Objective: To gauge the recent breadth of MDR E. coli along with antibiogram of carbapenemase producing (CP) E. coli among children from an institute which receives patients from all over Punjab. Methods: The bacterial strains of E. coli isolated from various specimens of patients were collected from April 2017 to August 2018 and processed using standard biochemical tests and API 20E system (bioMerieux). Phenotypic screening for CP E. coli was done by the modified Hodge test, whereas antibiotic susceptibility testing was done with Kirby-Bauer disc diffusion technique. Results: Total of 6,468 bacterial strains were isolated, out of which 1,552 (24%) were E. coli. Carbapenem resistance was observed in 245 (16%) strains, amongst which 113 (46%) were confirmed to be CP. E. coli isolated from males were higher as compared to females (p<0.05). Majority of the organisms were isolated from blood (37.2%) samples. The hospital discharged about 65% of patients, while 23% left against medical advice. Overall MDR amongst E. coli was 93.26%. Colistin sulphate (15.9%) and nitrofurantoin (16.8%) showed the most efficacy followed by amikacin (15%) and fosfomycin (10.6%). Conclusion: The isolation of high number of MDR E. coli amongst the paediatric patients is worrisome, which could serve as a potential source of horizontal genes transfer to other genera. doi: https://doi.org/10.12669/pjms.36.2.928 How to cite this:Nosheen S, Bukhari NI, Ejaz H, Abbas N. Antibiogram and recent incidence of multi-drug resistant carbapenemase producing Escherichia coli isolated from paediatric patients. Pak J Med Sci. 2020;36(2):---------. doi: https://doi.org/10.12669/pjms.36.2.928 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

The evolution of NDM genes ( bla NDM ) in E. coli is accounted for expansive multidrug resistance (MDR), causing severe infections and morbidities in the pediatric population. This study aimed to analyze the phylogeny and mutations in NDM variants of E. coli recovered from the pediatric population. Carbapenem-resistant clinical strains of E. coli were identified using microbiological phenotypic techniques. PCR technique used to amplify the bla NDM genes, identified on agarose gel, and analyzed by DNA sequencing. The amino acid substitutions were examined for mutations after aligning with wild types. Mutational and phylogenetic analysis was performed using Lasergene, NCBI blastn, Clustal Omega, and MEGA software, whereas PHYRE2 software was used for the protein structure predictions. PCR amplification of the bla NDM genes detected 113 clinical strains of E. coli with the contribution of bla NDM-1 (46%), bla NDM-4 (3.5%), and bla NDM-5 (50%) variants. DNA sequencing of bla NDM variants showed homology to the previously described bla NDM-1 , bla NDM-4, and bla NDM-5 genes available at GenBank and NCBI database. In addition, the mutational analysis revealed in frame substitutions of Pro60Ala and Pro59Ala in bla NDM-4 and bla NDM-5, respectively. The bla NDM-1 was ortholog with related sequences of E. coli available at GenBank. The phylogenetic analysis indicated that the NDM gene variants resemble other microbes reported globally with some new mutational sites.

Background: The emergence of AmpC beta-lactamase producing Enterobacter cloacae becomes a serious nosocomial menace due to wider resistance. The study aimed to know the existence of these superbugs in the hospital settings and to report the current trends in their antibiotic resistance. Methods: We chose a tertiary care pediatric hospital for this cross-sectional study and processed 27,000 clinical specimens for the isolation of E. cloacae using routine microbiological procedures. A total number of 96 E. cloacae isolates from various sources were screened for AmpC production with cefoxitin (30 μg) and confirmed by inhibitor based technique. The antibacterial drug resistance studied against various groups of antibiotics in vitro. Results : Boronic acid inhibitor based method revealed 63 (65.6%) pathogens as AmpC beta-lactamase producing E. cloacae. Most of the infected patients with AmpC producing E. cloacae were neonates (34; 54.0%) and infants (11; 17.5%). The primary source of AmpC producing E. cloacae was blood (43; 68.3%), and they were frequently distributed in the neonatal nursery unit (33; 52.4%) and medical ward (13; 20.6%). All of these bugs showed a high level of resistance (100%) against the co-amoxiclav and cephalosporin group.The organisms exhibited less resistance to levofloxacin, imipenem and colistin sulphate as 23 (36.5%), 20 (31.7%) and 17 (27.0%), respectively. Conclusion: The consistent emerging threat of Amp C harbouring E. cloacae could disseminate AmpC genes in other genera of the bacteria which lead to the therapeutic failure and leave the doctors with limited treatment options of levofloxacin, imipenem and colistin sulphate. Bangladesh Journal of Medical Science Vol.19(4) 2020 p.632-637