ABSTRACT
Urinary tract infections (UTIs) are highly prevalent globally, and various antibiotics are employed for their treatment. However, the emergence of drug-resistant uropathogens towards these antibiotics causes a high rate of morbidity and mortality. This study was conducted at the Microbiology Laboratory of Grande International Hospital from November 2021 to May 2022 and aimed to assess the prevalence of UTI caused by Escherichia coli and their antibiotic susceptibility pattern with a focus on extended-spectrum beta-lactamases (ESBLs) and the prevalence of two genes (blaCTX-M and blaTEM) in cephalosporin-resistant E. coli. Altogether, 1050 urine samples were processed to obtain 165 isolates of E. coli. The isolates were identified by colony morphology and biochemical characteristics. Antimicrobial susceptibility tests (ASTs) were determined by the Kirby-Bauer disk diffusion method, and their ESBL enzymes were estimated by the combined disk method (CDM). Two ESBL genes (blaCTX-M and blaTEM) were investigated by polymerase chain reaction (PCR) in cefotaxime-resistant E. coli. Among the 1050 urine samples that were processed, 335 (31.9%) were culture-positive with 165 (49.2%) identified as E. coli. The age group ≥60 years (30.3%) had greater susceptibility to bacterial infections. AST revealed that meropenem was highly effective (95.7% susceptibility), while ampicillin showed the least sensitivity (42.4%). Among the E. coli isolates, 86 were multidrug resistant (MDR) and 10 were extensively drug resistant (XDR). Of these, 46 MDR (96%) and 2 XDR (4%) were ESBL producers. The prevalence of ESBL genes (blaCTX-M and blaTEM) was 49.3% and 54.8%, respectively. The overall accuracy of CDM as compared to PCR for the detection of the blaCTX-M gene was 55.26%. The prevalence of MDR E. coli harboring the blaCTX-M and blaTEM genes underscores the imperative role of ESBL testing in accurately identifying both beta-lactamase producers and nonproducers.
ABSTRACT
Background: 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. Objectives: The study aims to identify the common bacterial uropathogens and determine their antimicrobial susceptibility pattern, including multidrug-resistant/extensively drug-resistant bacteria. Methods: 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. Results: 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. Conclusion: 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.
ABSTRACT
Chryseobacterium indologenes is gram-negative bacteria that cause infection in humans. It is less frequently isolated in the laboratory. The development of drug-resistant and its intrinsic ability to resist a wide range of antimicrobials enables them to cause mortality in an immunocompromised patient with a longer hospital stay. Our study objectives are to investigate antimicrobial-resistant patterns, drug-resistant enzymes, and the risk factor analysis associated with multidrug-resistant (MDR), extensively drug-resistant (XDR), and Pan-drug resistant (PDR) within 2 years. Altogether 53 strains of Chryseobacterium indologens were obtained from 5000 specimens that were processed for routine bacterial culture. The bacterial identification was done using conventional techniques (colony morphology, gram staining, flexirubin test, and biochemical tests) as well as the VITEK-2 System to further confirm. The bacterial isolate were processed to observe antimicrobial susceptibility test (AST) using disk diffusion method. MDR XDR and PDR were classified following European Centre for Disease Prevention and Control guidelines. C. indologens strains with beta-lactamases such as extended-spectrum beta-lactamases (ESBL), metallo beta-lactamases (MBL), and Amp-C beta-lactamases (Amp-C) were detected phenotypically. The highest isolation of C. indologens was observed in a sputum sample. In vitro antimicrobial susceptibility test revealed susceptibility to tigecycline followed by levofloxacin, cotrimoxazole, and piperacillin-tazobactam. From 53 isolates of C. indologens, MDR accounts for 56.60% and 22.64% for XDR. Combined antimicrobial therapy and longer hospital stay were found to be the leading risk factor. All 53 C. indologenes strains were detected as MBL. Total ESBL was detected in 16.98% of MBL producer strains and Amp-C was observed in 13.20% of MBL-producing strains. All 3 enzyme co-oproducers were seen in only 5.66% of C. indologens. Although it is rarely encountered in the laboratory, it showed a remarkable effect in patients with underlying predisposing factors and prolonged hospital stays. The presence of betalactamases determined the drug-resistant activity on a wide spectrum of tested antibiotics.
ABSTRACT
Fungal infections of hair, nail, and skin are common worldwide and tend to increase. The present study was conducted to determine the prevalence of dermatomycoses, estimate the efficiency of rapid potassium hydroxide (KOH) wet-mount, and observe the hygienic status and the predisposing risk factors. Altogether 115 samples (nail = 77, skin = 30, and hair = 8) were obtained in a duration of December 2019 to June 2020 at Grande International Hospital, Nepal. The samples were examined by KOH wet-mount microscopy and further processed for culture. The dermatophyte test medium (DTM) was used to isolate dermatophytes separately. The fungal colonies obtained in SDA, SDA with cycloheximide/chloramphenicol and dermatophyte medium were subjected to lactophenol cotton blue (LPCB) reagent to study fungal morphology. The yeast colonies grown on SDA were subjected to Gram staining, germ-tube tests, and biochemical tests for identification. CHROMagar was used to distinguish different Candida species based on its pigment production in the medium. Various factors (age, sex, occupation, and hygiene condition) were analyzed which were associated with mycological infection. Out of 115 samples, the presence of fungal elements was detected in 20 samples by KOH. Nondermatophyte molds were the most isolated fungus in nails, skin, and hair, followed by yeast and dermatophytes, respectively. Dermatomycosis molds were the most common causative agents with 22 (14.7%) cases, followed by yeasts with 6 (5.21%) cases. Candida albicans was isolated from 5 (4.3%) cases, whereas Rhodotorula species accounted for a single (0.8%) case. Dermatophytes were isolated from 5 (4.3%) cases. Among them, n = 4(3.4%) cases revealed Trichophyton rubrum and Trichophyton mentagrophytes was isolated from single (0.8%) case. The most isolated nondermatophyte mold that follows criteria as a pathogen in our study was Cladosporium species 6 (25%) out of 27 total fungal isolates. Poor hygiene and sweating were found to be statistically significant (P < 0.05) in fungal cases detected by both KOH and culture. Dermatophytes and nondermatophyte fungi were emerging as important causes of fungal infection. Both direct microscopy and culture followed by LPCB together were vital tools for the diagnosis of fungal infections.
ABSTRACT
For ongoing malaria elimination programmes, available methods such as microscopy and rapid diagnostic tests (RDTs) cannot detect all malaria cases in acute febrile illness. These methods are entirely dependent on the course of infection, parasite load, and skilled technical resources. Our study objectives were to estimate the performance of light microscopy and a RDT as well as real-time PCR for the detection of the Plasmodium parasite. Altogether, 52 blood samples collected from patients with acute febrile illness were tested by microscopy, RDT, and real-time PCR. The results were compared in terms of sensitivity and specificity. Microscopy detected the malaria parasite in 5.8% of the blood samples whereas 13.5% were detected by the RDT and 27% by real-time PCR. Considering real-time PCR as the gold standard method, microscopy had a sensitivity of 21.4% and a specificity of 100%, and the RDT had a sensitivity of 28.6% and a specificity of 92.1%. Microscopy together with the RDT successfully detected malaria positive cases in blood samples of Ct value below 20, but both were unable to detect malaria cases between 26-40 Ct value ranges amplified by real-time PCR. Despite various diagnostic tools being available, microscopy still remains the method of choice for diagnosis, while the RDT is user-friendly when applied at the point of care. However, our preliminary results emphasize the need to implement the test with higher sensitivity and specificity in the context of a malaria elimination programme. Such programmes can be a crucial opportunity to understand the species prevalent in a low-endemic region. However, these results should be further verified with a large cohort study to document the submicroscopic infection.
