Association Between Periodontal Status and the Prevalence of Entamoeba Gingivalis and Trichomonas Tenax

May 4, 2026 updated by: Bilge MERACI YILDIRAN, Abant Izzet Baysal University

Prevalence of Entamoeba Gingivalis and Trichomonas Tenax in Saliva and Subgingival Plaque Samples Among Periodontology Patients: A Prospective Observational Study

This prospective observational study aims to investigate the prevalence and clinical significance of two oral protozoa, Entamoeba gingivalis and Trichomonas tenax, among patients attending the Bolu Abant İzzet Baysal University Faculty of Dentistry, Department of Periodontology. While the oral microbiome typically maintains a delicate balance, disruptions in this ecosystem are thought to trigger periodontal diseases. Recent evidence suggests that these parasites may contribute to increased inflammation and tissue destruction, potentially playing a role in the etiology of gingivitis and periodontitis.

The study will include 120 participants aged 18 and older who meet the inclusion criteria. Following the collection of demographic data and oral hygiene habits via a questionnaire, a single calibrated examiner will perform comprehensive clinical periodontal examinations. Measurements will include plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing pocket depth (PPD), clinical attachment loss (CAL), and gingival recession (GR) based on the 2017 World Workshop Classification of Periodontal and Peri-Implant Diseases and Conditions.

To detect the presence of parasites, unstimulated whole saliva samples and subgingival plaque samples from the deepest periodontal pockets will be collected from each participant. These samples will be analyzed immediately at the Parasitology Laboratory using light microscopy (10X and 40X magnification) to identify live trophozoites. By evaluating the relationship between parasite prevalence and periodontal status, this research aims to contribute to the limited literature on oral protozoa in Turkey and increase clinical awareness regarding their impact on oral health.

Study Overview

Detailed Description

The primary objective of this single-center, cross-sectional, prospective study is to evaluate the frequency of Entamoeba gingivalis and Trichomonas tenax in patients with different periodontal health statuses. Despite being the first protozoa identified in the oral cavity, the role of these parasites in the pathogenesis of periodontal destruction remains a critical area of investigation. This study seeks to correlate the presence of these protozoa with clinical periodontal parameters and patient-related factors.

  1. Patient Selection and Recruitment:

    A total of 120 patients who apply to the Bolu Abant İzzet Baysal University Faculty of Dentistry, Department of Periodontology, will be screened. Following written informed consent, a detailed questionnaire will be administered to collect data on:

    Demographics (Age, sex, education, and economic status)

    Medical history (Systemic diseases, smoking, and pregnancy status)

    Oral hygiene habits and recent use of antibiotics or antiseptic mouthwashes.

  2. Clinical Periodontal Examination:

    Periodontal diagnosis will be performed according to the "2017 World Workshop Classification of Periodontal and Peri-Implant Diseases and Conditions." All clinical measurements will be conducted by a single calibrated examiner using a Williams periodontal probe. The following parameters will be recorded:

    Plaque index (PI): Evaluated at 4 sites per tooth (0-3 scale).

    Gingival index (GI): Assessment of gingival inflammation (0-3 scale).

    Bleeding on probing (BOP): Presence/absence of bleeding within 10 seconds of probing, expressed as a percentage.

    Probing pocket depth (PPD): Distance from the gingival margin to the base of the pocket (6 sites per tooth).

    Clinical attachment loss (CAL): Distance from the cemento-enamel junction to the base of the pocket.

    Gingival recession: Distance from the cemento-enamel junction to the gingival margin.

  3. Sample Collection and Laboratory Analysis:

    Two types of biological samples will be collected from each participant:

    Subgingival plaque: After isolating the area with cotton rolls and removing supragingival plaque, samples will be collected from the 4 deepest periodontal pockets using sterile curettes and transferred into tubes containing 0.9% NaCl.

    Unstimulated saliva: Approximately 2 mL of whole saliva will be collected after at least 30 minutes of no food intake or mouth rinsing.

    Samples will be transported immediately to the Parasitology Laboratory at Bolu Abant İzzet Baysal University Faculty of Medicine. Three slide preparations per sample will be examined under a light microscope (10X and 40X magnification) to identify the presence of live trophozoites.

  4. Sample Size and Power Analysis:

The sample size of 120 was determined based on similar prevalence studies in the literature.

Study Type

Observational

Enrollment (Estimated)

120

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Locations

    • Gölköy
      • Bolu, Gölköy, Turkey (Türkiye), 14030
        • Recruiting
        • Bolu Abant İzzet Baysal University, Faculty of Dentistry, Department of Periodontology
        • Contact:

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

  • Adult
  • Older Adult

Accepts Healthy Volunteers

Yes

Sampling Method

Non-Probability Sample

Study Population

Patients aged 18 and older who applied to the Bolu Abant İzzet Baysal University Faculty of Dentistry, Department of Periodontology, and met the inclusion criteria.

Description

Inclusion Criteria:

  • Participants aged 18 years or older.
  • Having at least 10 natural teeth.
  • Willingness to undergo periodontal examination and providing written informed consent.
  • Includes both periodontally healthy individuals and patients with gingivitis or periodontitis.

Exclusion Criteria:

  • Use of systemic antibiotics within the last 3 months.
  • Use of chlorhexidine or similar antiseptic mouthwashes within the last month.
  • Having received any periodontal treatment within the last 3 months.
  • Receiving immunosuppressive therapy or having a serious systemic immune deficiency.
  • Pregnancy or breastfeeding.

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

Cohorts and Interventions

Group / Cohort
Intervention / Treatment
Periodontally Healthy
Systemically healthy individuals with no clinical signs of gingival inflammation (BOP < 10%), no clinical attachment loss, and no probing depths > 3 mm.
Collection of unstimulated saliva and subgingival plaque samples followed by immediate light microscopy examination (10X and 40X magnification) to detect the presence and frequency of Entamoeba gingivalis and Trichomonas tenax trophozoites.
Comprehensive periodontal examination including plaque index, gingival index, bleeding on probing, probing pocket depth, and clinical attachment loss measurements using a Williams periodontal probe according to the 2017 World Workshop Classification.
Gingivitis
Patients diagnosed with dental plaque-induced gingivitis characterized by bleeding on probing (BOP ≥ 10%) and no clinical attachment loss or radiographic bone loss, according to the 2017 World Workshop classification.
Collection of unstimulated saliva and subgingival plaque samples followed by immediate light microscopy examination (10X and 40X magnification) to detect the presence and frequency of Entamoeba gingivalis and Trichomonas tenax trophozoites.
Comprehensive periodontal examination including plaque index, gingival index, bleeding on probing, probing pocket depth, and clinical attachment loss measurements using a Williams periodontal probe according to the 2017 World Workshop Classification.
Periodontitis
Patients diagnosed with periodontitis (stages I-IV) characterized by interdental clinical attachment loss, probing depths ≥ 4 mm, and radiographic evidence of alveolar bone loss.
Collection of unstimulated saliva and subgingival plaque samples followed by immediate light microscopy examination (10X and 40X magnification) to detect the presence and frequency of Entamoeba gingivalis and Trichomonas tenax trophozoites.
Comprehensive periodontal examination including plaque index, gingival index, bleeding on probing, probing pocket depth, and clinical attachment loss measurements using a Williams periodontal probe according to the 2017 World Workshop Classification.

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Prevalence of Entamoeba gingivalis and Trichomonas tenax
Time Frame: Baseline (at the time of clinical examination and sample collection, typically within 1 hour).
The presence or absence of live trophozoites of Entamoeba gingivalis and Trichomonas tenax will be determined through light microscopic examination (10X and 40X magnification) of saliva and subgingival plaque samples. The outcome will be reported as the percentage (%) of participants testing positive for each parasite within the different periodontal health groups.
Baseline (at the time of clinical examination and sample collection, typically within 1 hour).

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

December 22, 2025

Primary Completion (Estimated)

July 1, 2026

Study Completion (Estimated)

September 1, 2026

Study Registration Dates

First Submitted

April 20, 2026

First Submitted That Met QC Criteria

April 20, 2026

First Posted (Actual)

April 24, 2026

Study Record Updates

Last Update Posted (Actual)

May 8, 2026

Last Update Submitted That Met QC Criteria

May 4, 2026

Last Verified

May 1, 2026

More Information

Terms related to this study

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

NO

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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