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Related Experiment Video

Updated: Jul 2, 2026

Discovery of New Intracellular Pathogens by Amoebal Coculture and Amoebal Enrichment Approaches
09:04

Discovery of New Intracellular Pathogens by Amoebal Coculture and Amoebal Enrichment Approaches

Published on: October 27, 2013

Novel Parachlamydia acanthamoebae quantification method based on coculture with amoebae.

Junji Matsuo1, Yasuhiro Hayashi, Shinji Nakamura

  • 1Department of Medical Laboratory Sciences, Faculty of Health Sciences, Hokkaido University, Sapporo, Hokkaido, Japan.

Applied and Environmental Microbiology
|September 2, 2008
PubMed
Summary

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A new method quantifies viable Parachlamydia acanthamoebae (P. acanthamoebae) by co-culturing with amoebae. This technique accurately measures bacterial growth in host cells, aiding pathogen research.

Area of Science:

  • Microbiology
  • Bacteriology
  • Infectious Diseases

Background:

  • Parachlamydia acanthamoebae is an obligate intracellular bacterium infecting amoebae.
  • It is a potential human pathogen, but viable bacterial quantification methods are lacking.

Purpose of the Study:

  • To develop and validate a novel method for quantifying viable Parachlamydia acanthamoebae.

Main Methods:

  • Co-culturing P. acanthamoebae with Acanthamoeba spp. or mammalian cell lines.
  • Quantification using DAPI staining and fluorescent in situ hybridization to determine amoeba-infectious dose (AID).
  • Calculation of amoeba-infectious units (AIU) based on logistic sigmoid dilution curves.

Main Results:

  • The novel method accurately monitored P. acanthamoebae growth in amoebae, reaching a 1,000-fold increase within 4 days.

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Published on: October 27, 2013

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  • Amoeba viability and morphology remained stable during co-culture.
  • No significant bacterial increase was observed in mammalian cell lines, though aberrant structures suggested persistent infection.

    • Conclusions:

    • The developed co-culture method effectively quantifies viable P. acanthamoebae (AIU).
    • This technique is valuable for studying chlamydiae in environmental and pathogenic contexts.
    • Further research may elucidate persistent infection mechanisms in mammalian cells.