Μg Disc Research Articles

Investigation of the synergistic effect ofceftazidime-avibactam and aztreonam combination on carbapenem-resistant Klebsiella pneumoniae isolates with 3 different methods.

Treatment options are limited for infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates due to the production of metallo-β-lactamase (MBL). The ceftazidime-avibactam (CZA)/ aztreonam (ATM) combination represents a new therapeutic approach in MBL-positive isolates. Our study aims to determine distribution of carbapenemase genes in CRKP isolates and to investigate the in vitro synergistic effect of the CZA/ATM combination.Our study included 48 CRKP strains isolated from various clinical samples. Identification was performed using MALDI-TOF MS (bioMérieux, France), and susceptibility was tested with Vitek-2 (bioMérieux). The susceptibility to CZA and ATM was determined using CZA 30/20 µg and ATM 30 µg (Oxoid™,UK) disks. Carbapenemase genes VIM, NDM, IMP, KPC, OXA-23, OXA-58, OXA-48, and OXA-51 were investigated in only 44 isolates using the Bio-Speedy Carbapenem resistance qPCR (Bioexen, Turkiye) kit. Synergy testing was evaluated with double disk diffusion, gradient strip (bioMérieux)/disk diffusion, and broth disk elution methods.Out of 48 carbapenem-resistant isolates, 40 (83.3%) isolates showed resistance to CZA and 46 (95.8%) to aztreonam. Synergy was detected with all three methods in all isolates identified as resistant to CZA, CZA-sensitive isolates were not included in this evaluation. The most frequently detected carbapenemase genes were NDM+OXA-48, found in 28 (63.6%) of the isolates.Although the NDM+OXA-48 coexistence predominates in our center, in vitro synergy between CZA and ATM was detected in all of CZA-resistant isolates. Performing the CZA+ATM synergy test and reporting the result is crucial for choosing appropriate treatment in CRKP infection.

Assessment of lefamulin 20 µg disk versus broth microdilution when tested against common respiratory pathogens

ObjectiveTo evaluate the performance of the disk diffusion test with lefamulin 20 µg compared with the Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution (BMD) method. MethodsA total of 572 clinical stains, including 240 Staphylococcus aureus, 211 Streptococcus pneumoniae, and 121 Haemophilus influenzae, isolated from 71 medical centres from the China Antimicrobial Surveillance Network in 2020. BMD method and disk diffusion methods were performed according to CLSI. Categorical agreement (CA), major error (ME), and very ME (VME) were calculated. ResultsLefamulin showed potent activity against S. aureus, S. pneumoniae, and H. influenzae. Using the BMD method, lefamulin inhibited 97.1% of S. aureus isolates at 0.25 mg/L; seven isolates were not susceptible. For S. pneumoniae and H. influenzae, the percentage of susceptibility to lefamulin was 100% and no non-susceptible strains were found in this study. Compared with the reference BMD method, the CA of the lefamulin 20 µg disk testing was 99.8% (571/572), with 14.3% (1/7) VME and no ME. In our study, VME was determined in S. aureus. For S. pneumoniae and H. influenzae, the VME was not determined due to the lack of lefamulin non-susceptible strains. ConclusionsThe lefamulin 20 µg disk diffusion testing showed excellent CA and ME with the reference BMD method for S. aureus, S. pneumoniae, and H. influenzae. The VME exceeding CLSI recommendations may be a bias due to fewer lefamulin non-susceptible isolates. Our results suggest that lefamulin non-susceptible isolates detected by disk diffusion should be confirmed by the reference BMD.

Real-world performance of susceptibility testing for cefiderocol: insights from a prospective multicentre study on Gram-negative bacteria.

Cefiderocol is a novel siderophore-conjugated cephalosporin developed for the treatment of multidrug-resistant Gram-negative bacterial (GNB) infections. However, the current gold standard for cefiderocol susceptibility testing, broth microdilution (BMD) using iron-depleted cation-adjusted Mueller-Hinton broth, presents challenges for many microbiology laboratories. In this study, we evaluate the real-world performance of disc diffusion (DD) and a commercial BMD method (ComASP®) to test cefiderocol susceptibility in a series of isolates collected prospectively from severely ill patients in a multicentre study. The susceptibilities of 1472 isolates (632 Enterobacterales, 532 Pseudomonas aeruginosa, 84 Acinetobacter spp. and 224 Stenotrophomonas maltophilia) collected in 60 Spanish hospitals were analysed following the EUCAST 2023 and 2024 criteria. We assessed the performance of DD (cefiderocol 30 μg disc, Liofilchem) and a commercial BMD method (ComASP® Cefiderocol, Liofilchem). A total of 1408 and 1450 isolates were susceptible by DD and ComASP® BMD, respectively. Overall, the agreement between both methods was 96.9%. Forty-four isolates were resistant by DD but susceptible by ComASP® BMD, and two were susceptible by DD but resistant by ComASP® BMD (Acinetobacter baumannii isolates). Adoption of the updated 2024 EUCAST DD breakpoints and areas of technical uncertainty (ATUs) led to a decrease in susceptibility among Enterobacterales (95.3% versus 92.6%). DD is a straightforward, rapid and accessible method for routine determination of cefiderocol susceptibility in real-world practice. ComASP® BMD shows a high agreement with DD in susceptible isolates and may help to resolve DD interpretability concerns in isolates with susceptibility results within the ATU, but caution is warranted when testing resistant isolates.

Surveillance of In Vitro Activity of Cefiderocol Against Carbapenem-Resistant Gram-Negative Clinical Isolates in a Tertiary Care Hospital.

Antibiotic resistance among Gram-negative bacterial isolates is increasingly observed. With the emergence of carbapenem-resistant and pan-resistant pathogens, treating these resistant infections is becoming more challenging due to the limited number of effective drugs. There is a desperate need for the discovery of new antibiotics with novel mechanisms of action. Cefiderocol is one such novel antibiotic with a unique siderophore-based mechanism of action, which has been recently approved for clinical use against drug-resistant pathogens. The present study aims to identify the in vitro activity of cefiderocol against major carbapenem-resistant clinical isolates, including those resistant to colistin. One hundred and one carbapenem-resistant clinical isolates were included in the study. Identification and antibiotic susceptibility testing were performed using the automated VITEK® 2 Compact (bioMérieux SA, Marcy-l'Étoile, France) identification and susceptibility testing system, except for colistin and cefiderocol. Colistin resistance in Enterobacterales and Pseudomonas aeruginosa was assessed using the agar dilution minimum inhibitory concentration method, while for Acinetobacter baumannii, broth microdilution method was employed. Cefiderocol susceptibility testing was conducted using the Kirby-Bauer disc diffusion method with 30 µg discs on standard Mueller-Hinton agar plates. For selected isolates, cefiderocol minimum inhibitory concentration detection was performed using broth microdilution with iron-depleted cation-adjusted Mueller-Hinton broth. Of the total 101 isolates, the majority (75, 74.25%) were Enterobacterales which includedKlebsiella pneumonia (42, 41.58%) and Escherichia coli (33, 32.67%), followed by Pseudomonas aeruginosa (13, 12.87%) and Acinetobacter baumannii (10, 9.9%). Only three (2.97%)of the isolates were Stenotrophomonas maltophilia. Most of the isolates were susceptible to cefiderocol, with only four (3.96%) isolates showing resistance. Colistin resistance was observed in six (6.12%) of the isolates. There was a good correlation between disc diffusion testing and broth microdilution testing for the detection of cefiderocol-resistant isolates. No cross-resistance with colistin was observed, as all colistin-resistant isolates were uniformly susceptible to cefiderocol Conclusion: Cefiderocol is highly effective with good in vitro activity against the majority of carbapenem-resistant pathogens. While some isolates do show resistance, it is relatively uncommon. Given its safety profile compared to colistin, cefiderocol can serve as an alternative to colistin to treat carbapenem-resistant infections and it may be considered even for the management of colistin-resistant cases. Disc diffusion testing is a reliable method for identifying cefiderocol-resistant isolates in routine clinical and diagnostic laboratories, especially in resource-limited settings.

Current trend of biapenem susceptibility and disc diffusion breakpoints in Enterobacterales and Pseudomonas aeruginosa

IntroductionBiapenem has been recently approved by the Drug Controller General of India for the treatment of complicated urinary tract infections (cUTI). However, there are no assessment studies that evaluate the in-vitro activity of biapenem against contemporary ESBL-producing Indian Enterobacterales isolates. To determine the activity of biapenem against contemporary ESBLs and/or OXA-1/ampC producing Enterobacterales and Pseudomonas aeruginosa isolates. MethodologyIsolates were tested for susceptibility to biapenem and its comparators using the broth microdilution method. Presence of ESBLs (SHV, TEM, CTX-M) genes, OXA-1, and ampC genes (ACC, ACT, DHA, CIT/CMY, FOX) using multiplex PCR. ResultsAgainst ESBL with OXA-1 and/or ampC-producing E. coli, ESBL-K. pneumoniae, and cephalosporin-resistant P. aeruginosa, biapenem showed in-vitro activity similar to that of meropenem. Overall, a biapenem disc concentration of 10 μg provided no error rates for testing E. coli, K. pneumoniae, and P. aeruginosa isolates. ConclusionIt is more accurate to test biapenem at a 10 μg disc concentration and apply more stringent disc diffusion breakpoints for interpretation.

Performance of modified colistin broth disc elution vis-a-vis broth microdilution method for susceptibility testing of Enterobacterales

Objectives: Recently, Clinical and Laboratory Standards Institute (CLSI) has approved colistin broth disc elution (CBDE) to be a supplemental test. This requires multiple discs and tubes to get the desired concentrations of colistin -1, 2, and 4 µg/mL and 10 mL volume of cation-adjusted Mueller–Hinton broth for a single isolate. The present study was aimed to evaluate the performance of CBDE in a microtiter plate format modified (mCBDE) with the reference method broth microdilution (BMD) for detection of colistin resistance in carbapenem-resistant Enterobacterales (CRE) isolates. Materials and Methods: One hundred and sixty non-duplicate clinical CRE isolates (May 2021–April 2022) were simultaneously subjected for BMD and mCBDE. For mCBDE, colistin 10 µg discs and Mueller–Hinton broth no-2 control cations were procured from HiMedia, Mumbai, and drug concentrations were prepared following CLSI-M100Ed31. Results of mCBDE were compared with reference BMD (Minimum inhibitory concentration [MIC] ≤2 µg/mL – intermediate and ≥4 µg/mL – resistant). Statistical Analysis: The performance of mCBDE was compared with BMD and expressed in terms of Categorical, essential agreement (EA), very major error (VME), and major error (ME). The sensitivity and specificity were calculated using Fisher’s contingency Table. Results: Of the 160 CRE isolates, 152 had exactly the same minimal inhibitory concentration (MIC) in both the tests with four isolates having higher and four having lower colistin MIC by mCBDE, giving a major error of 2.1% and VME of 5.5%. Categorical and essential agreement of mCBDE were 97.5% and 98.7%, respectively. Conclusions: mCBDE is an easy, economical, and reliable alternative test for determining colistin susceptibility for CRE isolates. Further, large-scale study is needed to strengthen our observation.

Detection of blaKPC gene among carbapenemase producing Klebsiella pneumoniae isolated from different clinical specimens at tertiary care hospital of Nepal

BackgroundKlebsiella pneumoniae infections have become a major cause of hospital acquired infection worldwide with the increased rate of acquisition of resistance to antibiotics. Carbapenem resistance mainly among Gram negative is an ongoing problem which causes serious outbreaks dramatically limiting treatment options. This prospective cross-sectional study was designed to detect blaKPC gene from carbapenem resistant K. pneumoniae.Materials and MethodsA totally of 1118 different clinical specimens were screened and confirmed for KPC producing K. pneumoniae phenotypically using Meropenem (10 μg) disc. The blaKPC gene was amplified from the isolates of K. pneumoniae to detect the presence of this gene.ResultOf the total samples processed, 18.6% (n = 36) were K. pneumoniae and among 36 K. pneumoniae, 61.1% (n = 22/36) were meropenem resistant. This study demonstrated the higher level of MDR 91.7% (n = 33) and KPC production 47.2% (n = 17) among K. pneumoniae isolates. The blaKPC gene was detected in 8.3% (n = 3) of meropenem resistant isolates.ConclusionSince the study demonstrates the higher level of MDR and KPC producing K. pneumoniae isolates that has challenged the use of antimicrobial agents, continuous microbiology, and molecular surveillance to assist early detection and minimize the further dissemination of blaKPC should be initiated. We anticipate that the findings of this study will be useful in understanding the prevalence of KPC-producing K. pneumoniae in Nepal.

Performance of disk diffusion method for aztreonam in combination with avibactam against Enterobacteriales.

To evaluate the performance of an in-house developed disk diffusion method for aztreonam in combination with avibactam against Enterobacteriales. The in vitro antibacterial activity of aztreonam with avibactam against 204 carbapenemase-producing Enterobacteriales was determined by a disk diffusion method, with a broth microdilution method as a reference. The optimal S/R breakpoints for disk diffusion tests of 30/20 and 10/4 µg disks, calculated by the dBETs software using the model-based approaches, were ≥22/≤21 and ≥12/≤11 mm, respectively. On the basis of the estimated breakpoints, the CAs for disk diffusion tests of 30/20 and 10/4 µg aztreonam/avibactam disks were both 98.0%, with 0.5% major error and 37.5% very major error. The home-made disk diffusion method is an economical and practical method for clinical microbiology laboratories to determine the antibacterial susceptibility of aztreonam with avibactam against Enterobacteriales.

COMPARISON OF NASAL CARRIAGE OF MRSA AND ITS ANTIBIOTIC SUSCEPTIBILITY WITH SPECIAL REFERENCE TO MUPIROCIN AMONG STUDENTS OF HEALTH-CARE SETTINGS AND OTHER STUDENTS

Objectives: One of the potential risk factors for nosocomial staphylococcal and Methicillin-resistant Staphylococcus aureus (MRSA) infections is colonization of the anterior nares of HCWs, and from there they may spread to the patients, which may cause a burden on the health-care system. In this context, we made an attempt to compare the Staphylococcal and MRSA nasal carriage and their susceptibility to Mupirocin in students who are exposed to the health-care system and those who are not. Methods: Nasal swabs were collected from 100 paramedical students as the study group and 100 non-medical students as the control group, all in the age group between 18 and 21 years. 5% sheep blood agar and mannitol salt agar were used to isolate S. aureus, and antibiotic sensitivity was done by Kirby Bauer Disc Diffusion Technique. Cefoxitin disc (30 μg) and Vancomycin screen agar were used to detect MRSA and Vancomycin resistance, respectively. A 5 μg disc of Mupirocin was used to test the susceptibility of Mupirocin. Results: Nasal carriage rate of S. aureus and MRSA was 23% and 8% in Paramedical students and 17% and 4% in Nonmedical students respectively. 83% of isolates from paraclinical students and 95% of isolates from non-medical students were susceptible to linezolid. Out of 12 MRSA strains from both groups only one strain showed resistance to Mupirocin. Conclusions: Nasal carriage of S. aureus and MRSA and antibiotic resistance of isolated strains were more common in paramedical students and also in female students. Vancomycin resistance was not observed in MRSA strains from male students. Mupirocin resistance, even in a single case, needs to be addressed.

Study of Methicillin-resistant Staphylococcus aureus Isolated from Various Clinical Specimens from Tertiary Care Hospital

ABSTRACT Introduction: Staphylococci are Gram-positive bacteria, with a diameter of 0.5–1.5 μm and individual cocci, which are divided in more than one plane from the grape-like cluster. In the genus staphylococci, 32 species and 8 subspecies are present in many of which are present as a commensal in the human body. Staphylococci are frequently found as a normal flora of skin mucosal, surface of respiratory, upper elementary, and urogenital tract of the human body. In immunocompromised condition, the infection can be spread by skin-to-skin contact and also by excretion such as saliva aerosol releasing during sneezing and coughing. Infection of staphylococci may also spread from the animal product such as nonpasteurized milk and milk products. Over the last few decades, Staphylococcus aureus has emerge as the most dangerous organism, which is known as methicillin-resistant S. aureus (MRSA). Some strains show vancomycin resistance by S. aureus in hospitalized patients. The present study was conducted to study the prevalence and antibiogram of MRSA isolates in a clinical sample from Acharya Vinoba Bhave Rural Hospital. Materials and Methods: This study was conducted in the Department of Microbiology at Jawaharlal Nehru Medical College (Datta Meghe Institute of Higher Education and Research), Wardha, from August 2020 to December 2021. The samples received in the microbiology laboratory were inoculated on blood agar and MacConkey’s agar, and incubated for 24 h at 37°C for staphylococcal isolation. A total of 145 Staphylococcal aureus were isolated and confirmed by Gram stain and various biochemical tests such as catalase test, oxidase test, coagulase test, and mannitol salt agar. Antibiotic susceptibility test was examined on the Mueller–Hinton agar plate using the modified Kirby–Bauer disc diffusion method. Screening of methicillin resistant was performed by cefoxitin (30 μg) disc diffusion method and interpreted according to CLSI guideline. Results: The prevalence of MRSA was 76 (52.41%) to methicillin-susceptible S. aureus. The highest MRSA was found in the age group >50 years (26.31%). Out of 76 MRSA isolates from different departments, the medicine and orthopedic departments show maximum MRSA as 19.74% each followed by pediatrics (17.11%) and surgery (13.16). Out of 76 MRSA isolates, vancomycin showed 93.42% sensitivity followed by linezolid (85.52%), Gentamicin (71.05%), tetracycline (56.57%), clindamycin (46.05%), and erythromycin (31.57%). Conclusion: With the advancement of new therapeutic agent, MRSA is also being easily resistant. Due to increasing cases of MRSA, there is an increase in the rate of morbidity and mortality. Moreover, except vancomycin, other drugs are resistant, as their MIC is not in the susceptible range. Thus, the results of this study indicate that vancomycin is a suitable medication to take when MRSA cases rise. In such cases, MRSA can be controlled by regular monitoring and checking the irrational use of antibiotics.

Clindamycin Resistance Detection among MRSA at a Tertiary Care Hospital: A Cross-sectional Study

Introduction: Antimicrobial resistance is on the rise, posing a global threat to the success of modern surgical procedures. A common and notorious superbug causing community and hospital-acquired infections is Methicillin-resistant Staphylococcus aureus (MRSA). Its potential for easy and rapid spread leads to increased mortality in hospitals. Clindamycin, a relatively inexpensive and effective drug, is empirically prescribed for the treatment of MRSA infections. Due to the risk of misidentifying clindamycin resistance as susceptibility, leading to treatment failure, its judicious use after D-test is advocated. Aim: To isolate Staphylococcus aureus (S.aureus) from various clinical specimens and to identify MRSA using cefoxitin disc (30 μg), further determining the incidence of inducible clindamycin resistance among MRSA by the disc diffusion method (D-test). Materials and Methods: The present cross-sectional study was conducted in the Department of Microbiology, Government Mohan Kumaramangalam Medical College Hospital (GMKMCH), Salem, Tamil Nadu, India, from June 2023 to August 2023. Study was conducted using 185 S.aureus isolates collected from samples received from patients of all genders treated at the present study hospital. Various clinical samples such as pus, sputum, blood, urine and fluids collected from patients treated at the study Institute Salem during the study period were included. A cefoxitin disc (30 μg) was used to detect MRSA among the 185 S.aureus isolates by disc diffusion method as per Clinical and Laboratory Standards Institute (CLSI) guidelines, which were further tested for clindamycin resistance using clindamycin (2 μg) with erythromycin (15 μg) discs by the D-test. The data of the study were analysed using Statistical Package for the Social Sciences (SPSS) software version 21.0. Further descriptive statistics and a chi-square test with a level of significance, p-value ≤0.005 (statistically significant), were used. Results: Among 185 S.aureus isolates, 130 (70.27%) were Methicillin-sensitive S.aureus (MSSA) and 55 (29.7%) were MRSA. Inducible clindamycin resistance was observed in 22 (40%) isolates, while 17 (30.9%) isolates showed constitutive resistance, 9 (16.36%) showed the MS phenotype, and the remaining 7 (12.72%) showed the susceptible phenotype. Among the clindamycin resistance patterns in MRSA, inducible clindamycin resistance was reported predominantly. Conclusion: The majority of S.aureus was isolated from pus samples, highlighting its importance as a pyogenic microorganism. The current study records a high rate of MRSA resistance among S.aureus isolates. The present study reports a higher rate of inducible clindamycin resistance among MRSA isolates when compared to other phenotypes of clindamycin resistance. Therefore, routine D-testing must be implemented to identify inducible resistance to clindamycin in MRSA to avoid treatment failure.

Usefulness of inclusion of ertapenem and temocillin screening breakpoints in the EUCAST rapid antimicrobial susceptibility testing (RAST) for rapid detection of OXA-48-producing Klebsiella pneumoniae directly from positive blood cultures.

The aims of this study were: (i) to assess the ability of the meropenem screening breakpoint as part of the screening rapid antimicrobial susceptibility testing (sRAST) of EUCAST for the detection of OXA-48 carbapenemase-producing Klebsiella pneumoniae directly from positive blood cultures (BCs); and (ii) to evaluate the inclusion of ertapenem and temocillin discs into the sRAST to enhance the detection of OXA-48-producing isolates. BC bottles were spiked with a total of 117 K. pneumoniae isolates, including 77 previously characterized OXA-48 producers and 40 non-OXA-48 producers. Disc diffusion assays were directly performed from positive BCs with meropenem (10 µg), ertapenem (10 µg) and temocillin (30 µg) discs, and inhibition zones were manually measured after 4, 6 and 8 h of incubation. The screening cut-off values of sRAST were applied to evaluate their capability in detecting OXA-48-producing isolates. Receiver operating characteristic curves were constructed to illustrate the performance efficacy of the disc diffusion assays to detect OXA-48 producers. The meropenem cut-off values of sRAST only detected 90.91% of the OXA-48-producing isolates after 6 and 8 h of incubation. With the proposed cut-off points for ertapenem [<19 mm (4/6 h) and <20 mm (8 h)] and temocillin [<10 mm (4 h) and <11 mm (6/8 h)], all OXA-48-positive isolates were detected without any false-positive results at any reading time. In healthcare settings with a high prevalence of OXA-48 producers, the inclusion of ertapenem and temocillin discs in the sRAST procedure may improve the detection of OXA-48-producing K. pneumoniae isolates directly from positive BCs, providing reliable results after only a 4 h incubation period.

Sulbactam-durlobactam susceptibility test method development and quality control ranges for MIC and disk diffusion tests.

Sulbactam-durlobactam is a β-lactam/β-lactamase inhibitor combination developed to treat hospital-acquired and ventilator-associated bacterial pneumonia caused by Acinetobacter baumannii-calcoaceticus complex (ABC). Durlobactam is a diazabicyclooctane β-lactamase inhibitor with potent activity against Ambler classes A, C, and D serine β-lactamases and restores sulbactam activity against multidrug-resistant ABC. Studies were conducted to establish sulbactam-durlobactam antimicrobial susceptibility testing methods for both broth microdilution minimal inhibitory concentration (MIC) and disk diffusion tests as well as quality control (QC) ranges. To establish the MIC test method, combinations of sulbactam and durlobactam were evaluated using a panel of genetically characterized A. baumannii isolates which were categorized as predicted to be susceptible or resistant based on the spectrum of β-lactamase inhibition by durlobactam. MIC testing with doubling dilutions of sulbactam with a fixed concentration of 4 µg/mL of durlobactam resulted in the greatest discrimination of the pre-defined susceptible and resistant strains. Similarly, the sulbactam/durlobactam 10/10 µg disk concentration showed the best discrimination as well as correlation with the MIC test. A. baumannii NCTC 13304 was selected for QC purposes because it assesses the activity of both sulbactam and durlobactam with clear endpoints. Multi-laboratory QC studies were conducted according to CLSI M23 Tier 2 criteria. A sulbactam-durlobactam broth MIC QC range of 0.5/4-2/4 µg/mL and a zone diameter QC range of 24-30 mm were determined for A. baumannii NCTC 13304 and have been approved by CLSI. These studies will enable clinical laboratories to perform susceptibility tests with accurate and reproducible methods.

Plasmid-Mediated AmpC (pAmpC) Genotypes Among Uropathogenic Escherichia coli: A Hospital-Based Study From Western Uttar Pradesh.

Introduction Resistance due to AmpC and extended-spectrum beta (β)-lactamases (ESBLs) in Escherichia coli is an emerging problem worldwide. AmpC enzymes are a subclass of β-lactamases that have a capacity to hydrolyze and deactivate a large range of β-lactam antibiotics, particularly cephalosporins, penicillins, and monobactams, although frequently being susceptible to carbapenems and fourth-generation cephalosporins. The prevalence of plasmid-mediated AmpC (pAmpC) genotypes in uropathogenic E. coli isolates were looked at a tertiary care teaching hospital of Western Uttar Pradesh. Materials and methods A total of 312 non-repeat clinical E. coli isolates among patients presented with urinary tract infections (UTIs) were investigated by standard microbiological methods. Isolates were screened for the presence of ampC using a cefoxitin (30 µg) disc and confirmed using an inhibitor-based assay. Using multiplex polymerase chain reaction (PCR), six AmpC genotypes, namely, CIT, DHA, EBC, ACC, FOX, and MOX, were genotypically identified. Results A total of 152 (48.72%) uropathogenic E. coliisolates tested positive on the cefoxitin screening. Out of which, AmpC production was confirmed in 118/152 (77.63%) using a phenotypic method. In particular, the pAmpC gene was found in 56/152 (36.84%) isolates. CIT was the most common gene detected in this geographical area (57.14 %). Multiple genes, i.e., CIT and FOX, were also detected in 14.29%of the isolates. Conclusion Identifying AmpC producers is important in routine microbiology laboratory as they are a nosocomial threat requiring strict adherence to infection control protocols. A confirmatory phenotypic test followed by genotypic tests will help in the correct and accurate identification of this resistance.

Sustainable one-pot synthesis of novel soluble cellulose-based nonionic biopolymers for natural antimicrobial materials

Novel soluble cellulose-based nonionic biopolymers (CIs) with enhanced antimicrobial properties and nonleachability were successfully produced using a sustainable one-pot synthesis method. Fourier transform infrared spectroscopy (FTIR), Nuclear magnetic resonance H-spectrometer (1H NMR), X-ray Photoelectron Spectroscopy (XPS), and Energy Dispersive Spectroscopy (EDS) results demonstrated that the cellulose (MCC) molecules combined with indole-3-acetic acid (IAA) via esterification to produce CIs with abundant terminal indole groups. The degree of substitution of the prepared CI3 reached 1.85 when the molar ratio of IAA to MCC molecules was 4:1. The prepared CI samples were characterized using X-ray diffractometer (XRD), Scanning Electronic Microscopy (SEM), Thermogravimetric Analysis (TGA), and other analysis techniques. Results indicated that after the MCC was grafted with IAA, its crystallinity decreased and solubility increased. After blending CI3 with polycaprolactone (PCL) to form cellulose-based antimicrobial (PCL–CI) films, the films showed good compatibility, preferable biological cell activity, and low water vapor permeability. When the CI3 content was 10%, the tensile strength of the produced PCL–CI10 film reached 9.96 MPa. Moreover, the prepared PCL–CI films exhibited good nonleachability after being immersed in water for 5 d. The disk diffusion assay revealed that the CIs and PCL–CI films had good antimicrobial and bactericidal effects against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli). The minimal inhibitory concentration was 5 μg disk−1, significantly lower than that of traditional antibiotics and chitosan. The nonionic biopolymers are simple and efficient to prepare and ecofriendly as well as exhibit nontoxicity, good solubility, enhanced antimicrobial properties, and nonleachability, which can provide new ideas for developing natural biomass-based nonionic antimicrobial materials with potential applications in wound dressing, medical devices, and food packaging.

The Emergence of Mupirocin Resistance among Staphylococcus aureus in a Tertiary Care Hospital in South India: The Necessity for Routine Susceptibility Testing

Methicillin-resistant Staphylococcus aureus (MRSA) is difficult to treat, causing considerable morbidity and mortality. Nasal carriage of MRSA can occur both in healthcare workers and patients. Mupirocin is used as a topical agent for the eradication of such isolates. The present study aims to study the prevalence of mupirocin resistance among the MRSA and MSSA (Methicillin-sensitive Staphylococcus aureus) isolates. A total of 148 Staphylococcus aureus isolates were tested. Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method for amoxicillin, penicillin, cotrimoxazole, clindamycin, mupirocin(5 µg and 200 µg discs for low and high-level resistance), erythromycin, gentamicin and linezolid. MRSA isolates were detected by cefoxitin disc diffusion and Mec A detection by PCR (Polymerase Chain Reaction). MRSA was detected among 44 (29.7%) of the isolates. Among MSSA, good susceptibility was observed for cotrimoxazole 89 (85.5%) and clindamycin 92 (88.4%). An overall mupirocin resistance of 12(8.1%) was observed, with high-level resistance at 4 (2.7%) and low-level resistance at 8 (5.4%).The mupirocin resistance pattern between MRSA and MSSA was not statistically significant (p=0.1833). The emergence of mupirocin resistance highlights the necessity for creating cognizance among clinicians before prescribing mupirocin. In eradicating nasal carriage of MRSA, all the isolates should always be tested for mupirocin susceptibility to prevent the selection and spread of drug-resistant isolates.

New indoleacetic acid-functionalized soluble oxidized starch-based nonionic biopolymers as natural antibacterial materials

This study aims to develop a new soluble oxidized starch-based nonionic antibacterial polymer (OCSI) featuring high antibacterial activity and non-leachability by grafting indoleacetic acid monomer (IAA) onto the oxidized corn starch (OCS). The synthesized OCSI was characterized analytically by Nuclear magnetic resonance H-spectrometer (1H NMR), Fourier transform infrared spectroscopy (FTIR), Ultraviolet-visible spectroscopy (UV–Vis), X-ray diffractometer (XRD), X-ray Photoelectron Spectroscopy (XPS), Scanning Electronic Microscopy (SEM), Thermogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC). The results showed that the synthesized OCSI was endowed with high thermal stability and favorable solubility, and the substitution degree reached 0.6. Besides, the disk diffusion test revealed a lowest OCSI inhibitory concentration of 5 μg disk−1, and showed significant bactericidal activity against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli). Moreover, the antibacterial films (OCSI-PCL), featuring their good compatibility, mechanical properties, antibacterial activity, non-leachability, and low water vapor permeability (WVP), were also successfully prepared by blending OCSI with biodegradable polycaprolactone (PCL). Finally, CCK-8 assay results confirmed the excellent biocompatibility of the OCSI-PCL films. Overall, this very study evidenced the applicability of the obtained oxidized starch-based biopolymers as an eco-friendly non-ionic antibacterial material and confirmed their promising applications in areas including biomedical materials, medical devices, and food packaging.

Assessing the quality of the anaerobic environment — a method developed to support EUCAST disk diffusion of anaerobic bacteria

The purpose of this study is to establish a method for assessing the anaerobic environment for EUCAST disk diffusion antimicrobial susceptibility testing (AST) of anaerobic bacteria on fastidious anaerobe agar with 5% mechanically defibrinated horse blood (FAA-HB). The method utilizes the association between a decrease in the metronidazole disk zone diameter and increasing oxygen levels with an aerotolerant Clostridium perfringens strain DSM 25589 (CCUG 75076 and NCTC 14679). The C. perfringens strain was tested on FAA-HB with a McFarland 1 inoculum and a metronidazole 5 μg disk. FAA-HB was incubated for 16–20 h at 35–37°C. The association between oxygen levels (0, 0.16, 1, 2, and 4% oxygen) and the metronidazole zone diameter was determined. Reproducibility at 0% oxygen was investigated as part of a European multi-centre study of disk diffusion of anaerobic bacteria. The median zone diameters (n=12) at each oxygen level were 29 mm (0%), 21 mm (0.16%), 16 mm (1%), 15 mm (2%), and 15 mm (4%). The metronidazole zone diameters at 0% oxygen from the multi-centre reproducibility-study had a median of 29 mm and a 95%-percentile range of 25–33 mm (n=236). Only one reading was below 25 mm. Based on our results, a zone diameter of ≥25 mm using a metronidazole 5 μg disk and the C. perfringens strain, tested with EUCAST recommendations, can be used to indicate that the anaerobic environment is of sufficient quality for culture and disk diffusion AST. EUCAST has included the method as part of the quality control for AST of anaerobic bacteria.

Comparison of Disk Diffusion and Agar Dilution Method for the Detection of Mupirocin Resistance in Staphylococcal Isolates from Skin and Soft Tissue Infections.

Aims and Objectives Mupirocin is a widely used topical antibiotic for the treatment of skin and soft tissue infections. This has resulted in resistance leading to treatment failure. Hence, the present study aimed to determine the prevalence of mupirocin resistance among staphylococcal isolates obtained from the skin and soft tissue infections. Also, comparison of disc diffusion and agar dilution method in detecting mupirocin resistance was done. Materials and Methods This cross-sectional study was conducted in the Department of Microbiology of a tertiary health care center in Karnataka from January to December, 2018. Clinical samples such as wound swabs, tissues, and pus were included in the study. All staphylococcal isolates were screened for mupirocin resistance using 5 µg and 200 µg discs for low-level (MuL) and high-level mupirocin resistance (MuH), respectively. Minimum inhibitory concentration (MIC) was determined using the agar dilution method. Results Out of 100 staphylococcal isolates, 68 were Staphylococcus aureus and 32 were CoNS. MuH was detected in 11 isolates. MuH was more common in CoNS (10/11) compared with S. aureus (1/11). MuL was not found in the study. Discussion In our study, 10 out of 11 mupirocin-resistant isolates were methicillin resistant, which is statistically significant ( p < 0.05). The correlation between results of disc diffusion and MIC were appropriate in this study. Conclusion Judicial prescription of mupirocin after knowing the susceptibility report should become the standard practice. Screening for mupirocin resistance can be done by disc diffusion in resource-limited settings.

Ampicillinase C Beta-lactamase Producers among Isolates of Enterobacteriaceae in a Tertiary Care Centre: A Descriptive Cross-sectional Study.

Ampicillinase C beta-lactamase-producing organisms are often resistant to multiple antimicrobial agents, and therapeutic options against these pathogens are limited. Limited information is available regarding Ampicillinase C beta-lactamase producers. The aim of this study was to find out the prevalence of Ampicillinase C beta-lactamase producers among isolates of Enterobacteriaceae in a tertiary care centre. A descriptive cross-sectional study was carried out in the Clinical Microbiology Laboratory of a tertiary care centre from May 2021 to October 2021. Ethical approval was received from the Institutional Review Committee (Reference number: 044-077/078). Isolates of Enterobacteriaceae from various clinical samples were collected by convenience sampling. Ampicillinase C screening for beta-lactamase producers among the Enterobacteriaceae isolates was done using cefoxitin (30 μg) disc. Detection of Ampicillinase C beta-lactamase producers among the screen-positive isolates was done by cefoxitin-cloxacillin double-disc synergy test. An increase in the zone size of ≥4 mm was considered as Ampicillinase C beta-lactamase producers. Point estimate and 95% Confidence Interval were calculated. Among the total 481 isolates of Enterobacteriaceae, 49 (10.19%) (7.50-12.90, 95 % Confidence Interval) were detected as Ampicillinase C beta-lactamase producers among isolates of Enterobacteriaceae. The prevalence of Ampicillinase C beta-lactamase producers was lower than in other studies done in similar settings. Meropenem could be a drug of choice for the treatment of infections due to Ampicillinase C beta-lactamase-producing gram-negative bacteria. antibiotic; beta-lactamase; Enterobacteriaceae; gram-negative bacteria.