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Related Concept Videos

Methods to Assess Microbial Populations01:30

Methods to Assess Microbial Populations

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Assessing microbial populations is crucial for understanding microbial roles in health, ecology, and industry. Various complementary techniques—both culture-based and molecular—enable detailed analysis of microbial abundance, diversity, and function.Viable Plate CountThe viable plate count is a traditional culture-based method used to estimate the number of living microbes in a sample. After serial dilution, the sample is spread onto nutrient agar plates. Each viable cell forms a...
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Methods to Assess Microbial Communities01:19

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Microbial communities, comprising bacteria, archaea, and eukaryotic microorganisms, inhabit diverse ecosystems and play crucial roles in environmental and biological processes. Their diversity is defined by three main parameters: species richness (the number of distinct species), species abundance (the relative quantity of each species), and species evenness (how uniformly individual species are distributed in various locations). These factors together shape the structure and ecological balance...
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The human microbiota begins developing at birth and undergoes continual change as we age. Infancy marks a critical period of microbial sensitivity, offering a “window of opportunity” during which beneficial microbes help mature the immune system. By age three, children typically develop a more stable and diverse microbial community. Newborns acquire microbes from their immediate environment; vaginal delivery favors maternal vaginal microbes, while cesarean births favor microbes from...
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The Oral Microbiota01:27

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The oral microbiome includes a complex ecosystem comprising over 700 microbial species, identified through genomic sequencing and culture-based analyses to date. This community includes a core microbiome, found universally among individuals, and a variable component influenced by environmental factors such as diet, lifestyle, and host genetics. Site-specific conditions, including oxygen gradients, pH levels, and nutrient availability, determine the spatial distribution of these microorganisms...
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Development of the Oral Microbiota01:28

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The establishment of the oral microbiome begins before birth, challenging the long-held belief that the fetal oral cavity is sterile. The presence of oral microbes such as Streptococcus and Fusobacterium in amniotic fluid suggests that microbial exposure may occur in utero, potentially through translocation from the maternal oral or gastrointestinal tract. This early colonization primes the neonatal immune system and sets the stage for subsequent microbial succession. Maternal health,...
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Updated: May 6, 2026

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children
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Standardizing oral microbiome sampling for qPCR: methodological and exploratory insights into nutritional status.

Karina Mendes1, Ana T P C Gomes1, Cristina Maria Mendes Resende2

  • 1Center for Interdisciplinary Research in Health (CIIS), Faculty of Dental Medicine (FMD), Universidade Católica Portuguesa, Estrada da Circunvalação, 3504-505, Viseu, Portugal.

Scientific Reports
|March 15, 2026
PubMed
Summary

Unstimulated saliva is the most consistent method for quantifying oral bacteria using quantitative polymerase chain reaction (qPCR). This reproducible, non-invasive approach is ideal for microbiome studies in adolescent health and nutritional research.

Keywords:
BacillotaBacteroidotaOral microbiomeQPCRSalivaSampling methodology

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Area of Science:

  • Microbiology
  • Human Physiology
  • Molecular Biology

Background:

  • Accurate oral microbiota quantification requires standardized sample collection.
  • Quantitative polymerase chain reaction (qPCR) is a key method for bacterial enumeration.

Purpose of the Study:

  • To evaluate and identify the most consistent oral sample collection method for bacterial quantification in adolescents using qPCR.
  • To assess the biological applicability of the optimal method by comparing bacterial profiles and body composition in adolescents.

Main Methods:

  • Collected unstimulated saliva, cheek swabs, and biofilm samples from adolescents.
  • Utilized qPCR to quantify total bacteria (16S rRNA gene), Bacillota, and Bacteroidota phyla.
  • Performed exploratory analysis correlating bacterial abundance with body composition parameters.

Main Results:

  • Unstimulated saliva yielded the lowest variability in bacterial quantification compared to cheek swabs and biofilm (p < 0.05).
  • Moderate correlations were found between saliva and biofilm, with weak associations between saliva and cheek swabs.
  • While bacterial counts trended higher in overweight/obese adolescents, differences were not statistically significant.

Conclusions:

  • Unstimulated saliva is a validated, reproducible, non-invasive, and cost-effective biofluid for oral microbiota quantification by qPCR.
  • The findings support saliva's utility for large-scale, translational, or point-of-care microbiome assessments in nutritional and metabolic health studies.
  • Group-specific associations between oral bacteria and body composition highlight saliva's potential in health research.