Phenotypic characterization of ESBL-producing urinary isolates of E. coli and Klebsiella spp. in a tertiary care children's hospital in Nepal.

BACKGROUND: The production of extended-spectrum beta-lactamases (ESBLs) among uropathogens, particularly E. coli and Klebsiella spp., poses a severe public health concern. This study explored the epidemiology of ESBL-producing E. coli and Klebsiella spp. isolated from urine samples obtained at a tertiary care children's hospital in Nepal. METHODS: A cross-sectional study was conducted from August 2016 to February 2017. A total of 745 clean catch urine samples were obtained from pediatric patients under the age of 13 and subjected to semiquantitative culture. E. coli and Klebsiella spp. were identified using standard laboratory protocols. Antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method, and ESBL producers were phenotypically identified using the combined disk method. RESULTS: Among the bacterial isolates, E. coli predominated, accounting for 139 (81.8%) positive cases. Notably, E. coli showed high susceptibility to nitrofurantoin, with 117 (84.2%) isolates being susceptible. Meanwhile, K. pneumoniae showed high susceptibility to gentamicin, with 21 (91.3%) isolates being susceptible. Of the 163 isolates of E. coli and Klebsiella spp., 62 (38.0%) were identified as multidrug-resistant (MDR), with 42 (25.8%) confirmed as phenotypic ESBL producers. Remarkably, all 41 (100%) ESBL-producing E. coli isolates were susceptible to imipenem. CONCLUSIONS: The prevalence of ESBL producers among E. coli and K. pneumoniae isolates from pediatric patients underscores the importance of antimicrobial stewardship. Nitrofurantoin and gentamicin emerge as effective empirical treatment choices against these pathogens in children. However, the high rates of multidrug resistance and ESBL production highlight the necessity for routine surveillance, and early detection strategies to manage such infections effectively.

Diagnostic performance of GeneXpert MTB/RIF in detecting MTB in smear-negative presumptive TB patients.

BACKGROUND: Tuberculosis (TB) is a leading cause of morbidity and mortality worldwide. Control of TB is lingering by the lack of diagnostic tests that are simple, rapid, yet accurate. Thus, smear-negative pulmonary TB often misses the diagnosis. The study evaluated the performance of GeneXpert MTB/RIF assay for the detection of Mycobacterium tuberculosis (MTB). METHODS: The study was carried out from June to December 2016 in Nepal Tuberculosis Center, Bhaktapur, Nepal. A total of 173 sputum samples were collected and processed by microscopy [Auramine-O staining and Ziehl-Neelsen (ZN) staining], followed by GeneXpert MTB/RIF assay and culture in Lowenstein-Jensen (LJ) medium. RESULTS: Of 173 sputum samples, 162 (93.6%) were smear-negative. Of 162 smear-negative sputum samples, 35 (21.6%) were confirmed to have MTB by culture, and 31 (19.1%) by GeneXpert MTB/RIF assay. Of 31 GeneXpert-positive samples, 25 (80.6%) were susceptible, 4 (12.9%) were resistant, and 2 (6.45%) were intermediate to rifampicin. The sensitivity, specificity, positive predictive value, and negative predictive value of GeneXpert MTB/RIF assay for smear-negative sputum samples were 74.3%, 96.6%, 86.7%, and 92%, respectively. The GeneXpert MTB/RIF has a substantial diagnostic agreement of 90.91% with culture (Cohen's Kappa coefficient = 0.73). CONCLUSION: The diagnostic performance of GeneXpert MTB/RIF assay was almost on par with culture, and thus can be relied upon for MTB detection in smear-negative sputum samples.

Staphylococcus aureus with inducible clindamycin resistance and methicillin resistance in a tertiary hospital in Nepal.

BACKGROUND: Staphylococcus aureus is a global public health issue in both community and hospital settings. Management of methicillin-resistant S. aureus (MRSA) infections are tough owing to its resistance to many antibiotics. Macrolide-lincosamide-streptogramin B (MLSB) antibiotics are commonly used for the management of MRSA. This study was aimed to determine the occurrence of inducible clindamycin- and methicillin-resistant S. aureus at a tertiary care hospital in Kathmandu, Nepal. METHODS: A total of 1027 clinical samples were processed following standard laboratory procedures and antibiotic susceptibility testing of S. aureus was performed by disc diffusion method. MRSA isolates were detected phenotypically using cefoxitin disc, and inducible clindamycin resistance was detected phenotypically using the D-zone test. RESULTS: Of 1027 samples, 321 (31.2%) were culture positive, of which 38 (11.8%) were S. aureus. All S. aureus isolates were susceptible to vancomycin, and 25 (67%) of S. aureus isolates were multidrug-resistant. Similarly, 15 (39.5%) of S. aureus were MRSA and 14 (36.5%) were inducible clindamycin-resistant phenotypes. CONCLUSION: Inducible clindamycin and methicillin resistance were common in S. aureus. This emphasizes that the methicillin resistance test and the D-zone test should be incorporated into the routine antibiotic susceptibility testing in hospital settings.

Intestinal Parasitic Infections among Prison Inmates in Kathmandu Nepal.

BACKGROUND: Prison inmates are at high risk of intestinal parasitic infections. Thus, we studied intestinal parasitic infections among inmates of the Central Jail, Kathmandu, Nepal. METHODS: Morning stool samples from 400 inmates (M=282 and F=118), were collected in a clean, dry and wide-mouthed plastic container. The samples were transported to the research laboratory of Shi-Gan International College of Science and Technology, and were fixed using 10% formal saline. Then, samples were processed by formal ether sedimentation concentration technique and were observed microscopically by direct-smear technique. RESULTS: Six percent (24/400) samples were positive for intestinal parasites, with a gender ratio (M:F) 1.7:1. But, co-parasitism was not observed. Intestinal parasitic infections were higher among 21-40 years age-group, 3.5% (14/262). Similarly, intestinal parasitic infections were higher among 'Dalits' ethnic group, 21.1% (4/19). As compared to helminths, more protozoans, 62.5% (15/24), were observed. Giardia lamblia, 41.67% (10/24), was the most common protozoans while Trichuris trichiura, 25.0% (6/24), was the most common helminths. CONCLUSIONS: Intestinal parasitic infections were lower among the inmates of Central jail, but such conditions cannot be presumed in other peripheral settings. Pure and safe drinking water supply and the effective deworming campaign can further reduce this figure at this setting and peripheral jails across the country.

Epidemiology of urinary tract infection and antimicrobial resistance in a pediatric hospital in Nepal.

BACKGROUND: Urinary tract infection is an infection affecting infants and children. The aim of this study was to determine the etiology of urinary tract infection along with their antimicrobial resistance. METHODS: This cross-sectional study was conducted from June 2015 to January 2016 at Siddhi Memorial Hospital, Bhaktapur, Nepal. Urine samples were first cultured on cystine lactose electrolyte deficient agar and blood agar by semi-quantitative technique, and then incubated aerobically for 18-24 h at 37 °C. The identified bacterial isolates were tested for antimicrobial susceptibility by Kirby Bauer disc diffusion technique. RESULTS: Of 1599 urine samples, 12.3% samples showed significant bacterial growth. E. coli (58.7%) was the most common pathogen, followed by Klebsiella pneumoniae (22.5%). Most of the isolates were resistant to ampicillin and co-trimoxazole, while least were resistant to amikacin and nitrofurantoin. Higher multi-drug resistance (61.9%) was observed among isolates. CONCLUSIONS: E. coli and Klebsiella spp. were predominant cause of pediatric urinary tract infection in children. Higher susceptibility observed against aminoglycosides and nitrofurans make these drugs suitable in emergency.

Biofilm Production and Antimicrobial Resistance among Uropathogens in Pediatric Cases: a Hospital Based Study.

BACKGROUND: The study was designed to provide account of etiological agents of urinary tract infection in pediatric patients and the antimicrobial resistance pattern plus biofilm producing profile of the isolates. METHODS: The prospective study was conducted in Alka Hospital, Nepal with 353 clean catch urine samples from children. It was obtained during July 2014 to January 2015 which were first cultured by semi-quantitative method, followed by antimicrobial susceptibility testing and biofilm production assay on Congo red agar. Multidrug- resistance, extensively drug- resistance and pandrug- resistance among isolates were considered as per international consensus. RESULTS: Out of 353 samples, 64 (18.13%) showed positive growth in culture, confirming urinary tract infection. E. coli, 44 (68.8%) was the predominant organism followed by Klebsiella spp. 6 (14.1%). Most E. coli were sensitive to amikacin (93.2%) followed by nitrofurantoin (86.4%), and highly resistant to ampicillin (95.5%). Of 64 isolates, 23 (35.93%) were found to be multidrug- resistant strains. Biofilm was produced by 36 (56.25%) isolates. CONCLUSIONS: This study showed higher biofilm production and resistance to in-use antibiotics rendering ineffective for empirical use. Regular surveillance of resistance patterns should be done to regulate multidrug- resistant bugs and to ensure effective management of urinary tract infection in children in a tertiary care setups.

A hospital based surveillance of metallo-beta-lactamase producing gram negative bacteria in Nepal by imipenem-EDTA disk method.

BACKGROUND: A rising threat of the rapid spread of acquired metallo-beta-lactamases (MBLs) among major Gram-negative pathogens is a matter of public health concern worldwide. Hence, for a low income nation like Nepal, surveillance data on MBL producing clinical isolates via a cost effective technique is necessary to prevent their dissemination as well as formulation and regulation of antimicrobial stewardship policy. METHODS: The prospective study was conducted at Nepal Medical College, Kathmandu from May to October, 2014 to assess the prevalence of MBL production among ceftazidime-resistant Gram-negative rods (GNRs) isolates. The samples were processed according to standard microbiological procedure following the Manual of clinical Microbiology. Isolated GNRs were subjected to susceptibility testing against the selected panel of antibiotics by Kirby- Bauer disc diffusion method and interpretation made in conformity with the Clinical and Laboratory Standards Institute (CLSI) guidelines. Ceftazidime-resistant isolates were subjected to the detection of MBL production by imipenem-EDTA combined disc (CD) method. RESULTS: Among the Gram-negative isolates, 5.80% (21/362) were found to be MBL positive with Acinetobacter spp. showing the highest prevalence i.e. 85.71% (18/21), followed by P. aeruginosa i.e. 14.29% (3/21). None of the other cefazidime resistant gram negative bacteria tested were found to be positive for MBL production with all the positive isolates determined to be Multidrug resistant (MDR) strains. CONCLUSION: This study demonstrated a higher rate of resistance among P. aeruginosa and Acinetobacter spp. to a wide variety of antibiotic categories with an additional burden of MBL production within them, warranting a need for strict surveillance and rapid detection of MBL production among the GNRs.

ESBL Production Among E. coli and Klebsiella spp. Causing Urinary Tract Infection: A Hospital Based Study.

INTRODUCTION: Increase in extended-spectrum ß-lactamases (ESBL) producing microbes in recent years has led to limitations of treatment options. This study aimed to assess the prevalence of ESBL producing E. coli and Klebsiella spp. at a tertiary hospital in Nepal. METHODS: A total of 2209 non-repetitive mid-stream urine (MSU) samples were collected during the study period (March to September 2014). Identification of the isolates was done by Gram's staining followed by biochemical tests. Antibiotic susceptibility testing was done by modified Kirby-Bauer disc diffusion method and interpretation was done following Clinical and Laboratory Standard Institute (CLSI) guidelines, 2013. ESBL screening among E. coli and Klebsiella spp. isolates were done using ceftriaxone, cefotaxime, ceftazidime and cefpodoxime. The confirmation was done by phenotypic disc diffusion test (combined disc method) using ceftazidime (30µg) and ceftazidime plus clavulanic acid (30/10µg), and cefotaxime (30µg) and cefotaxime plus clavulanic acid (30/10µg) disc as per CLSI guidelines. RESULTS: A total of 451 samples showed significant bacteriuria with 365 (80.9%) E. coli, 17 (3.8%) Klebsiella pneumoniae and 3 (0.7%) Klebsiella oxytoca. Of 451 isolates, 236 (52.3%) were found MDR strains. By combined disk test, 33 (91.7%) E. coli and 3 (8.3%) Klebsiella spp. were found ESBL producers. CONCLUSION: Higher prevalence of ESBL producing E. coli and Klebsiella spp. was observed warranting prompt need of surveillance for effective management of such MDR strains.

Predictors of duration and treatment failure of severe pneumonia in hospitalized young Nepalese children.

BACKGROUND: Pneumonia in young children is still the most frequent cause of death in developing countries. We aimed to identify predictors for recovery and treatment failure in children hospitalized with severe pneumonia. METHODS: We enrolled 610 Nepalese children, aged 2 - 35 months from February 2006 to June 2008. Study participants were provided with standard treatment for pneumonia and followed up until discharge. Three multiple regression models representing clinical variables, clinical and radiological combined and all variables, including C-reactive protein (CRP) and viral etiology were used to assess the associations. RESULTS: The median age of study participants was 6 months with 493 (82%) infants and 367 (61%) males. The median time (IQR) till recovery was 49 (31, 87) hours and treatment failure was experienced by 209 (35%) of the children. Younger age, hypoxia on admission and radiographic pneumonia were independent predictors for both prolonged recovery and risk of treatment failure. While wasting and presence of any danger sign also predicted slower recovery, Parainfluenza type 1 isolated from the nasopharynx was associated with earlier resolution of illness. Gender, being breastfed, stunting, high fever, elevated CRP, presence of other viruses and supplementation with oral zinc did not show any significant association with these outcomes. CONCLUSION: Age, hypoxia and consolidation on chest radiograph were significant predictors for time till recovery and treatment failure in children with severe pneumonia. While chest radiograph is not always needed, detection and treatment of hypoxia is a crucial step to guide the management of hospitalized children with pneumonia.

Cyclospora Infection among School Children in Kathmandu, Nepal: Prevalence and Associated Risk Factors.

BACKGROUND: The intestinal coccidian protozoa Cyclospora cayetanensis has emerged as an important cause of parasitic diarrhea among children living in developing countries. This study aimed to determine the prevalence of Cyclospora among the school children of Kathmandu with reference to various associated risk factors. METHODOLOGY: A total of five hundred and seven stool samples from students between the age of 3-14 years, studying in 13 different schools in Kathmandu were collected during the study period (May-November, 2014) and processed at the Public Health Research Laboratory, Institute of Medicine, Kathmandu, Nepal. A modified acid fast staining technique (Kinyoun's method) was used to detect oocyst of Cyclospora from the formal-ether concentrated stool samples. RESULTS: Cyclospora was detected in 3.94% (20/507) of the stool samples examined. The prevalence was found to be highest among the students in the 3-5 year age group i.e. 10.15% (13/128), peaking during the rainy season (June-August). The detection rate was found to be significantly higher (p < 0.05) among children presenting with diarrheal symptoms, household keeping livestock and consumers of raw vegetables/fruits, showing a prevalence of 10.57% (11/104), 10.11% (9/89) and 7.25% (14/193) respectively. CONCLUSION: Consumption of untreated drinking water, fresh produce (raw fruits/vegetables) without proper washing and the presence of livestock at home were found to be predisposing factors for higher susceptibility of infection due to Cyclospora. This finding confirms the existence of a public-health issue with potentially serious consequences whereby children can be infected through exposure to oocysts in contaminated food and water and get ill as a result.