosides, macrolides, and penicillins, all of which are important drugs in human medicine. The high prevalence of MDR S. aureus and the presence of methicillin resistance among S. aureus observed in this study are of both clinical and public health concerns.


Author(s):  
Harsha Sreedharan ◽  
KB Asha Pai

Introduction: Methicillin-Resistant Staphylococcus aureus(MRSA) infection is a major global healthcare problem, the prevalence of which varies from 25-50% in India. It is known to cause Skin and Soft tissue Infections (SSI), endovascular infections, endocarditis, pneumonia, septic arthritis, osteomyelitis, and sepsis. Vancomycin is the drug of choice for treating severe MRSA infections. Ceftaroline, a fifth-generation cephalosporin has been approved by the United States Food and Drug Administration (US FDA) for treating acute bacterial SSI caused by susceptible micro-organisms including MRSA, Community acquired respiratory tract infection, MRSA bacteremia and endocarditis. Aim: To assess the susceptibility of clinical isolates of S. aureusto ceftaroline, in a Tertiary Care Hospital. Materials and Methods: This prospective study was conducted in the Department of Microbiology of a Tertiary Care Hospital over a period of two months from June 2019 to July 2019. S.aureus isolates from various clinical samples were screened for methicillin resistance by disc diffusion method using cefoxitin disc and ceftaroline susceptibility of these isolates was assessed by E-strip method. The isolates were classified as ceftaroline susceptible, Susceptibility Dose Dependent (SDD) and ceftaroline resistant respectively as per CLSI guidelines. A descriptive analysis of the data was done and the results were presented as frequencies and percentages. Results: All the S.aureus isolates were found to be susceptible to ceftaroline. Methicillin Sensitive Staphylococcus aureus(MSSA) isolates had lower Minimum Inhibitory Concentration (MIC) when compared to MRSA. The highest MIC among MRSA was 0.5 μg/mL. Conclusion: Ceftaroline can be considered as an effective alternative for treatment of infections caused by MRSA.


2021 ◽  
Author(s):  
Hossein Jafari Soghondicolaei ◽  
Mohammad Ahanjan ◽  
Mehrdad Gholami ◽  
Bahman Mirzaei ◽  
Hamid Reza Goli

Abstract Biofilm production increases Staphylococcus aureus resistance to antibiotics and also host defense mechanisms. The current study aims to evaluate the biofilm formation by S. aureus and to determine the prevalence of fibronectin-binding protein genes, also its correlation with drug resistance. In this study, 100 clinical isolates of S. aureus were collected. The antibiotic susceptibility pattern of the isolates was evaluated by the disk agar diffusion method. The ability of biofilm formation in the studied isolates was also determined by microplate colorimetric assay. Then, all isolates were screened by polymerase chain reaction for the fnbA and fnbB genes. Out of 100 clinical isolates of S. aureus, the highest and lowest antibiotic resistance rates were against penicillin (94%) and vancomycin (6%). Thirty-two cases were found to be multi-drug resistant (MDR) among the all strains. The ability of biofilm production was observed in 89% of the isolates. The PCR results showed that the prevalence of fnbA and fnbB genes were 91% and 17%, respectively. Moreover, 100% and 21.8% of the MDR strains harbored the fnbA and fnbB genes respectively. The ability to form biofilm in MDR isolates of S. aureus is more than non-MDR isolates, especially fnbA positive ones. As the bacteria in the biofilm are difficult to kill by antibiotics, attention to the removal or control of the biofilm production seems to be necessary.


Author(s):  
Fateme DAVARZANI ◽  
Navid SAIDI ◽  
Saeed BESHARATI ◽  
Horieh SADERI ◽  
Iraj RASOOLI ◽  
...  

Background: Pseudomonas aeruginosa is one of the most common opportunistic bacteria causing nosocomial infections, which has significant resistance to antimicrobial agents. This bacterium is a biofilm and alginate producer. Biofilm increases the bacterial resistance to antibiotics and the immune system. Therefore, the present study was conducted to investigate the biofilm formation, alginate production and antimicrobial resistance patterns in the clinical isolates of P. aeruginosa. Methods: One hundred isolates of P. aeruginosa were collected during the study period (from Dec 2017 to Jul 2018) from different clinical samples of the patients admitted to Milad and Pars Hospitals at Tehran, Iran. Isolates were identified and confirmed by phenotypic and genotypic methods. Antimicrobial susceptibility was specified by the disk diffusion method. Biofilm formation and alginate production were measured by microtiter plate and carbazole assay, respectively. Results: Sixteen isolates were resistant to all the 12 studied antibiotics. Moreover, 31 isolates were MultidrugResistant (MDR). The highest resistance rate was related to ofloxacin (36 isolates) and the least resistance was related to piperacillin-tazobactam (21 isolates). All the isolates could produce the biofilm and alginate. The number of isolates producing strong, medium and weak biofilms was equal to 34, 52, and 14, respectively. Alginate production was more than 400 μg/ml in 39 isolates, 250-400 μg/ml in 51 isolates and less than 250 μg/ml in 10 isolates. Conclusion: High prevalence of MDR, biofilm formation, and alginate production were observed among the clinical isolates of P. aeruginosa. The results also showed a significant relationship between the amount of alginate production and the level of biofilm formation.


Author(s):  
Maysa Serpa ◽  
Juliana Amália Fonte Bôa do Nascimento ◽  
Mirian Fátima Alves ◽  
Maria Isabel Maldonado Coelho Guedes ◽  
Adrienny Trindade Reis ◽  
...  

Antimicrobial resistance is a current and important issue to public health, and it is usually associated with the indiscriminate use of antimicrobials in animal production. This study aimed to evaluate the antimicrobial susceptibility profile in bacterial isolates from pigs with clinical respiratory signs in Brazil. One hundred sixty bacterial strains isolated from pigs from 51 pig farms in Brazil were studied. In vitro disk-diffusion method was employed using 14 antimicrobial agents: amoxicillin, penicillin, ceftiofur, ciprofloxacin, enrofloxacin, chlortetracycline, doxycycline, oxytetracycline, tetracycline, erythromycin, tilmicosin, florfenicol, lincomycin, and sulfadiazine/trimethoprim. The majority of isolates were resistant to at least one antimicrobial agent (98.75%; 158/160), while 31.25% (50/160) of the strains were multidrug resistant. Streptococcus suis and Bordetella bronchiseptica were the pathogens that showed higher resistance levels. Haemophilus parasuis showed high resistance levels to sulfadiazine/trimethoprim (9/18=50%). We observed that isolates from the midwestern and southern regions exhibited four times greater chance of being multidrug resistant than the isolates from the southeastern region studied. Overall, the results of the present study showed a great level of resistance to lincomycin, erythromycin, sulfadiazine/trimethoprim, and tetracycline among bacterial respiratory pathogens isolated from pigs in Brazil. The high levels of antimicrobial resistance in swine respiratory bacterial pathogens highlight the need for the proper use of antimicrobials in Brazilian pig farms.


Antibiotics ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 375
Author(s):  
Vanessa Silva ◽  
Sara Hermenegildo ◽  
Catarina Ferreira ◽  
Célia M. Manaia ◽  
Rosa Capita ◽  
...  

In this study we aimed to characterize antimicrobial resistance in methicillin-resistant Staphylococcus aureus (MRSA) isolated from bloodstream infections as well as the associated genetic lineages of the isolates. Sixteen MRSA isolates were recovered from bacteremia samples from inpatients between 2016 and 2019. The antimicrobial susceptibility of these isolates was tested by the Kirby–Bauer disk diffusion method against 14 antimicrobial agents. To determine the macrolide–lincosamide–streptogramin B (MLSB) resistance phenotype of the isolates, erythromycin-resistant isolates were assessed by double-disk diffusion (D-test). The resistance and virulence genes were screened by polymerase chain reaction (PCR). All isolates were characterized by multilocus sequence typing (MLST), spa typing, staphylococcal chromosomal cassette mec (SCCmec) typing, and accessory gene regulator (agr) typing. Isolates showed resistance to cefoxitin, penicillin, ciprofloxacin, erythromycin, fusidic acid, clindamycin, and aminoglycosides, confirmed by the presence of the blaZ, ermA, ermC, mphC, msrA/B, aac(6’)-Ie-aph(2’’)-Ia, and ant(4’)-Ia genes. Three isolates were Panton–Valentine-leukocidin-positive. Most strains (n = 12) presented an inducible MLSB phenotype. The isolates were ascribed to eight spa-types (t747, t002, t020, t1084, t008, t10682, t18526, and t1370) and four MLSTs (ST22, ST5, ST105, and ST8). Overall, most (n = 12) MRSA isolates had a multidrug-resistance profile with inducible MLSB phenotypes and belonged to epidemic MRSA clones.


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