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Microbial Growth Measurement: Direct Methods

Direct methods for measuring microbial populations in a culture are essential tools in microbiology, providing quantitative data for various applications. Among these, microscopic counts, plate counts, and serial dilution are widely used techniques, each with unique principles and applications.Microscopic CountsMicroscopic counting involves the use of a Petroff-Hausser chamber, a specialized microscope slide with a grid and defined depth. By observing a liquid culture under a microscope,...
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A novel method to quantify viable Enterococcus faecium during feed manufacturing.

Ying Zhang1, Songjun Jiao1, Xuetao Liu1

  • 1The State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

Poultry Science
|June 30, 2026
PubMed
Summary
This summary is machine-generated.

A new propidium monoazide (PMA)-qPCR method accurately quantifies viable Enterococcus faecium (E. faecium) in feed. Pre-encapsulation significantly enhances E. faecium survival during feed manufacturing, improving probiotic efficacy.

Keywords:
Coating technologyEnterococcus faeciumFeed manufacturingPropidium monoazide-qPCRViable cell

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Area of Science:

  • Microbiology
  • Animal Nutrition
  • Food Science

Background:

  • Enterococcus faecium (E. faecium) viability is crucial for probiotic efficacy but is compromised during feed manufacturing.
  • Existing methods lack accuracy in quantifying live E. faecium in feed matrices.
  • Feed processing conditions, such as heat and mechanical stress, reduce bacterial survival.

Purpose of the Study:

  • To develop and validate a propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) method for quantifying viable E. faecium in feed.
  • To evaluate the efficacy of different coating technologies in protecting E. faecium during feed manufacturing.
  • To assess the impact of feed processing parameters on the survival of coated and uncoated E. faecium.

Main Methods:

  • Developed a PMA-qPCR assay, optimizing propidium monoazide concentration and validating primer efficiency and specificity.
  • Assessed cell release from coated E. faecium using artificial intestinal fluid with trypsin.
  • Investigated the survival of uncoated, post-encapsulated, and pre-encapsulated E. faecium under varying feed manufacturing conditions (temperature, conditioning, pelleting) using the validated PMA-qPCR method.

Main Results:

  • The established PMA-qPCR method demonstrated high sensitivity (E = 101.1%; R² = 0.991) and accuracy in detecting live and dead E. faecium.
  • Survival of E. faecium decreased significantly with increasing conditioning temperatures (72.4% at 65°C to 12.2% at 85°C).
  • Pre-encapsulated E. faecium exhibited significantly higher survival rates compared to uncoated and post-encapsulated forms throughout all feed manufacturing steps and temperatures.

Conclusions:

  • The PMA-qPCR method is a valuable tool for accurately quantifying viable E. faecium in feed.
  • Pre-encapsulation technology offers superior protection for E. faecium against heat and mechanical stress during feed processing.
  • Enhanced E. faecium survival through pre-encapsulation can improve probiotic efficacy in animal feed.