Objectives: Blood stream infections (BSIs) are potentially fatal healthcare associated infections (HAIs). The COVID pandemic had a huge impact on hospital processes and health care outcomes. We studied the impact of COVID pandemic on the prevalence of ICU acquired BSIs in our established Indian HAI surveillance network. Material and methods: This study included adult patients from ICUs in AIIMS HAI network that conducted BSI surveillance in COVID and non-COVID ICUs during and before the pandemic periods. Of the 40 hospitals in the network, ICUs from 16 hospitals conducted HAI surveillance in COVID ICUs and were included for the purposes of this study. Hospitals identified BSI and reported clinical and microbiological data to the network as per established and previously published protocols. Results: A total of 1,863 events of BSI were identified during pre-pandemic and 2,571 events during pandemic period. During the pandemic, 557 (21.6%) were reported from COVID ICUs. The BSI rate during the pandemic was 8.9/1,000 patient days, and during pre-pandemic 5.4/1,000 patient days (P < 0.01). The central line associated BSI (CLABSI) rates increased from 9.2/1,000 central line days during the pre-pandemic period to 11.3/1,000 central line days during pandemic. Conclusion: An increase in BSIs during the COVID pandemic was observed, which may be attributed to increased susceptibility of the patients or suboptimal infection control practices in COVID ICUs.

Spontaneous bacterial peritonitis (SBP) is a life-threatening complication of idiopathic nephrotic syndrome (INS) in children. Evidence on the optimal duration of antibiotics in this population is limited. We investigated whether 5-day antibiotic therapy is non-inferior to 7-day therapy for SBP in children with INS. In this single-center, open-label, randomized controlled non-inferiority trial, 50 children aged 1-14 years with INS and SBP were allocated 1:1 to intravenous ceftriaxone for 5 days (Group A, n = 25) or 7 days (Group B, n = 25). The primary outcome was composite clinical response (resolution of fever, abdominal pain, and tenderness) at the end of treatment, analyzed by intention-to-treat. The pre-specified non-inferiority margin was -15%. Secondary outcomes included bacteriological cure, cytological response, and the need for additional antibiotics. Fifty children [median age 50 (30.8-92) months] were randomized. Composite clinical response was achieved in 25/25 (100%) in Group A versus 22/25 (88%) in Group B (risk difference 12%, 95% CI -3.5% to 30.0%; P = 0.23); the lower CI bound exceeded the -15% non-inferiority margin, establishing non-inferiority. Bacteriological cure was achieved in 96% of both groups. Cytological response (64% vs 72%; P = 0.54) was inconclusive for non-inferiority. Additional or modified antibiotic therapy was required in 16% versus 20% (P = 1.00). Median hospitalization was comparable (10 vs 11 days; P > 0.05). No ceftriaxone-related adverse events were recorded. Five-day intravenous ceftriaxone is non-inferior to 7-day therapy for SBP in children with INS, with respect to composite clinical response; the secondary outcome of cytological response was inconclusive for non-inferiority.

Background Carbapenem- and colistin-resistant Klebsiella pneumoniae poses a major challenge to critical care due to limited therapeutic options. Combined with hypervirulence, the infections caused by multi-drug-resistant isolates become exceptionally difficult to treat, often resulting in prolonged illness and higher mortality. Methods We analysed 65 clinical isolates of carbapenem and colistin resistant K. pneumoniae using whole genome sequencing to characterize resistance, virulence, and associated mobile genetic elements that may facilitate horizontal gene transfer. Results The isolates represented 17 sequence types, with ST147, ST231 and capsular loci KL64 and KL51 being the most prevalent. Pan-genome analysis revealed high genetic diversity with an open genome structure. Resistance genes were widely distributed where carbapenem resistance was primarily mediated by NDM-1 or OXA-232 in combination with mutations in OmpK35/36 porin. Colistin resistance was mostly associated with mutations in mgrB, pmrB, or crrB genes though 30% of phenotypically resistant isolates lacked known determinants, suggesting the presence of additional unknown mechanisms. Virulence factors included frequent detection of yersiniabactin with aerobactin (iucA) and hypermucoidy loci (rmpA) in subset of isolates. Conclusion The co-occurrence of resistance and virulence determinants in multiple genomes suggests that highly pathogenic and multidrug resistant K. pneumoniae strains are already circulating in India. The detection of multiple IncF and Col-type plasmids, known to facilitate the mobilization of antimicrobial and virulence genes, further highlights the potential for future convergence events. Collectively, this study provides a genomic snapshot and valuable baseline for India, emphasising the importance of continued surveillance to monitor and contain the emergence of high-risk convergent lineages.

Necrotizing soft tissue infections (NSTIs) carry 10 to 30% mortality. Current empirical antimicrobial guidance derives almost entirely from Western cohorts dominated by Streptococcus pyogenes and aerobic-anaerobic consortia, yet whether this microbial paradigm applies to tropical, high-antimicrobial-pressure settings has not been tested with culture-independent methods. We did a prospective cohort study of 169 patients with intraoperatively confirmed NSTI at a North Indian tertiary center (2021 to 2024). Wound tissue underwent aerobic and anaerobic culture, QIIME2-based 16S rRNA gene amplicon sequencing (V3-V4), and targeted SYBR Green quantitative PCR (qPCR) for Acinetobacter baumannii and S. pyogenes . The wound microbiota was overwhelmingly Gram-negative and polymicrobial, anchored by A. baumannii (culture, 33.7%; metagenomics, 49.1%; qPCR, 37.9%), Escherichia coli (32.0%), and Klebsiella pneumoniae (20.7%); S. pyogenes contributed only 4.7% of culture-positive cases. Polymicrobial wounds had higher Shannon diversity (2.59 versus 2.33; P = 0.048) and discrete community composition (PERMANOVA R 2 = 0.511; P = 0.010). Culture-metagenomics agreement ranged from almost perfect for Escherichia (κ = 0.849) to slight for Streptococcus (κ = 0.131). North Indian NSTIs present a microbial picture distinct from the Western paradigm, with implications for empirical therapy.

BACKGROUND:Necrotising soft tissue infections (NSTIs) are fulminant infections requiring rapid diagnosis and intervention. Conventional methods, such as anaerobic culture, are slow and insensitive for clostridial pathogens. OBJECTIVES:To develop and evaluate a multiplex PCR assay targeting three clinically relevant clostridia directly in NSTI samples. METHODS:A multiplex conventional PCR targeting the cpa (Clostridium perfringens), csa (Clostridium septicum), and 16S rRNA (Paeniclostridium sordellii) genes was optimised and applied to tissue/pus samples from 87 clinically diagnosed NSTI patients. Anaerobic culture served as the reference. Sensitivity, specificity, and diagnostic turnaround times were compared. RESULTS:Seven patients (8.0%, 95% CI 3.3-16.2) were positive for clostridia by PCR, compared with four (4.6%, 95% CI 1.3-11.4) by culture. PCR identified three additional culture-negative cases. While McNemar's test did not show statistical significance (p = 0.25), PCR achieved a sensitivity of 100% (95% CI 39.8-100.0), specificity of 96.4% (95% CI 89.8-99.2), and a negative predictive value of 100%. Turnaround time was 3-4 h, compared with 48-96 h for culture. Both patients with monomicrobial C. septicum NSTI succumbed rapidly, highlighting its prognostic importance. CONCLUSION:This is the first report of a multiplex PCR assay for direct detection of C. perfringens, C. septicum, and P. sordellii in NSTIs. The assay enhances detection, reduces turnaround time, and provides clinically relevant species-level identification. Multiplex PCR may serve as a valuable adjunct to culture in early NSTI diagnostics and could be expanded to include additional anaerobic pathogens.

Introduction Amplicon sequencing is a targeted approach used to assess the diversity of microbial communities by amplifying and sequencing a specific genetic locus from DNA. QIIME2 is one of the most prevalent methods for metagenomics analysis due to its plugin-based design wherein distinct modules can be utilized to perform specific functions. However, QIIME2 data input, and plugin utilization is cumbersome to navigate. Previous amplicon pipelines also lack host depletion and statistical biomarker identification modules from upstream and downstream analysis. Methods To this effect, we assembled a simple and customizable Zenity based GUI workflow for analysing amplicon data with Automating Microbial Community Analysis (AMCA). The analysis integrates key attributes of amplicon analysis: host depletion with Bowtie2 and biomarker prediction by LEfSe. The bash-based analysis guides and allows the user to select filtering parameters based on intermediate results while minimizing the need to navigate command-based plugins. Results The outputs from the AMCA workflow include the filtered and host-depleted raw sequencing data, taxonomic abundances, alpha and beta diversity indices, alpha rarefaction analysis, phylogenetic tree (rooted and unrooted) and significant features which explain key microbial differences between conditions/classes of the experiment. The implementation of the designed workflow has been tested on a pilot study based on amplicon sequencing in 100 samples from patients of Chronic Kidney Disease and healthy controls. The exploratory LEfSE analysis revealed key taxa Streptococcus, Bacteroides and Faecalibacterium to vary between disease and control conditions. The source code related to the analysis can be assessed from the Github repository at https://github.com/Nitika-Rana/AMCA. Conclusion The study delivers an efficient, user-friendly, and customizable workflow for amplicon analysis, simplifying QIIME2 execution while enabling host depletion and biomarker characterization.

The escalating threat of antimicrobial resistance (AMR), particularly among gram-negative pathogens, necessitates the development of novel therapeutic agents. Tridecaptins, a class of non-ribosomally synthesized lipopeptides with a novel mode of action, have garnered renewed interest in the fight against AMR. Our group previously identified tridecaptin M, a compound with a promising safety profile, prompting further investigation into its efficacy and preclinical characteristics. Here, we show that tridecaptin M exhibits potent activity against multidrug-resistant (MDR) Klebsiella pneumoniae and Escherichia coli without cross-resistance to colistin. It effectively inhibits biofilm formation and disrupts 50% of established biofilm at 10 µg/mL. Tridecaptin M demonstrates a favorable safety profile, as it does not inhibit the cardiac hERG channel and shows minimal interaction with cytochrome P450 enzymes, with no IC₅₀ detected up to 44.6 µg/mL. In vivo toxicity studies via subcutaneous administration confirm its safety up to 600 mg/kg, whereas intravenous administration reveals acute toxicity at ≥30  mg/kg, with biochemical evidence of skeletal muscle, cardiac, and hepatic involvement. In mouse infection models using a colistin-resistant MDR strain of K. pneumoniae reveal the in vivo potential of tridecaptin M and a dose-dependent efficacy at 10 mg/kg, 20 mg/kg, 50 mg/kg, and 100 mg/kg doses, showing a non-linear relationship. Tridecaptin M is metabolized by liver microsomes, with low clearance, and pharmacokinetic analysis in rats indicates favorable attributes, with a terminal half-life (T₁/₂) of 3.65 h intravenously and 8.81 h subcutaneously. Collectively, these data support the continued preclinical development of tridecaptin M as a promising candidate for treating severe gram-negative infections.

BACKGROUND:Extended-spectrum beta-lactamases (ESBLs), particularly in Enterobacterales, are major contributors to the growing problem of antibiotic resistance in Gram-negative bacilli. This guidance document provides an overview of the epidemiology, identification, and clinical management of infections caused by Extended-Spectrum Beta-Lactamase-producing Enterobacterales (ESBL-E). This guideline adds on the previously issued Indian Council of Medical Research (ICMR) guidelines on carbapenem-resistant organisms (CROs) and intended to support clinicians in making evidence-based decisions regarding the diagnosis and treatment of ESBL infections, thereby promoting effective patient management and antimicrobial stewardship. OBJECTIVES:The purpose of this guidance document aims to assist clinicians in selecting not only the right antibiotics to diagnose ESBLs, but also right tests to diagnose ESBLs, for effective management of ESBL infections. CONTENT:This guidance document highlights the importance of early and accurate identification of ESBL -producing Enterobacterales, outlines clinical syndromes that may require empirical antibiotic coverage for ESBLs, and offers guidance on appropriate de-escalation strategies. Additionally, it emphasizes on the optimal use of use of newer beta-lactam/beta-lactamase inhibitor (BLBLI) combinations, such as cefepime-enmetazobactam and ceftolozane-tazobactam, and also emphasizes on avoiding important drugs like ceftazidime-avibactam for ESBL -producing Enterobacterales.

OBJECTIVE:Definitive guidance regarding the duration of antibiotics for neonatal sepsis is lacking. We hypothesised that a 7-day antibiotic course is non-inferior to a 14-day course for treating culture-proven sepsis. DESIGN:Randomised, controlled, non-inferiority trial with masked outcome assessment in eight centres in a low and middle-income country. PATIENTS:Neonates with a birth weight (BW) ≥1000 g and blood culture-proven sepsis were randomised on day 7 of sensitive antibiotic therapy provided sepsis had clinically remitted. EXCLUSIONS:Staphylococcus aureus or fungal sepsis, and infections requiring prolonged antibiotics. We planned to enrol 350 per group, assuming 10% rate of primary outcome, +7% non-inferiority margin, one-sided 5% alpha, 90% power, 10% loss to follow-up. INTERVENTION:7 days (no further treatment); comparison: 14 days (7 days postrandomisation). OUTCOMES:Primary: relapse (definite or probable) within day 21 postantibiotic completion. SECONDARY OUTCOMES:composite of mortality or definite/probable/secondary sepsis and duration of hospitalisation. One interim analysis (per protocol (PP)) was planned. RESULTS:126 and 135 subjects were recruited in 7-day and 14-day groups, respectively, with mean (SD) birth weight (BW) 2250.9 (741.1) and 2187.8 (718.8) g. The trial was terminated early, based on interim PP analysis. 2/125 and 6/130 subjects had the primary outcome in 7-day and 14-day groups, respectively (risk difference (RD)=-3.0% (99.5% CI -9.2%, +3.1%), below non-inferiority margin). The composite secondary outcome also favoured the 7-day regimen (RD: -3.7% (99.5% CI -12.4% to +5.1%)). Duration of hospitalisation was shorter in 7-day group (median difference: -4 days (95% CI -5 to -3)). CONCLUSIONS:A 7-day course of antibiotics may be non-inferior to a 14-day course for uncomplicated bacterial neonatal sepsis. TRIAL REGISTRATION NUMBER:NCT03280147.

Background: Mothers delivering preterm are very often exposed to antibiotics in the peripartum period. We hypothesize this may select bacteria bearing antibiotic resistance genes (ARGs) in the breast milk and be transmitted to the neonate's mouth while feeding. This study aimed to determine the prevalence of ARGs coding for extended-spectrum beta-lactamases (ESBLs) (including AmpC beta-lactamases [ACBLs]) and carbapenemases in breast milk and neonatal oral swab samples of preterm mother-infant pairs, the concordance of ARGs between paired samples, and risk factors of ARGs coding for ESBLs and carbapenemases. Methods: Breast milk and oral swab samples were obtained aseptically from 100 preterm mother-infant pairs (gestation 28-34 weeks) by postpartum day 10. Multiplex PCR was used to detect 15 common ARGs in these samples. Potential risk factors of the presence of any ARG coding for ESBLs or carbapenemases in breast milk and oral swab samples were studied. Results: The commonest ARGs for ESBLs, ACBLs, and carbapenemases in breast milk were blaSHV (28%), blaCIT (33%), and blaIMP (49%), respectively; and oral swabs blaCTX-M1 (30%), blaCIT (58%), and blaIMP (24%), respectively. ARGs common to breast milk and oral swabs included blaCIT (13%), blaIMP (10%), blaCTX-M-1 (9%), and blaSHV (6%). Formula milk intake was associated with less oral carbapenemase ARGs. Conclusion: ARGs for ESBLs and carbapenemases are highly prevalent in preterm breast milk and oral swabs.

BackgroundTo investigate the geospatial epidemiology, clinical features, treatment patterns, and antimicrobial resistance (AMR) trends of Stenotrophomonas maltophilia bloodstream infections (BSIs) in Indian intensive care units (ICUs) participating in a standardized healthcare-associated infection (HAI) surveillance program from 2017 to 2024.MethodsThis retrospective, multicentric study analyzed surveillance data from 54 ICUs across India. Standardized HAI definitions and protocols were applied to characterize infection types, clinical outcomes, and antimicrobial susceptibility.ResultsA total of 271 S. maltophilia isolates were identified, with the highest burden in 2023–24 (n = 76, 28.0%). Central line-associated BSIs (CLABSIs) predominated (64.9%), though their proportion decreased over time, with non-CLABSIs rising from 7.4% (2017–18) to 42.1% (2023–24). Mortality was highest in secondary BSIs (60%), followed by CLABSIs (50.3%) and non-CLABSIs (36.4%). The median ICU stay for CLABSI patients was 21 days. No significant associations were observed between infection type and time to infection or length of stay. High resistance was observed to tobramycin (92%), amikacin (80%), and piperacillin-tazobactam (70%), while trimethoprim-sulfamethoxazole (64.7–94.7%), levofloxacin (93%), and minocycline (94.1%) retained activity.ConclusionS. maltophilia represents a significant ICU pathogen in India, underscoring the urgent need for genomic surveillance and resistance-guided therapeutic strategies.

Aim We aimed to understand the host and microbe interactions at the time of infection and inflammatory response in amoebic liver abscess (ALA) patients based on toll-like receptor (TLR) expression (mRNA), cytokine and IgG subtypes levels. Methods and results Liver aspirates from 100 ALA patients and 11 liver autopsy samples were used as negative controls. Blood samples from 100 ALA and 41 healthy individuals were collected. mRNA expression of TLR 1 to 9 genes was measured using reverse transcriptase polymerase chain reaction (RT-PCR). Serum cytokines level was quantified by flow cytometry. In-house ELISA for the analysis of IgG and its subtypes in the serum samples was performed. A total of 7 TLR genes (TLR1, TLR2, TLR4, TLR6, TLR7, TLR8 and TLR9) and 6 TLR genes (TLR1, TLR2, TLR3, TLR4, TLR5 and TLR8) were found to be elevated in liver aspirates and PBMCs respectively. Increased serum cytokine levels were observed in ALA patients vs. healthy controls. Interestingly, a significant increase in IgG and its subtypes (IgG1, IgG3 and IgG4) was found in the serum of ALA patients. Conclusion Increased levels of TLR, pro- and anti-inflammatory cytokines, IgG and its subtypes, are possibly linked with early-stage infection in ALA patients. Impact statement The role of TLRs in association with ALA might provide insights into new therapeutic strategies.

Single-particle and collective excitations in 116Sb are studied using the reaction 115In(alpha, 3n) 116Sb at abeam energy of 40 MeV and investigated using gamma -ray spectroscopic techniques. The existing level scheme is extended with the observation of several new transitions. The previously known band structures are extended to higher spins. Measurements of the directional correlation from oriented states (DCO) ratio and polarization asymmetry of the observed gamma rays are carried out to assign the spin-parities of the excited levels, populated in the present work. New band structures are interpreted as being due to the coupling of the valence particles with the 2p-2h intruder states of the Sn core. Further, the collective band structures are discussed on the basis of their rotational properties, total Routhian surface calculations, and the triaxial projected shell model approach.

PURPOSE:This study aims to identify the distribution of different clostridial species in human infections, sample types, and their antimicrobial susceptibility profile using the E-test method. MATERIALS AND METHODS:The samples were processed as per the standard bacteriological techniques. Automated anaerobic gas evacuation-replacement system was used to create anaerobiosis (85 % N2, 10 % CO2, 5 % H2). Antibiotic susceptibility of isolated Clostridium species was performed using gradient diffusion (E strip) method against metronidazole, clindamycin, and penicillin. RESULTS:On antibiotic susceptibility testing by E-test, we have found 96.77 % susceptibility to metronidazole, 90.32 % susceptibility to penicillin, and 74.19 % susceptibility to clindamycin. CONCLUSION:It can be inferred that the E-test, when performed with the right inoculum, and when given the adequate period of incubation, could be used to reliably perform AST in Clostridium species. It produces a reliable report within 96 h of sample processing.

OBJECTIVES:To estimate the profile of non-central line-associated primary bloodstream infections (non-CLABSIs) in intensive care units of Indian hospitals participating in the standardized health-care-associated infection surveillance program. METHODS:This is a multicentric, network-based, prospective surveillance study conducted in 180 individual intensive care units of 47 Indian tertiary care hospitals that were part of the Health-Care-Associated Infection Surveillance Network between May 2017 and April 2024. The non-CLABSIs were defined, monitored, and observed using standardized definitions and surveillance protocols (www.haisindia.com). RESULTS:A total of 7092 laboratory-confirmed non-CLABSI cases and 30,74,954 patient days from 2017 to 2024 were recorded. The overall pooled non-CLABSI rate was 2.3 per 1000 patient days. Gram-negative isolates were the most predominant (5240/7659; 68.4%), including Klebsiella spp. (1766/5240; 33.7%), Acinetobacter baumannii (1613/5240; 30.8%), and Escherichia coli (582/5240; 11.1%). Gram-positive isolates (1728/7659; 22.6%) predominantly included Staphylococcus aureus (953/1728; 55.1%) and Enterococcus spp. (747/1728; 43.2%). Carbapenem resistance was common in Gram-negative infections, particularly in A. baumannii (1253/1554; 80.6%) and Klebsiella spp. (1209/1697; 71.2%). Among Gram-positive, S. aureus exhibited a high level of resistance to methicillin (529/748; 70.7%). CONCLUSIONS:This surveillance study underscores the need to expand infection prevention and control strategies to include non-device-associated risk factors. This will lead to the formulation of comprehensive infection prevention and control programs, mitigating the burden and clinical outcomes of non-CLABSI.

BACKGROUND AND OBJECTIVES:Alterations in gut microbiota have been linked to pathophysiology of immune-mediated diseases like multiple sclerosis (MS). This study was undertaken to characterise the gut microbiome profile in North Indian MS patients and to evaluate gut health using biomarkers like zonulin (intestinal permeability) and calprotectin (intestinal inflammation). METHODS:84 Patients with relapsing-remitting MS patients (RRMS) of 18-75 years of age with an expanded disability status scale (EDSS) score less than or equal to 5.5 and 106 healthy controls (HC) were recruited for the study. Gut microbiota was sequenced using Illumina MiSeq. Clinical, demographic, anthropometric, and dietary details were recorded. Sandwich ELISA was used to quantify serum zonulin and fecal calprotectin levels. RESULTS:MS patients had lower alpha microbial diversity, while distinct beta diversity metrics were observed in MS and HC. Firmicutes was found to be the most abundant phylum in both groups with significant enrichment in MS than HC. In MS, significant depletion of commensal bacterial species like Faecalibacterium prausnitzii, Monoglobus pectinilyticus, and Bacillus species indicated gut dysbiosis. These alterations influenced the prevalence and functioning of metabolic pathways. Therefore, pathways involved in biosynthesis of long-chain fatty acids (LCFA) were significantly enriched in MS than HC, while generation of short-chain fatty acids were predominant in HC. In addition, high zonulin, without an increase in calprotectin levels was observed in MS patients. CONCLUSIONS:RRMS patients in North India have a decreased microbial diversity in terms of depletion of commensals. The dominance of LCFA generating pathways in MS patients might have triggered the proinflammatory reactions, that are possibly linked to the development of a highly permeable/leaky gut in MS.

PURPOSE:Increase in the incidence of colistin-resistant K. pneumoniae (ColR) necessitates the study of risk factors responsible for its emergence. The current study aims to investigate the risk factors and clinical outcomes in ColR patients. MATERIAL AND METHODS:A retrospective study was conducted at a tertiary care hospital in India over 18 months (March 2021 to August 2022). We included patients >18 years (41 male, 38 female), with ColR and colistin-intermediate (ColI) strains. These patients were assessed for demographics, underlying diseases, antimicrobial treatment, clinical scores (APACHE II, SOFA), and outcome. RESULTS:The presence of a central venous catheter (CVC) was a significantly associated risk factor for ColR infections (p = 0.011). Additionally, prior antibiotic exposure within 3 months (p = 0.049) and CVC presence (p = 0.034) were associated with non-survival among ColR patients. Higher APACHE II and SOFA scores were associated with mortality among ColI patients. The presence of CCI >3 (p = 0.040) and previous history of ICU stay (p = 0.032) emerged as significant factors associated with the isolation of ColR K. pneumoniae among patients with previous history of colistin/polymyxin intake. CONCLUSION:The study highlights the risk factors associated with ColR K. pneumoniae. These findings underscore the need for prudent antimicrobial stewardship.