Burden of human metapneumovirus infections among children with acute respiratory tract infections attending a Tertiary Care Hospital, Kathmandu.

BACKGROUND: Acute respiratory infections (ARIs) are one of the most common causes of mortality and morbidity worldwide. Every year millions of children suffer from viral respiratory tract infections (RTIs) ranging from mild to severe illnesses. Human Metapneumovirus (HMPV) is among the most frequent viruses responsible for RTIs. However, HMPV infections and their severity among children have not been explored yet in Nepal. PURPOSE: Therefore, the study was focused on HMPV infections and other potential viral etiologies or co-infections using multiplex PCR among children attending Kanti Children's Hospital and assessed the clinical characteristics of the infections as well as found the co-infections. A hospital-based cross-sectional study was designed and a convenience sampling method was used to enroll children of less than 15 years with flu-like symptoms from both outpatients and inpatients departments over three months of the study period. RESULTS: HMPV infection (13.3%) was the most predominant infection among the different viral infections in children with ARIs in Kanti Children's Hospital. The HMPV was more prevalent in the age group less than three years (21.8%). Cough and fever were the most common clinical features present in all children infected with HMPV followed by rhinorrhea, sore throat, and wheezing. HMPV-positive children were diagnosed with pneumonia (42.9%), bronchiolitis (28.5%), upper respiratory tract infections (14.3%), and asthma (14.3%). The prevalence of HMPV was high in late winter (14.3%) followed by early spring (13.5%). CONCLUSIONS: This study provides the baseline information on HMPV and associated co-infection with other respiratory viruses for the differential diagnosis based on molecular methods and also the comparison of clinical presentations among the different respiratory syndromes.

Detection of NDM Variants (bla NDM-1, bla NDM-2, bla NDM-3) from Carbapenem-Resistant Escherichia coli and Klebsiella pneumoniae: First Report from Nepal.

Background: Increasing burden of carbapenem resistance among Enterobacterales is attributable to their ability to produce carbapenemase enzymes like metallo-beta-lactamase (MBL), Klebsiella pneumoniae carbapenemase (KPC), and OXA-type. This study aimed to determine the prevalence of carbapenemases and MBL genes ((bla NDM-1, bla NDM-1 and bla NDM-3) among E. coli and K. pneumoniae isolates. Methods: A total of 2474 urine samples collected during the study period (July-December 2017) were processed at the microbiology laboratory of Kathmandu Model Hospital, Kathmandu. Isolates of E. coli and K. pneumoniae were processed for antimicrobial susceptibility testing (AST) by disc diffusion method. Carbapenem-resistant isolates were subjected to Modified Hodge Test (MHT) for phenotypic confirmation, and inhibitor-based combined disc tests for the differentiation of carbapenemase (MBL and KPC). MBL-producing isolates were screened for NDM genes by polymerase chain reaction (PCR). Results: Of the total urine samples processed, 19.5% (483/2474) showed the bacterial growth. E. coli (72.6%; 351/483) was the predominant isolate followed by K. pneumoniae (12.6%; 61/483). In AST, 4.4% (18/412) isolates of E. coli (15/351) and K. pneumonia (3/61) showed resistance towards carbapenems, while 1.7% (7/412) of the isolates was confirmed as carbapenem-resistant in MHT. In this study, all (3/3) the isolates of K. pneumoniae were KPC-producers, whereas 66.7% (10/15), 20% (3/15) and 13.3% (2/15) of the E. coli isolates were MBL, KPC and MBL/KPC (both)-producers, respectively. In PCR assay, 80% (8/10), 90% (9/10) and 100% (10/10) of the isolates were positive for bla NDM-1, bla NDM-2 and bla NDM-3, respectively. Conclusion: Presence of NDM genes among carbapenemase-producing isolates is indicative of potential spread of drug-resistant variants. This study recommends the implementation of molecular diagnostic facilities in clinical settings for proper infection control, which can optimize the treatment therapies, and curb the emergence and spread of drug-resistant pathogens.

Plasmid-Mediated AmpC ß-Lactamase CITM and DHAM Genes Among Gram-Negative Clinical Isolates.

BACKGROUND: Antibiotic resistance mediated by the production of extended-spectrum ß-lactamases (ESBLs) and AmpC ß-lactamases is posing a serious threat in the management of the infections caused by Gram-negative pathogens. The aim of this study was to determine the prevalence of two AmpC ß-lactamases genes, bla CITM and bla DHAM, in Gram-negative bacterial isolates. MATERIALS AND METHODS: A total of 1151 clinical samples were obtained and processed at the microbiology laboratory of Annapurna Neurological Institute and Allied Science, Kathmandu between June 2017 and January 2018. Gram-negative isolates thus obtained were tested for antimicrobial susceptibility testing (AST) using Kirby-Bauer disk diffusion method. AmpC ß-lactamase production was detected by disk approximation method using phenylboronic acid (PBA). Confirmed AmpC ß-lactamase producers were further screened for bla CITM and bla DHAM genes by conventional polymerase chain reaction (PCR). RESULTS: Out of 1151 clinical specimens, 22% (253/1152) had bacterial growth. Of the total isolates, 89.3% (226/253) were Gram-negatives, with E. coli as the most predominant species (n=72) followed by Pseudomonas aeruginosa (n=41). In the AST, 46.9% (106/226) of the Gram-negative isolates were multidrug resistant (MDR). In disk diffusion test, 113 (50%) isolates showed resistance against cefoxitin, among which 91 isolates (83 by disk test and Boronic acid test, 8 by Boronic test only) were confirmed as AmpC ß-lactamase-producers. In PCR assay, 90.1% (82/91) and 87.9% (80/91) of the isolates tested positive for production of bla CITM and bla DHAM genes, respectively. CONCLUSIONS: High prevalence of AmpC ß-lactamase-producers in our study is an alarming sign. This study recommends the use of modern diagnostic facilities in the clinical settings for early detection and management which can optimize the treatment therapies, curb the growth and spread of the drug-resistant pathogens.

Extended spectrum beta-lactamase and metallo beta-lactamase production among Escherichia coli and Klebsiella pneumoniae isolated from different clinical samples in a tertiary care hospital in Kathmandu, Nepal.

BACKGROUND: Extended spectrum beta-lactamase (ESBL) and metallo beta-lactamase (MBL) production in Klebsiella pneumoniae and Escherichia coli are the commonest modes of drug resistance among these commonly isolated bacteria from clinical specimens. So the main purpose of our study was to determine the burden of ESBL and MBL production in E. coli and K. pneumoniae isolated from clinical samples. Further, the antimicrobial susceptibility patterns of E. coli and K. pneumoniae were also determined. METHODS: A cross-sectional study was conducted at Om Hospital and Research Centre, Kathmandu, Nepal by using the E. coli and K. pneumoniae isolated from different clinical samples (urine, pus, body fluids, sputum, blood) from May 2015 to December 2015. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Extended spectrum beta-lactamase production was detected by combined disc method using ceftazidime and ceftazidime/clavulanic acid discs and cefotaxime and cefotaxime/clavulanic acid discs. Similarly, metallo beta-lactamase production was detected by combined disc assay using imipenem and imipenem/ethylenediaminetetracetate discs. Bacteria showing resistance to at least three different classes of antibiotics were considered multidrug resistant (MDR). RESULTS: Of total 1568 different clinical samples processed, 268 (17.1%) samples were culture positive. Among which, E. coli and K. pneumoniae were isolated from 138 (51.5%) and 39 (14.6%) samples respectively. Of the total isolates 61 (34.5%) were ESBL producers and 7 (4%) isolates were found to be MBL producers. High rates of ESBL production (35.9%) was noted among the clinical isolates from outpatients, however no MBL producing strains were isolated from outpatients. Among 138 E. coli and 39 K. pneumoniae, 73 (52.9%) E. coli and 23 (59%) K. pneumoniae were multidrug resistant. The lowest rates of resistance was seen toward imipenem followed by piperacillin/tazobactam, amikacin and cefoperazone/sulbactam. CONCLUSIONS: High rate of ESBL production was found in the E. coli and K. pneumoniae isolated from outpatients suggesting the dissemination of ESBL producing isolates in community. This is very serious issue and can't be neglected. Regular monitoring of rates of ESBL and MBL production along with multidrug resistance among clinical isolates is very necessary.

Anti Japanese encephalitis virus IgM positivity among patients with acute encephalitic syndrome admitted to different hospitals from all over Nepal.

The Japanese encephalitis virus (JEV) infection is one of the major public health problems in Nepal because of its increasing disease morbidity and mortality. The main purpose of this study was to determine the anti-JEV IgM positivity among acute encephalitis syndromic cases from all over Nepal. The present study was conducted at National Public Health Laboratory, Kathmandu, Nepal from April 2015 to October 2015. A total of 671 (418 CSF and 253 serum) samples were collected from 625 patients with acute encephalitic syndrome, admitted to different hospitals from all over Nepal. IgM antibody capture enzyme linked immunosorbent assay (ELISA) was used for the detection of anti-JEV IgM positive cases. The rate of anti-JEV IgM positivity was found to be 21.12%. The majority of positive cases (50%) were from the age group below 15 years, with the highest numbers of cases occurring in September (55.30%). Among all the anti-JEV IgM positive cases, higher numbers of cases were males. Geographically, the highest numbers of anti-JEV IgM positive cases were recorded from Terai region. Similarly, largest numbers of anti-JEV IgM positive cases were reported from Kailai district followed by those from Kanchanpur. However, anti-JEV IgM positive cases were also reported from hill districts. Continuation of active surveillance and vector control measures, proper management of diagnostic facilities and expanded program of immunization in JE endemic areas should be strongly emphasized to reduce the endemicity of the disease.

Antibiotic resistance and biofilm production among the strains of Staphylococcus aureus isolated from pus/wound swab samples in a tertiary care hospital in Nepal.

BACKGROUND: The increasing drug resistance along with inducible clindamycin resistance, methicillin resistance and biofilm production among the strains of Staphylococcus aureus are present as the serious problems to the successful treatment of the infections caused by S. aureus. So, the main objectives of this study were to determine the antimicrobial susceptibility patterns along with the rates of inducible clindamycin resistance, methicillin resistance and biofilm production among the strains of S. aureus isolated from pus/wound swab samples. METHODS: A total of 830 non-repeated pus/wound swab samples were processed using standard microbiological techniques. The colonies grown were identified on the basis of colony morphology, Gram's stain and biochemical tests. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Detection of inducible clindamycin resistance was performed by D test, while detection of methicillin resistant S. aureus (MRSA) was performed by determination of minimum inhibitory concentration of oxacillin by agar dilution method. Similarly, detection of biofilm formation was performed by microtiter plate method. Strains showing resistance to three or more than three different classes of antibiotics were considered multidrug resistant. RESULTS: Total 76 samples showed the growth of S. aureus, among which 36 (47.4%) contained MRSA and 17 (22.4%) samples were found to have S. aureus showing inducible clindamycin resistance. Among the S. aureus isolated from outpatients, 41.9% were MRSA. Highest rates of susceptibility of S. aureus were seen toward linezolid (100%) and vancomycin (100%). Similarly, S. aureus isolated from 35 (46.1%) samples were found to be biofilm producers. Higher rate of inducible clindamycin resistance was seen among MRSA in comparison to methicillin susceptible S. aureus (MSSA). Similarly, higher rates of multidrug resistance and methicillin resistance were found among biofilm producing strains in comparison to biofilm non producing strains. CONCLUSIONS: The rate of isolation of MRSA from community acquired infections was found to be high in Nepal. Increased rate of inducible clindamycin resistance as compared to previous studies in Nepal was noted. So for the proper management of the infections caused by S. aureus, D test for the detection of inducible clindamycin resistance should be included in the routine laboratory diagnosis. Further, detection of biofilm production should also be included in the routine tests. Linezolid and vancomycin can be used for the preliminary treatment of the serious infections caused by S. aureus.