Metallomics dataset of clinical isolates of Klebsiella pneumoniae exposed to varying concentration of Human Plasma

Description

The dataset consists of growth data (seeded at high density, i.e., 10^7 cell/ml) and ICP-MS intracellular metal ion profile of 7 antibiotic resistant clinical bloodstream isolates of K. pneumoniae and ATCC strain (33495 ™) grown in varied h-plasma concentration (0, 5, 10, 25, 55 and 100%) supplemented in MHB for 4 hrs.

Steps to reproduce

Bacterial culture, growth evaluation, and pellet generation Pure colonies of K. pneumoniae were obtained by incubating bloodstream-isolated 7 clinical strains of K. pneumoniae and ATCC strain (33495 ™) on Mueller Hinton agar plates at 37 °C overnight, followed by an additional overnight preparatory broth culture at 37 °C in MHB with shaking at 220 rpm at 37°C, confirmed using MALDI-TOF MS. The secondary cultures were then inoculated (1: 500 v/v) in fresh MHB and incubated under aerobic conditions at 37°C until the mid-exponential phase of growth (~0.4-0.6 OD600nm post-inoculation). 1*107 cells of exponentially grown K. pneumoniae cells (different clinical isolate) were collected using centrifugation at 6000 RPM and seeded in 1 ml of MHB, h-plasma, and different concentrations (1, 5, 10, 25 and 55%) of h-plasma in MHB, incubated for 4 hrs., followed by plating (10ul: diluted in different ratios) on MH agar plate for CFU/ml calculation. Furthermore, incubated cells were collected as a pellet using centrifugation at 10,000 RPM, followed by 2 washes using PBS containing 5 mM ethylenediaminetetraacetic acid, and twice with PBS to remove residual MHB/h-plasma content from the pellet [11] and subjected to sample preparation for metallomics. Sample preparation for ICP-MS Bacterial pellets were desiccated for 6 hrs. at 95°C and then digested in 250 μL 65% (vol/vol) HNO3 at 95°C for 20 minutes. Samples were centrifuged at 18,000g for 25 min, and supernatant(200 μL) was diluted to a final volume of 1.5 mL using MilliQ-H2O (ICP-MS grade). Elemental content was quantitatively analysed using ICP-MS XSERIES 2 Thermofisher Scientific using the parameters mentioned in “Acquisition Method Supplementary file 2” and Element Standard (Element Standard Curves Supplementary file 3). Data Analysis Based on colony count (CFU/ml) data, the impact of h-plasma (in different concentrations, i.e., 0, 1, 5, 10, 25, 55 and 100% in MHB) on the growth of K. pneumoniae strain was assessed, and bacterial strains were divided into 2 groups, i.e., h-plasma sensitive and insensitive, consisting of 4 strains each group. Next, the individual metal ion concentration/cell (total concentration of metal divided by CFU/ml for each strain) for each group of bacterial strains was plotted w.r.t h-plasma concentration and analysed to shortlist/identify metal ion profiles reflecting/associated with bacterial growth. The data is presented as Mean ± SD of the concentration of metal ions from each strain in a group. For statistical analysis, GraphPad Prism 10.1.0 software was used to analyse the data. Multiple unpaired T-test was used to analyse the results. For all tests, a P-value of <0.05 was considered to indicate statistical significance.

Institutions

• Translational Health Science and Technology Institute

Categories

Funders

• Department of Biotechnology, Ministry of Science and Technology Govt. of India

  Grant ID: BT/PR52308/BMS2/156/3/2024

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