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Techniques for Isolation of Pure Cultures01:24

Techniques for Isolation of Pure Cultures

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Microorganisms are routinely cultured in the laboratory using various techniques to isolate, grow, and quantify them for further study. These methods rely on inoculating microorganisms into a suitable growth medium under aseptic conditions to prevent contamination. Depending on the objective, inoculation can involve direct transfer or the use of diluted bacterial suspensions as the inoculum.Streak-Plate Method for IsolationThe streak-plate method is a common technique for obtaining pure...
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Updated: Jan 2, 2026

Assaying for Inorganic Polyphosphate in Bacteria
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Assaying for Inorganic Polyphosphate in Bacteria

Published on: January 21, 2019

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A High Throughput Isolation Method for Phosphate-Accumulating Organisms.

Ajeeta Anand1,2, Hideki Aoyagi3

  • 1Institute of Life Sciences and Bioengineering, Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8572, Japan.

Scientific Reports
|December 4, 2019
PubMed
Summary

A novel method isolates elite phosphate-accumulating organisms (PAOs) from probiotics to combat hyperphosphatemia, a complication of chronic kidney disease. This technique enhances PAO screening and offers a healthier alternative to traditional treatments.

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Area of Science:

  • Microbiology
  • Biotechnology
  • Nephrology

Background:

  • Hyperphosphatemia is a common complication of chronic kidney disease.
  • Current treatments like phosphate binders and dialysis have limitations, including side effects and high costs.
  • Probiotics containing phosphate-accumulating organisms (PAOs) offer a potential alternative for managing hyperphosphatemia.

Purpose of the Study:

  • To develop an efficient and reliable method for isolating elite PAOs from probiotic strains.
  • To overcome limitations in current PAO screening and isolation, such as inefficient enrichment and contamination.
  • To identify and quantify the phosphate accumulation capabilities of isolated PAOs.

Main Methods:

  • A novel strategy involving enrichment for phosphate uptake, incubation in low-pH phosphate-free media, and Percoll density gradient centrifugation was employed.
  • Elite PAOs were purified and isolated using toluidine blue O-supplemented agar-based media.
  • Phosphate accumulation capabilities were semi-quantitatively scored and correlated with accumulated phosphate values.

Main Results:

  • The developed technique successfully isolated elite PAOs from Lactobacillus casei and Bifidobacterium adolescentis.
  • Isolated PAOs from L. casei and B. adolescentis demonstrated significantly increased phosphate accumulation (0.81 and 0.70 mg-phosphate/mg-dry cell, respectively).
  • The semi-quantitative scoring method correlated well with the measured phosphate accumulation levels.

Conclusions:

  • The novel isolation method effectively identifies and quantifies elite PAOs with high phosphate accumulation potential.
  • This technique provides a promising approach for developing probiotic-based strategies to prevent or manage hyperphosphatemia.
  • The isolated elite PAOs could serve as a safer and healthier alternative for early-stage hyperphosphatemia management.