Effect of dilution on sedimentational separation of bacteria from blood

Biotechnol Prog. 2020 Nov;36(6):e3056. doi: 10.1002/btpr.3056. Epub 2020 Aug 21.

Abstract

Bacteria must be separated from septic whole blood in preparation for rapid antibiotic susceptibility tests. This work improves upon past work isolating bacteria from whole blood by exploring an important experimental factor: Whole blood dilution. Herein, we use the continuity equation to model red blood cell sedimentation and show that overall spinning time decreases as the blood is diluted. We found that the bacteria can also be captured more efficiently from diluted blood, up to approximately 68 ± 8% recovery (95% confidence interval). However, diluting blood both requires and creates extra fluid that end users must handle; an optimal dilution, which maximizes bacteria recovery and minimizes waste, was found to scale with the square root of the whole blood hematocrit. This work also explores a hypothesis that plasma backflow, which occurs as red cells move radially outward, causes bacterial enrichment in the supernatant plasma with an impact proportional to the plasma backflow velocity. Bacteria experiments carried out with diluted blood demonstrate such bacterial enrichment, but not in the hypothesized manner as enrichment occurred only in undiluted blood samples at physiological hematocrit.

Keywords: antibiotic resistance; modeling; sedimentation; sepsis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacteria / chemistry
  • Bacteria / isolation & purification*
  • Blood Sedimentation*
  • Erythrocytes / microbiology
  • Humans
  • Microbial Sensitivity Tests / methods*

Substances

  • Anti-Bacterial Agents