Clinical Laboratory Standards Institute Research Articles

Superadded Bacterial Infections in COVID–19 Patients; Antimicrobial Susceptibility and Association with Serological Markers

Objective: To determine the frequency and antimicrobial susceptibility pattern of pathogens responsible for superadded bacterial infection in COVID-19 patients and correlate the association of these infections with serological markers.
 Study Design: Cross-sectional study.
 Place and Duration of study: Department of Pathology, Combined Military Hospital, Multan Pakistan, from Jan to Dec 2021.
 Methodology: A total of 290 patients having positive RT-PCR for SARS CoV-2 were included. All samples were processed per Clinical Laboratory Standard Institute (CLSI) protocols. API 20E and API 20NE were used for the identification of Gramnegative rods. Antimicrobial susceptibility testing was performed by the modified Kirby Bauer disc diffusion method.Serological markers, including C-reactive protein (CRP), total leucocyte count (TLC) and serum Ferritin, were determined and compared for significance in positive and negative culture cases.Results: A total of 75 patients had positive bacterial cultures. Among these, 42(56%) were blood culture, 26(35 %) were respiratory culture and 7(9%) were urine culture. Commonly isolated organisms were Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus and Pseudomonas aeruginosa, i.e., 23(31%), 20(27%), 13(17%), and 12(16%) respectively. CRP, TLC and S. ferritin were markedly raised in superadded bacterial infection compared to patients with COVID-19 infection only.
 Conclusion: The frequency of superadded bacterial infections in COVID-19 patients is high. The pathogens isolated in these cases were multidrug-resistant, reflecting mostly hospital-acquired flora. The association of serological markers in depicting superadded infection is statistically significant and may be used to screen for superadded bacterial infection in COVID-19 patients.

Essential oil composition and antimicrobial activity of Glaucosciadium cordifolium (Boiss.) Burtt. & Davis

Glaucosciadium cordifolium (Boiss.) Burtt. & Davis, which is a monotypic species naturally grown in Turkey belonging to the Apiaceae (Umbelliferae) family. In this study, the essential oil obtained from G. cordifolium was analyzed and evaluated for its antimicrobial effects. In chemical studies, the analyzes of essential oil compounds obtained from the aerial parts of plants by hydrodistillation were carried out with GC-FID and GC-MS. In total, 23 compounds were identified making up 98.7% of the total volatile constituents. Sabinene (42.1%), α-pinene (17.1%), and α-phellandrene (10.1%) were found as the main constituents in the oil. In antimicrobial studies, the anticandidal and antibacterial effects of essential oils were tested against 14 pathogenic microorganisms according to the standard protocols of the Clinical Laboratory Standards Institute (CLSI). It has been determined that essential oils have a very weak inhibitory effect when compared with standard antibacterial agents. MIC values of 1800, 3600 µg/Ml, and higher were determined. In addition, it was determined that it showed inhibitory effects, especially against C. krusei at a concentration of 250 µg/mL, with MIC values ranging from 250 to 2000 µg/mL.

PREVALENCE, ANTIMICROBIAL SUSCEPTIBILITY BACTERIURIA AMONG PREGNANT WOMEN AT SOS HOSPITAL, MUQDISHO SOMALIA


 
 
 Objective: the aim of this study is to isolate the prevalence, of antimicrobial susceptibility bacteriuria among pregnant women at SOS hospital.Methods: Descriptive cross-sectional study during the time frame between Dec 2020 at SOS hospital in Hilawa distract Muqdisho.The urine sample was collected using the midstream clean-catch urine method and tested bacteriologically following standard procedure. An antimicrobial susceptibility test was also performed for the isolated organism using the Kirby disk diffusion method according to the clinical laboratory standard institute (CLSI). 
 For data analysis and interpretation we used the SPSS tool and interpretation was presented in frequencies and percentages in tables.Result: of the 102 samples collected 34 showed no growth 68 was growth. of the 68 demonstration organisms Staphylococcus aureus 46 (41.1%). E.coli were 8 (7.8%) and pseudomonads aeruginosa 8(7.8% ) Proteus mirabiliss 3(2.9%), K.Pneumonia 3(2.9%). the most sensitive drug was Levofloxacin 51(75%). while Nitrofurantoin was the highest resistant pattern 58(85.29%) followed by Ceftriaxone 48(70.6%) amoxicillin 38(55.9%) ciprofloxacin 24 ( 35.29%) respectively. 
 
 

Cumulative antimicrobial susceptibilities for respiratory clinical isolates of Mycobacterium avium Complex, Mycobacterium kansasii, and Mycobacterium abscessus from Pakistan 2018 to 2022.

Nontuberculous mycobacteria (NTM) are increasingly identified as causes of protracted pulmonary infections. Antibiotic susceptibility testing requires microdilution methods, which are often unavailable in laboratories in resource-poor settings. We report cumulative antibiograms for the most frequently isolated clinical pulmonary NTM from Pakistan to inform empiric antibiotic management of initial NTM infections. We analyzed data from 2018 to 2022 for the most frequently isolated and clinically relevant NTM isolated from respiratory specimens, i.e., Mycobacterium avium complex (MAC), Mycobacterium abscessus group (MAG), and Mycobacterium kansasii (MK). Antibiograms were developed using the Clinical Laboratory Standards Institute's M39ED5 standard. Percentage susceptibilities and 95% confidence intervals (CI) were calculated. Over 4 years, 529 NTM, comprising 209 MAC, 249 MAG, and 71 MK were analyzed. For MAC and MAG, where clarithromycin (CLR)-based regimens are recommended, CLR was active for 94.8% (95% CI 91.3-96.9), and 77.5% (95% CI 71.4-82.7) isolates, respectively. Combination regimens comprising 3 active drugs CLR + linezolid (LZD) + moxifloxacin for MAC and CLR + LZD + Amikacin for MAG had 98.4% (95% CI 95.9-99.4) and 68.9% (95% CI 62.3-74.8) coverage for pulmonary disease, respectively. For MK, 91.5% (95% CI 82.8-96.1) isolates were susceptible to rifampin (RIF), with a combination of RIF + CLR covering 88.7% (95% CI 79.3-94.2) of MK pulmonary infections, respectively. These data can inform empiric treatment guidance for the most common NTM pulmonary infections, i.e., for MAC, MAG, and MK disease in Pakistan.

Ciprofloxacin resistance and tolerance of Pseudomonas aeruginosa ocular isolates

PurposeTolerance to antibiotics may occur due to changes in bacterial growth patterns and can be a precursor to development of resistance. However, there is a lack of information on the ability of ocular bacteria isolates to develop tolerance. This paper explores the tolerance to 8 different antibiotics of 61 microbial keratitis isolates of Pseudomonas aeruginosa from Australia and India using the MBC/MIC ratio, with tolerance defined by a ratio ≥ 32, and tolerance to ciprofloxacin by an agar diffusion assay. MethodsAntibiotics used were ciprofloxacin, levofloxacin, gentamicin, tobramycin, piperacillin, imipenem, ceftazidime and polymyxin B. Isolates were sourced from microbial keratitis infections in Australia and India. Minimum bactericidal and minimum inhibitory concentration (MBC and MIC) were obtained using broth microdilution and compared to breakpoints from the Clinical Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) to determine bacterial susceptibility. Tolerance was assessed as MBC/MIC ≥ 32. An alternative method for tolerance detection (TD) was assessed with 13P. aeruginosa sensitive isolates by agar disk diffusion assay of ciprofloxacin followed by application of glucose to the agar and observation of re-growth of colonies. ResultsThirty-three isolates were resistant to imipenem, 20 to ciprofloxacin, 14 to tobramycin and piperacillin, 12 to levofloxacin and ceftazidime, 8 to gentamicin, and 5 to polymyxin B. The percentage of strains resistant to levofloxacin (7 vs 30 %; p = 0.023), gentamicin (0 vs 24 %; p = 0.005) and tobramycin (4 vs 33 %; p = 0.004) was significantly greater in isolates from India.On average, strains from India exhibited notably greater MIC and MBC values compared to strains obtained from Australia. Out of 61 isolates, none displayed an MBC/MIC ratio ≥ 32. However, three sensitive isolates had low tolerance, nine had medium tolerance and one had high tolerance to ciprofloxacin with the TDtest. ConclusionsThis study used two methods to determine whether P. aeruginosa strains could show tolerance to antibiotics. Using the MBC/MIC criteria no strain was considered tolerant to any of the eight antibiotics used. When 13 strains were tested for tolerance against ciprofloxacin, the most commonly used monotherapy for keratitis, one had high tolerance and nine had medium tolerance. This demonstrates the capacity of P. aeruginosa to develop tolerance which may result in therapeutic failures if inappropriate dosing regimens are used to treat keratitis.

Fosfomycin Resistance in Clinical Isolates of Escherichia coli from Urinary Tract Infections in a Tertiary Care Hospital

Background: Fosfomycin is called an old-new antibiotic, because it was reintroduced for the treatment of urinary tract infections. The rampant use of antibiotics has led to Fosfomycin resistance reported from different parts of the world. The purpose of this study was to find out the frequency of Fosfomycin resistance in clinical isolates of Escherichia coli (E. coli) from urinary tract samples. Methods: This cross-sectional study, including n=314 patients, was conducted in the Microbiology laboratory at Pakistan Naval Ship SHIFA Hospital, Pakistan from January to June 2022. Urine samples were inoculated on agar at 37±2°C for 24-48hrs. The growth of E. coli was confirmed by API 10S as per recommended Clinical Laboratory Standard Institute (CLSI) guidelines. Susceptibility testing was performed by the standard Kirby Bauer Disc Diffusion method. The Chi-square test was applied to categorical variables. p-value ≤ 0.05 was considered statistically significant. Results: Out of 314 clinical isolates, 171 (54.5%) were females and 143 (45.5%) were males (mean age 49±10.3 (6-81) years). Sixty-six (21%) isolates were found resistant to Fosfomycin while 248 (79%) were sensitive. A significant difference was found between the gender (p=0.035), whereas, 29 (17.0%) females and 37 (25.9%) males were found resistant to Fosfomycin. Conclusion: The resistance to Fosfomycin is increasing and it is an impending threat as oral treatment options are limited in urinary tract infections. However, improved surveillance may help in controlling nosocomial infection along with the rational use of antibiotics can prevent and reduce its spread. Keywords: Fosfomycin; Culture; Susceptibility; Urinary Tract Infections (UTIs).

Phenotypic Detection of AmpC \(\beta\)-lactamase Producing Escherichia coli among Patients in Hospital Wards

Background and Objectives: AmpC β-lactamase producing Escherichia coli has emerged in hospital environments as an important health issue that is of a global health concern as drug-resistant pathogenic bacteria that has evolved into strains that are resistant to many classes of antibiotics. However, awareness of the prevalence of AmpC β-lactamase producing microorganisms such as E. coli could be very valuable for achieving more accurate epidemiological results, as well as controlling their spread in the different hospital wards.
 Methodology: A total of four hundred (400) clinical samples comprising of two hundred and seventy-nine (279) urine samples and one hundred and Twenty-one (121) wound samples were collected from patients in different ward at AE-FUTHA. The clinical samples were analyze using standard microbiological culture and identification of Escherichia coli. Detection of phenotypic AmpC β-lactamases production was performed using Cefoxitin-Cloxacillin Double-Disk Synergy Test (CC-DDST). Antibiotic susceptibility studies of AmpC β-lactamases producing Escherichia coli was determined using the Kirby–Bauer disk diffusion method and the results were construed using the Clinical Laboratory Standard Institute (CLSI) zone diameter breakpoints.
 Results: Phenotypic AmpC β-lactamases producing E. coli accounted overall prevalence rate of 94.1%. The frequency of AmpC β-lactamases producing E. coli in urine samples were 57(93.4%) comprising of high occurrence rate in Surgical ward 15(100%) followed by medical ward 24(88.9%) but also accounted for overall prevalence rate of 38(95.0%) in wound samples consisting of 9(100 %) from children ward and medical ward 10(100%) followed by Orthopedic ward 13(86.7%) with the least prevalence rate. The AmpC β-lactamases producing E. coli exhibited high percentage of resistance within the range of 50-100% against ceftriaxone, ceftazidime, cefepime, azetronam, Trimethoprim-Sulfamethoxazole Ticarcillin-clavulanic acid but were susceptible to ofloxacin 70.0%, Imipenem 83.3% and amikacin 100%.
 Conclusion: The findings of this study is a proof of the occurrence of AmpC β-lactamase producing Escherichia coli among patients admitted to hospital wards and there’s an urgent need for controlling and managing the development of MDR genotype strain. Moreover, to prevent the spread of resistance among AmpC β-lactamase producing Escherichia coli to other strains and improve the effectiveness of antibiotics, it is suggested to establish a precise schedule for antibiotic use in each region based on their antibiotic resistance pattern.

Epidemiological cut-off values for non-O1/ non-O139 Vibrio cholerae disc diffusion data generated by standardised methods

This work generates the data needed to set epidemiological cut-off values for disc-diffusion zone measurements of Vibrio cholerae. The susceptibility of 147 European isolates of non-O1/non-O139 V. cholerae to 19 antibiotics was established using a standardised disc diffusion method which specified incubation of Mueller Hinton agar plates at 35°C. Epidemiological cut-off values were calculated by analysis of the zone size data with the statistically based normalised resistance interpretation method. Cut-off values for 17 agents were calculated by analysis of the aggregated data from all 4 laboratories participating in this study. The cut-off values calculated were ≥18 mm for amoxicillin/clavulanate, ≥18 mm for amikacin, ≥19 mm for ampicillin, ≥27 mm for cefepime, ≥31 mm for cefotaxime, ≥24 mm for ceftazidime, ≥24 mm for chloramphenicol, ≥31 mm for ciprofloxacin, ≥16 mm for erythromycin, ≥ 27 mm for florfenicol, ≥16 mm for gentamicin, ≥23 mm for imipenem, ≥25 mm for meropenem, ≥29 mm for nalidixic acid, ≥28 mm for norfloxacin, ≥13 mm for streptomycin and ≥23 mm for tetracycline. For the other 2 agents the data from 1 laboratory was excluded from the censored aggregation because the data from that laboratory was considered excessively imprecise. The cut-off values for these 2 agents calculated for the aggregation of the data from 3 laboratories were ≥23 mm for trimethoprim and ≥24 mm for trimethoprim/sulfamethoxazole. These zone size data will be submitted to the Clinical Laboratory Standards Institute (CLSI) and European Committee for Antimicrobial Susceptibility Testing (EUCAST) for their consideration in setting international consensus epidemiological cut-off values for non O1/non-O139 V. cholerae.

Bacteriological Profile and Antibiotic Susceptibility Pattern in Sterile Body Fluids from a Tertiary Care Hospital in Bhubaneswar, Odisha, India: A Cross-sectional Study

Introduction: Sterile body sites are those where no microorganisms exist as commensals in an otherwise healthy person. Isolation of microorganisms from these sites can indicate either pathological agents or contaminants from the skin. Sterile body fluids are frequently received in microbiology laboratories for culture and sensitivity testing, as the isolation of pathogens from these sites is associated with significant mortality and morbidity. Aim: To assess the current scenario of aerobic bacteriological profiles and their antibiotic susceptibility patterns in various sterile body fluids at a tertiary care hospital in Bhubaneswar, Odisha, India. Materials and Methods: A hospital-based observational study was conducted in the Department of Microbiology at a Tertiary Care Hospital in Bhubaneswar, Odisha, India. The study duration was one year, from January 2022 to December 2022. A total of 450 body fluid samples were collected from 567 patients, of which 117 samples did not meet the inclusion criteria. Out of the 450 samples, 315 were from male patients and 135 were from female patients. All infected body fluids received from clinically diagnosed cases, irrespective of age and gender, were included. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disc diffusion method following the guidelines of the Clinical Laboratory Standards Institute (CLSI). Sterile body fluids were collected with complete aseptic precautions and processed in the Department of Microbiology using standard laboratory procedures. Statistical data analysis was conducted using MS Excel and Statistical Package for Social Sciences (SPSS) version 27.0. A p-value <0.05 was considered statistically significant. Results: A total of 450 clinical samples were processed, of which 153 (34%) were peritoneal fluid, 92 (20.5%) were synovial fluid, 77 (17.1%) were pleural fluids, 118 (26.2%) were Cerebrospinal Fluid (CSF), and 10 (2.2%) were pericardial fluid. In the present study, 126 (28%) pathogens were isolated from the 450 processed samples, with gram negative bacilli being the predominant isolates (88/126, 69.8%), while the remaining 38/126 (30.2%) were gram-positive isolates. Among the 88 gram negative isolates, Escherichia coli was the most common (27, 21.4%), followed by Klebsiella pneumoniae (23, 18.2%), Pseudomonas aeruginosa (14, 11.1%), Acinetobacter spp. (12, 9.5%), Enterobacter spp. (7, 5.6%), and Citrobacter spp. (5, 4.0%). Similarly, among the gram-positive isolates, Staphylococcus aureus was the most common (20, 15.9%), followed by Coagulase-Negative Staphylococci (CONS) (16, 12.7%) and Enterococcus spp. (2, 1.6%). Gram negative isolates showed 100% sensitivity to colistin and polymyxin B, followed by imipenem (90%) and cefepime (80%). Grampositive isolates exhibited 100% sensitivity to linezolid, followed by vancomycin. Conclusion: Early identification of pathogens from these sites, along with their antibiotic susceptibility patterns, will help clinicians initiate targeted therapy. This approach can reduce hospital stays for patients and minimise the development of drug resistance.

Bacterial Superinfection and Antibiotic Management in Patients with COVID-19 Admitted to Intensive Care Medicine in Central Iran: A Follow-Up Study.

Bacterial superinfections are one of the crucial challenges in patients with coronavirus disease 2019 (COVID-19) that are associated with a high mortality rate. The current study was designed to assess bacterial superinfections and antibiotic management in COVID-19 patients admitted to intensive care unit (ICU). Seventy-three adult intubated patients with COVID-19 were included in a cross-sectional study. The lung aspirate samples were collected in two stages and assessed for bacterial growth by standard methods. Antimicrobial susceptibility testing was performed using the Kirby-Bauer method as recommended by the Clinical Laboratory Standard Institute guideline (2021 edition). Also, demographic and clinical data were collected. The statistical analysis was done by chisquare test and Student's t-test, and a P value <0.05 was considered significant. Forty men and thirty-three women with a mean age of 64.78 ± 13.90 have included in our study. The mean length of hospitalization and stay in ICU were 18.77 ± 12.94 and 13.51 ± 9.83 days, respectively; 84.9% of cases died. Thirty-three patients had a bacterial superinfection mainly caused by Klebsiella spp and Acinetobacter spp; 21.2% of piperacillin/tazobactam consumers' patients survived that; the differences were significant (p = 0.034). A significant relationship was seen between superinfection and length of hospital stay until intubation (p = 0.033). Bacterial superinfection and mortality rates were relatively high in COVID-19 patients admitted to ICU. According to the results, using beta-lactam/beta-lactamase inhibitors antibiotics in hospitalized patients in ICU can effectively control superinfection.

Uropathogens and their antimicrobial-resistant pattern among suspected urinary tract infections patients in eastern Nepal: A hospital inpatients-based study.

Urinary tract infections are the primary factors that cause mortality and morbidity in patients with underlying comorbid conditions and are responsible for most hospital admissions worldwide. The study aims to identify the common bacterial uropathogens and determine their antimicrobial susceptibility pattern, including multidrug-resistant/extensively drug-resistant bacteria. The descriptive cross-sectional study was conducted among inpatients provisionally suspected of urinary tract infections in the medical ward of Koshi Hospital, Biratnagar, Nepal. Samples were inoculated in a cystine lysine electrolyte-deficient medium, and pure growth of significant bacteria was further subjected Gram staining, biochemical identification, and antimicrobial susceptibility testing as per laboratory standard procedure and Clinical Laboratory Standards Institute guidelines, respectively. Descriptive and inferential statistical analysis was performed to analyze the outcomes and a p-value < 0.05 was considered statistically significant. A total of 305 patients urine specimens were examined, of which 251 (82.29%) samples resulted in significant bacterial growth in the culture. Escherichia coli (62.94%) was the most predominantly isolated organism, followed by Klebsiella pneumoniae (12.35%), Staphylococcus aureus (9.16%), and Pseudomonas aeruginosa (8.76%). Among antimicrobials, colistin had shown absolute susceptibility (100%) toward gram-negative uropathogens followed by carbapenem and aminoglycosides in a majority of uropathogens. Escherichia coli was found to be the leading drug-resistant bacteria (70%) among uropathogens. The presence of multidrug-resistant/extensively drug-resistant bacteria uropathogens was found to be significantly associated with diabetes mellitus and those with combined antimicrobial therapies. Diabetic patients were twice (OR~2) more likely to colonize and develop uropathogens as compared to non-diabetics. Escherichia coli was the most common uropathogens followed by Klebsiella pneumoniae in urinary tract infection patients. The polymyxin group (colistin) of antimicrobials was found to be effective in all multidrug-resistant and extensively drug-resistant uropathogens. The study recommends the need of optimized antimicrobial stewardship program to develop effective strategies in the management of urinary tract infections in diverse healthcare settings.

Antimicrobial susceptibility pattern of extended-spectrum beta-lactamase-producing uropathogens in aminu Kano teaching hospital, Northwestern Nigeria.

UTI is the most common nosocomial infection among hospitalized patients. Emerging increasing resistance has been observed among uropathogens of the family enterobacterales. Extended spectrum beta lactamase genes encode for multidrug resistance and unrestricted use of antibiotics in hospitals provides an environment for spread of infections with limited treatment options. This necessitates therapies based on culture and antimicrobial sensitivity to improve patients' outcomes We aimed to determine susceptibility pattern of ESBL uropathogens among hospitalized patients in Aminu Kano Teaching Hospital, Kano. Three hundred and eighty-nine urine samples were obtained from in-patients with UTI between April 2020 and April 2021. Five samples were rejected and remaining analyzed. Susceptibility testing was done by modified Kirby Bauer technique. Clinical Laboratory Standards Institute guidelines 2019 (CLSI 2019) and the European Committee on Antimicrobial Susceptibility testing guidelines version 2 (EUCAST version 2) were used for screening and confirmation of ESBL production respectively. Of the 384-urine processed, 105 (27.3%) were gram negatives and 81 were Enterobacterales, Isolation rates were E. coli-55.6%, K. pneumoniae-29.6%, Citrobacter spp.-12.3%, P. mirabilis-1.2% and Morganella spp.-1.2%. Among Enterobacterales, 32 (39.5%) were ESBL producers. Prevalence of ESBL were 62.5% for Escherichia coli, 28.1% for Klebsiella pneumoniae and 9.4% for Citrobacter species. Susceptibility showed that ESBL-producing Isolates were highly susceptible to amikacin (96.9%). Resistance to other antibiotics varied from 3.1% to 100%. We recommend strengthening laboratory capacity, antibiotics stewardship and Infection control to prevent spread of resistant pathogens including ESBLs.

Characterization of Extended Spectrum β-lactamase and AmpC Producing Organisms Isolated from a Third Level Hospital Patients in Dominican Republic

Extended-spectrum β-lactamases (ESBLs) producing Enterobacteriaceae have become a challenge for clinicians as they frequently show co-resistance to other antibiotic classes. AmpC-type β-lactamases (AmpCs), tend to be more difficult to treat as they could be induced during antibiotic therapy. To characterize ESBLs and AmpCs producing organisms, we collected clinical samples identified through MicroScan® as members of the Enterobacteriaceae family and ESBLs producers, following Clinical Laboratory Standards Institute indications, from January through March 2021, in a third level hospital in Santiago de Los Caballeros, Dominican Republic. Samples underwent manual confirmation via ESBL + AmpC screen discs kit (Liofichelm® srl, Italy) and a genotype analysis was done by DNA extraction for detection of specific genes related with ESBL ((blaSHV, blaTEM, blaCTX, and blaCMY-4) or AmpC (CIT, MOX, DHA and FOX) expression. 54 samples were confirmed as ESBL and/or AmpC producers with Escherichia coli as the most frequent (33/61%) followed by Klebsiella sp. (9/17%). blaCTX was detected in 29 (67%) isolates, followed by blaTEM (23/53%) and blaSHV (18 /40%) . Among the AmpC encoding genes, DHA and CIT gene pools were detected in 3 (7%) and 2 (5%) of the samples, respectively. Two or more genes were detected in 19 (44%) samples. All the AmpC genes were found in an isolate which already had, at least, two other β-lactamase encoding genes. In this cohort, ESBLs and AmpCs -producing organisms with multiple resistance genes were detected. Efforts to curb antibiotic availability and to establish antimicrobial stewardship programs are needed.

Comparison of the Two Coagulation Techniques: Thrombolyzer XRM and Diagon Coag XL.

The aim of this study was to evaluate the performance characteristics of the Thrombolyzer XRM against the Diagon Coag XL during the exchange of the coagulation analyzer. The Partial Thromboplastin Time (PTT) and prothrombin time (PT) measurements were performed by using the Thrombolyzer XRM and Diagon CoagXL analyzers. The precision, accuracy, method comparison, and reference range verification studies were performed on the Thrombolyzer XRM based on Clinical Laboratory Standards Institute guidelines. The precision study was performed with normal and pathologic controls for the PT and PTT tests, all co-efficients of variation (%) were found ≤ 2.5%. In the accuracy study, both analyzers displayed less than 6.3% and 10.8% bias for PT and PTT tests, respectively. The method comparison study demonstrated good agreement for the Bland-Altman plots for PTT and PT with a bias of -2.0 and -3.3 between each analyzer, respectively. The Passing and Bablok analysis showed no significant differences for PTT and PT between each analyzer (p = 0.65 and p = 0.33, respectively). However, there was a proportional bias for PT with a slope of 1.40 (95% CI 1.2 - 1.8). The manufacturer ranges were acceptable as a result of the reference range verification study. The routine coagulation analysis can be performed on the Thrombolyzer XRM with satisfactory precision and the obtained minor differences can be eliminated in future standardization studies.

Luliconazole: A Novel Potent Imidazole Activity against Aspergillus niger and Aspergillus flavus Causing Otomycosis

Background: Aspergillus niger and Aspergillus flavus are the most common causative agents of otomycosis with worldwide distribution. Extensive surgical debridement and systemic antifungal therapy are needed in cases of refractory otomycosis. Luliconazole is currently confirmed for the topical therapy of dermatophytosis. Moreover, it is found that luliconazole has in vitro activity against some molds and yeast species. Objectives: The aim of the present study was to evaluate the efficacy of luliconazole in comparison to routinely used triazole antifungals on Aspergillus isolates, obtained from otomycosis cases using microbroth dilution method. Materials and Methods: This study was carried out in the department of microbiology, from March 2020 to February 2022. A total of 55 strains of Aspergillus isolates obtained from otomycosis cases were confirmed based on macroscopic and microscopic identification. Antifungal susceptibility patterns of itraconazole, voriconazole, posaconazole and luliconazole were determined by broth microdilution method as per the Clinical Laboratory Standards Institute M38-A2 guidelines. Results: Among the azoles tested, the lowest minimum inhibitory concentration (MIC) geometric mean (0.00309 μg/ml), MIC50 (0.00098 μg/ml), and MIC90 (0.00781 μg/ml) values were attributed to luliconazole. Being the drug of choice for aspergillosis, voriconazole had a slightly higher MIC50 and MIC90 value of 1 μg/ml and 2 μg/ml, respectively. Even for the pan azole resistant isolates (n = 3), luliconazole was found to be more effective with lower MIC values. Conclusion: The results of this study showed that luliconazole had an excellent in vitro activity with a lower MIC values than the triazoles tested. Hence, this novel imidazole antifungal agent can be considered an appropriate candidate for the treatment of otomycosis caused by A. niger and A. flavus strains. Furthermore, luliconazole showed better efficacy with lower MIC values for pan azole resistant isolates, suggesting that it could be a potential antifungal for treating aspergillosis caused by pan azole resistant isolates.

Non-parametric Clinical Laboratory Reference Interval Estimation in Volunteer Blood Donors: An Example for Prothrombin Time and Partial Thromboplastin Time

Introduction. To date, there are no reference intervals for prothrombin time (PT) and activated partial thromboplastin time (APTT) based on “normal” Filipino adults. The common practice in most laboratories is to adopt manufacturer provided values or foreign literature even if the importance of establishing or at least verifying laboratory reference intervals has been stressed by Clinical Laboratory Standards Institute (CLSI). Objectives. Here we aim to describe our experience in using a simple non-parametric method to generate reference intervals for PT and APTT, from healthy Filipino volunteer blood donors. Methodology. We used a de novo, a priori non-parametric estimation method following the CLSI guidelines on establishing reference intervals. Results. The non-parametric lower reference limit for PT is 12.55 seconds, with 90% confidence interval of 12.3 to 12.75 seconds. While the non-parametric upper reference limit for PT is 16.15 seconds, with 90% confidence interval of 15.55 to 16.55 seconds. The non-parametric lower reference limit for activated partial thromboplastin time is 26.12 seconds, with 90% confidence interval of 22.95 to 27.1 seconds, and the non-parametric upper reference limit for activated partial thromboplastin time is 37.44 seconds, with 90% confidence interval of 36.75 to 38.65 seconds. The PT and APTT reference intervals were different from foreign sources and manufacturer provided values in terms of interval width and values of the reference limits by 2 to 4 seconds. Conclusion and Recommendations. Estimation of coagulation reference intervals from volunteer health blood donors is doable, simple, and practical. Collaborative multi-center efforts may be done to expand the pool of reference individuals that are included and increase the representativeness of the reference intervals generated. This simple method can also be used to generate reference intervals for other clinical laboratory assays and may also be extended to at least verify reference intervals in special populations like pregnant women, the elderly, and the pediatric population.

Antibiotic resistance pattern of Methicillin-resistant Staphylococcus aureus in asymptomatic prisoners of Kathmandu valley

bjectives: The purpose of this study was to determine the prevalence of methicillin-resistant Staphylococcus aureus and its antimicrobial resistance among the prisoners within the prisons of Kathmandu valley. Methods: A total of 320 nasal samples were collected from the prisoners. S. aureus was confirmed by using standard microbiological techniques including culture and biochemical tests. MRSA was confirmed using cefoxitin disc and antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion method results were interpreted according to the guidelines of Clinical Laboratory Standard Institute (CLSI). Results: Out of 320 samples, 41 (12.82%) isolates were confirmed S. aureus and 8 (19.51%) of them were screened as MRSA positive. Highest percentage of MRSA was found in the samples collected from Nakhu prison 6 (75%) followed by Central prison 1 (12.5%) and Juvenile Correctional Home 1 (12.5%). Majority of the of MRSA isolates (62.5%) were also found to be multiple drug-resistant strains. All of the MRSA (100%) were sensitive towards linezolid and resistant towards ciprofloxacin. Conclusion: The presence of methicillin-resistant S. aureus among the prisoners indicates the emergence of drug resistance in a vulnerable population which may lead to hazardous disease breakouts in the prison and even in the community after their release. Regular monitoring and its minimization should be done in order to control its dissemination.

Methicillin-Resistant Staphyloccocus aureus Contamination in Hospital Door Handles of Pokhara, Nepal

The presence of contaminants on the door handles surfaces of hospitals may be a common means of transfer of potentially pathogenic bacteria among users and can act as vehicles of diseases transmission. The aim of this study was to isolate, identify and evaluate the presence of bacterial contaminants and Methicillin Resistant Staphylococci (MRSA) with their antimicrobial susceptibility pattern present on the door handles of selected hospitals of Pokhara Metropolitan City, Western Nepal to take the necessary remedial measures. Isolation, identification and antimicrobial susceptibility test of bacteria were done using standard microbiological procedures. Further screening and confirmation of MRSA were done according to Clinical Laboratory Standard Institutes (CLSI) guidelines. Out of the 100 swab samples cultured, 96 (96%) showed bacterial growth. 62.86% (88/140) of the isolates were found to be Gram positive. Percentage distribution of the isolates showed that the most prevalent Gram-positive bacteria identified was Staphylococcus aureus (43.18%), followed by Coagulase negative Staphylococci (15.91%), Bacillus spp (11.36%), Diptheroides (10.23%) but considerable number of Micrococci (7.95%), Streptococcus pneumoniae (6.82%), Enterococci (4.55%). Prevalence of MRSA in this study was 39.47%. Significant difference in antibiotic resistance pattern was found among MRSA and MSSA isolates (P&lt;0.05) reflecting increased ability of MRSA to develop resistance against various antimicrobials. Drugs like clindamycin, cefazoline and amikacin were found quite effective against MRSA in the present study would be better options for the management of such infections. MRSA strains’ contaminating on the door handles of hospitals may cause threat of infections to patients, hospital staffs, visitors to hospitals and people in the community. Thus, regular surveillance and disinfection with appropriate agent at regular interval would minimize the settlement and transmission of various pathogens including MRSA.Keywords: Contaminants; disinfection; door handles; hospital; MRSA; pathogen

Prevalence and Antibiogram Profile of Carbapenem-resistant Escherichia coli and Klebsiella pneumoniae among Patients with Urinary Tract Infection in Abakaliki, Nigeria

Background and Objectives: Carbapenem antibiotic are drug of last-resort from the treatment of bacterial infection, as a result of the prevalence and rapidly evolving enzymes from Carbapenem resistant bacteria such Escherichia coli and Klebsiella pneumoniae make urinary tract infection difficult, and in some cases impossible to treat in health care settings. With limited progress of new antibacterial drugs, the best approach is monitoring the prevalence and antibiogram profile of carbapenem-resistant Escherichia coli and Klebsiella pneumoniae among patients with UTI in Abakaliki, Nigeria.&#x0D; Methodology: A non-repetitive, clean catch mid-stream urine was collected from five hundred (500) diagnosed UTI inpatient and outpatient. The samples were evaluated using routine microbiological protocol for isolation and identification of Escherichia coli and Klebsiella pneumoniae. Phenotypic screening of Carbapenem-resistant strains was performed using Modified Hodge Testing. Antibiogram studies of carbapenem-resistant Escherichia coli and Klebsiella pneumoniae was performed using the Kirby–Bauer disk diffusion method and the results were interpreted using the Clinical Laboratory Standard Institute (CLSI) zone diameter breakpoints. Multiple antibiotic resistance index (MARI) was determined for MDR strain.&#x0D; Result: The prevalence of Escherichia coli and Klebsiella pneumoniae isolate accounted for 148(29.6 %) consisting of 95(54.3 %) and 53(16.3 %) from in-patients and out-patients. Escherichia coli accounted overall isolation rate of 112(22.4 %) comprising of high proportion among in-patient 82(46.9 %) over out-patient 30(9.2 %). The proportion of K. pneumoniae accounted for 36(7.2 %) with 13(7.4 %) and 23(7.1 %) recorded among in-patients and out-patients. Association between presence of Escherichia coli and Klebsiella pneumoniae isolates in clinical samples was statistically significant with patient’s population with p value &lt;0.05. Carbapenem-resistant Escherichia coli and Klebsiella pneumoniae accounted for 37(7.4 %) comprising of 24(13.7) and 13(4.0 %) among in-patients and out-patients respectively while carbapenem-susceptible Escherichia coli and Klebsiella pneumoniae accounted for overall detection rate of 111(22.2 %) consisting of 71(40.6 %) and 40(12.3 %) among in-patients and out-patients respectively. The isolates resistance rate to cephalosporins were relatively high i.e., Cefotaxime, Cefoxtin Ceftazidime, Ceftriaxone resistance was observed at 60-100% while amoxicillin/clavulanate, azetronam, tetracycline nitrofurantoin and Ticarcillin-clavulanic acid recorded 100 % with MDR index ranged from 0.5-0.8, but were 100 % and 85.0 % sensitive to ciprofloxacin and ofloxacin.&#x0D; Conclusion: These results strongly hypothesize that MDR bacteria, including Carbapenem-resistant isolate, have become common residents in various hospital environments, however with substantial evidence in this study, ciprofloxacin and ofloxacin as drugs of choice could be used for treatment of UTI. Therefore, its importance that good antibiogram evaluation of other drug classes beside fluoroquinoles reported in this study need to be establishes as baseline for empirical diagnosis, epidemiological surveillance, drug prescriptions and infection management.

Antibiotic Susceptibility Profile of Clinical Isolate of Carbapenem-Resistant Pseudomonas aeruginosa

Background and Objectives: Carbapenem-resistant Pseudomonas aeruginosa, are among the top tier of the list of antibiotic-resistant priority pathogens that pose the greatest threat to human health. In recent years, the rate of carbapenem resistance in Pseudomonas aeruginosa has increased worldwide and has become of great concern since it significantly restricts the therapeutic options for patients. Therefore this study was undertaken to determine the antibiotic susceptibility profile of the clinical isolate of Carbapenem-resistant Pseudomonas aeruginosa.&#x0D; Methodology: A total of five hundred (500) clinical samples were collected from patient’s attending Alex Ekwueme Federal University Teaching Hospital Abakaliki, Ebonyi State (AFEUTHA). The collected samples were analyzed for the presence of Pseudomonas aeruginosa using standard microbiological techniques for isolation and characterization of bacteria. Further strain confirmation was performed using VITEK 2 System. Phenotypic detection of Carbapenem-resistant Pseudomonas aeruginosa was performed using Modified Hodge testing. Antibiotic susceptibility was performed by employing Kirby-Bauer disk diffusion method and the results were interpreted using the Clinical Laboratory Standard Institute (CLSI) zone diameter breakpoints.&#x0D; Results: The occurrence rate of Pseudomonas aeruginosa in clinical samples accounted for 119(23.8%) consisting of a high proportion from urine sample 81(27.4%) followed by wound swabs 13(25.5%), high vaginal swabs 17(20.7) while the least occurrence rate was observed against catheter tips 5(12.8%) and sputum 3(9.4%). Modified Hodge testing revealed 31(6.2%) carbapenem-resistant Pseudomonas aeruginosa comprising of high proportion of 24(8.1%) from urine samples followed by wound swab 5(9.8%) while Carbapenem-resistant Pseudomonas aeruginosa was absent in High Vaginal Swab recording 0(0.0%). Carbapenem-resistant Pseudomonas aeruginosa isolates were highly resistant to amoxicillin-clavulanic 100%, colistin 100%, tetracycline 100%, nitrofurantoin 70.8%, aztreonam 87.5% but were susceptible to nalixidic acid 50.0 %, ofloxacin 75.0%, and ciprofloxacin 100%.&#x0D; Conclusion: As in-vitro susceptibility of carbapenem-resistant Pseudomonas aeruginosa isolates to ofloxacin and ciprofloxacin is known, their judicious utilization will accelerate a significant improvement in the patient's condition. As such, there is a substantial need for the evaluation of a wide spectrum and new therapies in different classes to counteract this imminent crisis of resistance among Carbapenem-resistant Pseudomonas aeruginosa.