Escherichia coli Genome sequencing. Escherichia coli

The ever increasing incidences of antimicrobial resistance in both domestic animals and human beings are well documented worldwide. The situations are becoming more complicated, as the development of resistance spreads to wild animals. Reports of Multi Drug Resistant isolates driving from the wild animals at the whole genome level are limited. An Asiatic female elephant aged 55 years from which multi drug resistant Escherichia coli was isolated from the urine sample, was under private ownership. The isolate was resistant against 27 antimicrobial agents. The captive Asiatic elephant was passing blood in urine, dull, dehydrated and anorectic. The elephant was kept outdoors and moving from one place to another on foot at the wish of the owner. The elephant was feed by public in the market roads in addition to the diets given by the caretaker. The elephant found be dull, anorectic, dehydrated and passing scanty volume of urine. At least 20 ml of midstream voided urine was collected aseptically in sterile containers in June 2023.Urine samples were inoculated in buffered water and were incubated. The culture was inoculated in McConkey agar and incubated . Pink color colonies were further streaked on eosin methylene blue agar and were incubated. Confirmation of purified colonies on eosin methylene blue agar and antimicrobial susceptibility pattern was done by using automated bacterial identification by both oxidation-reduction indicator and turbidity growth detection and antimicrobial susceptibility test system from Becton and Dickson. The Escherichia coli isolate was stored at deep freezer in LB broth with supplementation of glycerol. Antimicrobial susceptibility testing was carried against 27 antibiotic.Genomic DNA from E coli cultures were extracted and quantified with DNA HS assay kit. The library was prepared using the Illumina trueseq Nano DNA Library kit protocol according to the manufactures instructions. The gDNA was fragment and tagged with adapter sequences using a transposase enzyme in a process known as tagmentation. A size selection clean-up was done using AMPure beads and normalized to ensure equal representation of all libraries. The normalized libraries were pooled, denatured and loaded on the Illumina Novaseq 6000 platform with an out of 50bp chemistry.