Biology subjects

Tuladhar, R.

Publications and source records attributed to Tuladhar, R..

5 recordsLinked to original sources

bioRxiv2026

Lytic bacteriophages against Salmonella Typhi as a potential alternative to antibiotics

IntroductionEnteric fever, being endemic with seasonal peaks in low- and middle-income countries, is a major health concern. Moreover, the rise in antibiotic resistance has exacerbated the situation. This study was undertaken to investigate the lytic bacteriophages against Salmonella Typhi with a potential for phage therapy. Materials and MethodsA hospital-based cross-sectional study was conducted from October 2023 to March 2024. Blood cultures were processed by the BACTEC automated culture system following standard microbiological techniques to isolate typhoidal Salmonella. Antibiotic susceptibility was tested by the modified Kirby-Bauer disc diffusion method. Lytic bacteriophages isolated by the double-layer agar method were assessed for their host range and lytic ability with spot and turbidimetric assays. ResultsOf the total 1054 blood specimens, 35 (3.2%) were positive for S. Typhi. All the isolates were susceptible to first-line antibiotics--ampicillin, chloramphenicol, and cotrimoxazole. The isolates were also sensitive to nalidixic acid (80%) as well as fluoroquinolones; ciprofloxacin (62.86%), levofloxacin (77.14%), and ofloxacin (80%). Fifteen lytic phages were isolated against S. Typhi Ty2 and CT18 strains. Four phages--vB_SaTy_ST2, vB_SaTy_ST7, vB_SaTy_ST17, and vB_SaTy_ST18--lysed all 35 clinical S. Typhi isolates. While vB_SaTy_ST17 and vB_SaTy_ST18 also lysed 7 out of 20 S. Paratyphi A isolates. Three phages (vB_SaTy_ST2, vB_SaTy_ST7, vB_SaTy_ST17) were tested against S. Typhi isolate S30. Individually, vB_SaTy_ST17 suppressed the growth for 13 hours, vB_SaTy_ST2 and vB_SaTy_ST7 for 10 hours. The phage cocktail vB_SaTy_ST2 + vB_SaTy_ST17 was the most effective, which extended the inhibition time to 15 hours. ConclusionThis study highlights the ongoing burden of enteric fever in Nepal and the increase in susceptibility of S. Typhi to nalidixic acid and fluoroquinolones. It also demonstrates the promising lytic potential of bacteriophages, particularly vB_SaTy_ST17 and the phage cocktail vB_SaTy_ST2 + vB_SaTy_ST17, against clinical S. Typhi, highlighting their potential as alternatives to antibiotics.

microbiology↗

bioRxiv2024

Prevalence of Class 1 integron in disease-causing carbapenem-resistant Escherichia coli in a tertiary hospital in Kathmandu, Nepal

BackgroundHigh prevalence of carbapenem resistance genes have been abundantly reported within clinical E. coli in Nepal but their co-prevalence with class-1 integron is poorly documented. These Class I Integrons play pivotal role in evolution and spread of antimicrobial resistance (AMR). Therefore, it is necessary to highlight their epidemiological significance, so this study determined the prevalence of Class 1 integrons within Carbapenem resistance Escherichia coli (CREc) population. MethodsFifty-two clinical E. coli with confirmed resistance to Meropenem were collected between January 2020-April 2022 in Kathmandu Model Hospital, and undergone antimicrobial susceptibility testing for 11 antimicrobial classes. Carbapenemase production was confirmed by eCIM and mCIM methods. Presence of Class I integron and carbapenemase genes, blaNDM-1, blaKPC, blaIMP, blaVIM, blaOXA-23, blaOXA-48, was confirmed by PCR method. ResultsThe result revealed 92.3% (n=48/52) of the isolates harbored intI1 gene. Among integron-positive population, 97.9% (n=47/48) were carbapenemase producers which were further identified as metallo {beta}-lactamase (MBL) producers (82.9%, n=39/47) and serine {beta}-lactamase (SBL) producer (17%, n=8/47). Carbapenemase gene blaNDM-1 (79.2%, n=38/48) was highly prevalent within integron-positive isolates, followed by blaOXA-23 (62.5%, n=30/48) and blaOXA-48 (54.2%, n=26/48). All integron-positive CREc isolates were resistant to minimum 16 antibiotics tested, but were susceptible to Colistin, Polymixin B, and Tigecycline. ConclusionIn conclusion, Class 1 integrons were highly prevalent within clinical CREc in Nepal, and their co-prevalence with other CRE gene is concerning due to possibility of wider spread. Therefore, an in-depth genomic analysis of Class 1 integrons is needed to understand the integron mediated AMR in high risk clones like CREc and inform intervention measures to stop spread of such pathogens.

microbiology↗

bioRxiv2023

Emergence of ISAba1-linked oxacillinase genes among carbapenem resistant Acinetobacter baumannii isolates in a tertiary cardiac center, Nepal

Insertion sequence contributes to the emergence of carbapenem resistance by dissemination of carbapenemase genes and providing promoter for their overexpression. This study aims to ascertain the occurrence of ISAba1-linked OXA carbapenemase genes and its relevance to carbapenem resistance level in Acinetobacter baumannii. This hospital based descriptive study was conducted at Shahid Gangalal National Heart Center, Kathmandu, Nepal. An overall of 1,291 clinical specimens received for routine culture and antibiotic susceptibility testing throughout the study period were included in this study. Identification of Acinetobacter baumannii was validated through detection of intrinsic blaOXA-51-like gene by polymerase chain reaction (PCR). Antibiotic susceptibility was tested by Kirby-Bauer disc diffusion approach and minimum inhibitory concentration (MIC) of meropenem was assessed through agar dilution method. Uniplex PCR assays were performed to detect genes encoding oxacillinases and ISAba1. Upstream association of insertion element, ISAba1 to oxacillinase genes was assessed through PCR mapping strategy using ISAba1F and OXA-51R/OXA-23R primers. Out of the 340 bacteria isolated, only 40 (11.8%) were Acinetobacter baumannii. All isolates were resistant against meropenem with MIC value ranging from 16-256 g/ml. blaOXA-23-like genes was present in every isolate but blaOXA-58 in just two isolates (5%). All isolates had ISAba1 either above blaOXA-23-like or blaOXA-51-like gene. Higher MIC90 value of meropenem (243.20 g/ml) was found in A. baumannii cluster with ISAba1-linked upstream to both blaOXA-23-like and blaOXA-51-like genes, thus depicting their eminent role to enhanced carbapenem resistance. Acinetobacter baumannii isolates with ISAba1-linked oxacillinase genes are rapidly emerging in clinical settings of Nepal. Thus, medical communities need to be prepared and enable targeted approaches for managing burgeoning problem of carbapenem resistance.

microbiology↗

bioRxiv2019

CRISPR/Cas9-based mutagenesis frequently provokes on-target mRNA misregulation

The introduction of insertion-deletions (INDELs) by activation of the error-prone non-homologous end-joining (NHEJ) pathway underlies the mechanistic basis of CRISPR/Cas9-directed genome editing. The ability of CRISPR/Cas9 to achieve gene elimination (knockouts) is largely attributed to the emergence of a pre-mature termination codon (PTC) from a frameshift-inducing INDEL that elicits non-sense mediated decay (NMD) of the mutant mRNA. Yet, the impact on gene expression as a consequence of CRISPR/Cas9-introduced INDELs into RNA regulatory sequences has been largely left uninvestigated. By tracking DNA-mRNA-protein relationships in a collection of CRISPR/Cas9-edited cell lines that harbor frameshift-inducing INDELs in various targeted genes, we detected the production of foreign mRNAs or proteins in [~]50% of the cell lines. We demonstrate that these aberrant protein products are derived from the introduction of INDELs that promote internal ribosomal entry, convert pseudo-mRNAs into protein encoding molecules, or induce exon skipping by disruption of exon splicing enhancers (ESEs). Our results using CRISPR/Cas9-introduced INDELs reveal facets of an epigenetic genome buffering apparatus that likely evolved to mitigate the impact of such mutations introduced by pathogens and aberrant DNA damage repair, and that more recently pose challenges to manipulating gene expression outcomes using INDEL-based mutagenesis.

molecular biology↗

bioRxiv2019

Biophotons: low signal/noise ratio reveals crucial events

We study the emission of photons from germinating seeds using an experimental technique designed to detect photons of extremely small intensity when the signal/noise ratio is low. We analyze the dark count signal in the absence of germinating seeds as well as the photon emission during the germination process. The technique of analysis adopted here was originally designed to measure the temporal complexity of astrophysical, sociological and physiological processes. The foundation of this method, called Diffusion Entropy Analysis (DEA), rests on Kolmogorov complexity. The updated version of DEA used in this paper is designed to determine if the signal complexity is generated by either non-ergodic crucial events with a non-stationary correlation function or by the infinite memory of a stationary but non-integrable correlation function or by a mixture of both processes. We find that dark count yields the ordinary scaling, thereby showing that no complexity of either kinds may occur in the absence of any seeds in the chamber. In the presence of seeds in the chamber anomalous scaling emerges, reminiscent of that found in neuro-physiological processes. However, this is a mixture of both processes and with the progress of germination the non-ergodic component tends to vanish and complexity is dominated by the stationary infinite memory. We argue that this may be a sign of quantum coherence that according to some authors is the important ingredient of cognition.

biophysics↗