Detection of trichomonads in patients with lung cancer and transcription analysis on the response of human pulmonary epithelial cells to Trichomonas tenax invasion.

INTRODUCTION: Lung cancer is one of the most prevalent malignancies worldwide. Substantial research has illuminated the intricate interplay between microorganisms and human health, revealing their role in disease regulation. Trichomonads is a flagellated protozoan in the human cavity and have been previously identified as a pathogen associated with pneumonia, contributing to tissue chronic inflammation and carcinogenesis. METHODS: Nested polymerase chain reaction methods were employed to scrutinize the prevalence of trichomonads in the bronchovesicular fluid of patients diagnosed with lung cancer. Subsequently, the influence of Trichomonas tenax invasion on lung cancer cells was elucidated through proliferation assays, migration assays, and transcription analysis. RESULTS: Bronchoalveolar fluid samples from lung cancer patients yielded positive nested PCR results for eight out of twenty-seven samples. Seven of these samples were identified as Trichomonas tenax, while one was identified as Tetratrichomonas spp. Our findings revealed a significant upregulation of pathways associated with carcinogenesis, including cellular proliferation, migration, and drug resistance, in response to T. tenax invasion. CONCLUSIONS: This study underscores the importance of recognizing the presence of trichomonads and the influence of T. tenax invasion on host responses to respiratory diseases. The identified pathways implicated in cancer development may pave the way for developing targeted treatment strategies for pulmonary diseases. These findings hold promise for informing and improving the precision of therapeutic interventions in the context of pulmonary ailments.

Risk factors and approaches for detection of Trichomonas tenax, the silent culprit in periodontal disease: A narrative review.

Introduction: Periodontal disease is the inflammation of the periodontium tissues surrounding the teeth, potentially leading to loss of tooth attachment. In individuals with periodontal disease, the presence of Trichomonas tenax, a parasitic protozoan of the oral cavity has been observed and its frequency tends to rise as the disease progresses. Methods: A literature search was conducted in the online databases of PubMed, Google Scholar, Web of Science, and Scopus using the combination of keywords: "Trichomonas tenax" AND "periodontal disease" OR "gum disease", OR "oral disease" OR "periodontitis". A total of 9 articles satisfied the inclusion criteria and were included in this study. Results: This review highlights the incidence of T. tenax with periodontal diseases, the risk factors that contribute to the infection of T. tenax and available detection methods for the identification of the protozoan. Conclusion: The inhabitation of the oral cavity by T. tenax prospers with the severity of periodontal diseases. Extensive research should be conducted to fully understand the potential pathogenic role and damaging effect of T. tenax in the oral cavity.

Clinical implications of trichomonads detected in bronchoalveolar fluid by metagenomic next-generation sequencing: a multicenter retrospective study.

Background: Pulmonary trichomoniasis is considered a neglected disease due to failures in recognizing it, stemming from insensitive microbial methods and a lack of specific clinical features. This study aims to analyze the clinical implications of trichomonads detected in bronchoalveolar lavage fluid (BALF) by metagenomic next-generation sequencing (mNGS). Methods: This multicenter retrospective study included patients diagnosed with pneumonia, admitted to three tertiary hospitals in China from July 2018 to September 2022, with trichomonads detected in BALF through mNGS. The analysis covered demographics, comorbidities, symptoms, laboratory findings, mNGS results, clinical treatment, and outcomes of these patients. Results: A total of 17 patients were enrolled, comprising 14 males and 3 females. Trichomonas tenax and Trichomonas vaginalis were detected by mNGS in BALF samples of 15 and 2 patients, respectively. Patients were categorized into two groups based on the presence of risk factors for trichomonad infection, including immunocompromised conditions, uncontrolled diabetes mellitus, oral/periodontal diseases, and aspiration. Among 11 patients with risk factors (Case 1-11), 4 received nitromidazoles as part of comprehensive treatment, achieving a 100% treatment success rate. The remaining 7 patients, who did not receive nitromidazoles, had only one achieving relief after broad-spectrum antimicrobial therapy, resulting in a 14.3% treatment success rate. For the 6 patients without any risk factors for trichomonad infection (Case 12-17), none received nitromidazoles during hospitalization. However, 4 out of these 6 patients (66.7%) eventually recovered. Conclusion: mNGS proves to be an efficient tool for detecting trichomonads in BALF samples. Comprehensive analysis of clinical features and laboratory indicators is essential to distinguish between infection and colonization of trichomonads. Pulmonary trichomoniasis should not be overlooked when trichomonads are detected in BALF from patients with risk factors.

Prevalence of Entamoeba gingivalis and Trichomonas tenax among patients with periodontal disease attending Dental Clinic, University College Hospital, Ibadan.

Background: Periodontal disease is often caused by bacterial plaque. However, there are indications that some infective agents, including parasites, may play important roles in the pathogenesis of the disease. Aim: This study aimed at assessing the prevalence of gingivitis and periodontitis, as well as the prevalence of Entamoeba gingivalis and Trichomonas tenax, in the oral biofilm of individuals with periodontal diseases using polymerase chain reaction. Materials and Methods: One hundred and six respondents with periodontal disease participated in the study. All study participants had a full-mouth examination, and dental plaque samples were collected with a sterile curette. Samples were transported to the laboratory in a cold chain and stored frozen till analyzed. DNA was extracted from the samples and amplified using EGO and ENTAM primers for Entamoeba and TGBK primers for T. tenax. Results: The mean age of respondents was 45 ± 16.3 years, with none using tobacco. The prevalence of gingivitis and periodontitis obtained from this study was 84.9% and 15.1%. The prevalence obtained for Entamoeba and T. tenax was 0.9% each; however, no participant had both parasites. The positive samples were from male participants with moderate gingivitis. Conclusion: Gingivitis was more prevalent than periodontitis, though with a high preponderance in females. E. gingivalis and T. tenax may not be of etiologic importance in periodontal disease as they occurred sparsely in the studied population.

Molecular Epidemiology and Associated Risk Factors of Parasites in Oral Cavity of Children with Malignancies in Western Iran.

Background: This survey was designed to study the molecular epidemiology and risk factors of Entamoeba gingivalis and Trichomonas tenax in children with underlying malignancies and those on chemotherapy in Lorestan province, West of Iran. Methods: The present cross-sectional descriptive study was performed on children who suffering from different types of malignancies or receiving treatment by chemotherapy referring to oncology section of hospitals of Lorestan Province, Iran during May 2021 to April 2022. The frequency of oral cavity protozoa was investigated using microscopic and conventional polymerase chain reaction (PCR). Results: E. gingivalis and T. tenax parasites were found in 23 (25.5%) by microscopic method and 28 (31.1%) using PCR in children with malignancy. Among positive samples, 20 (71.4%) were infected with E. gingivalis; whereas 8 (28.6%) of the participants were positive for T. tenax. In the multivariate model, living in rural regions (OR= 3.437; 95% CI= 1.22-9.63; p=0.019) and using mouthwash (OR= 0.082; 95% CI= 0.018-0.37; p<0.001) were significantly related with the frequency of oral cavity parasites. Conclusion: Our results showed the high frequency of oral cavity parasites in children who suffering malignancies or receiving treatment by chemotherapy in Lorestan province, Iran. The awareness of the main risk factors for oral cavity parasites particularly using mouthwash is necessary in improving public and oral health strategies in children with cancer. Consequently, oncologist and dental practitioners must be aware to identify and manage oral health concerns in in children who suffering from different types of malignancies to prevent the oral diseases and infections.

Molecular epidemiology and associated risk factors of oral cavity parasites in hemodialysis patients in western Iran.

Renal disorders are able to make patients more susceptible to infections, including those caused oral cavity infections. Today, it has been proven that during end stage renal failure, hemodialysis results in acquired immune response defects through failings of humoral and cell-mediated immune. This survey was intended to assess the molecular epidemiology and associated risk factors of Entamoeba gingivalis and Trichomonas tenax as the main oral cavity protozoa in hemodialysis patients in Lorestan Province, western Iran. The investigation was performed on 73 hemodialysis patients referring to hemodialysis centers of Lorestan Province, Iran during May 2021 to February 2022. The frequency of oral cavity protozoa was investigated using microscopic and conventional polymerase chain reaction (PCR). A provided questionnaire with a number of demographical information and related risk factors was completed for each patient. The results showed that E. gingivalis and T. tenax parasites were found in 18 (24.6%) and 20 (27.4%) of the hemodialysis participants by microscopic and PCR test, respectively. Among samples, 13 (17.8%) of the hemodialysis participants were infected with E. gingivalis; whereas 7 (9.6%) of the participants were positive for T. tenax. No significant association was reported between gender, age, education, flossing, use of mouthwash, and prevalence of oral protozoa in hemodialysis participants. However, a significant correlation was observed among between living in rural regions (OR = 13.16; 95% CI = 2.64-56.81; p = 0.002), brushing teeth (OR = 8.51; 95% CI = 1.71-42.3; p = 0.009) and prevalence of oral protozoa in hemodialysis participants. The findings of these epidemiological study clearly showed the high frequency of oral cavity parasites in hemodialysis patients in Lorestan Province, Western Iran. Awareness of the main risk factors for oral cavity parasites particularly teeth brushing is necessary in refining public and oral health policies in hemodialysis patients. Consequently, dental practitioners, nephrologist, and urologist must be alert of these risk factors to carefully identify and achieve oral health concerns in hemodialysis patients to prevent the oral diseases and infections.

Trichomonas tenax induces barrier defects and modulates the inflammatory cytotoxicity of gingival and pulmonary epithelial cells.

BACKGROUND: Trichomonas tenax is a single-cell flagellated anaerobic organism, commensal in the human oral cavity. Although a previous study indicated that T. tenax could cause cell damage and phagocytose host epithelial cells, its pathological effects on gum cells remain unknown. Furthermore, several case reports have detected T. tenax in several patients with empyema and/or pleural effusion, which may have been aspirated from the oral cavity. However, the cytotoxic effects and immune responses of alveolar cells are unknown. Therefore, we aimed to determine the cytotoxic and immune effects of T. tenax on gums and pulmonary cell lines. The cytopathic effect and lactate dehydrogenase (LDH) cytotoxicity assays were used to determine the level of cell damage in gum and lung epithelial cells. Western blot was used to determine the disruption of cell junctions. Finally, epithelial cell cytokines were measured using ELISA to elucidate the immune response to T. tenax. RESULTS: We found that T. tenax induced a cytotoxic effect on gum epithelial cells by disrupting cell junctions; however, it hardly triggered cellular damage in alveolar A549 cells and mucoepidermoid NCI-H292 cells. Furthermore, T. tenax induced the production of IL-6 at a low multiplicity of infection (MOI) in gum, A549, and NCI-H292 cells. CONCLUSIONS: Our results suggest that T. tenax can trigger gingival cell cytotoxicity, disrupt cell junctions, and induce IL-6 production in gingival and pulmonary cell lines.

Title: Trichomonas tenax induit des défauts de barrière et module la cytotoxicité inflammatoire des cellules épithéliales gingivales et pulmonaires. Abstract: Contexte : Trichomonas tenax est un organisme anaérobie unicellulaire flagellé, commensal dans la cavité buccale humaine. Bien qu'une étude précédente ait indiqué que T. tenax pouvait endommager les cellules et phagocyter les cellules épithéliales de l'hôte, ses effets pathogènes sur les cellules gingivales restent inconnus. En outre, plusieurs rapports ont détecté T. tenax chez des patients présentant un empyème et/ou un épanchement pleural, qui peut avoir été aspiré de la cavité buccale. Cependant, les effets cytotoxiques et les réponses immunitaires des cellules alvéolaires sont inconnus. Par conséquent, nous avons cherché à déterminer les effets cytotoxiques et immunitaires de T. tenax sur des lignées cellulaires de gencives et de poumons. Les tests d'effet cytopathique et de cytotoxicité de la lactate déshydrogénase (LDH) ont été utilisés pour déterminer le niveau de dommages cellulaires dans les cellules. Le Western Blot a été utilisé pour déterminer la perturbation des jonctions cellulaires. Enfin, les cytokines des cellules épithéliales ont été mesurées par ELISA pour élucider la réponse immunitaire à T. tenax. Résultats : Nous avons constaté que T. tenax induisait un effet cytotoxique sur les cellules épithéliales gingivales en perturbant les jonctions cellulaires. Cependant, T. tenax a déclenché des dommages cellulaires seulement mineurs dans les cellules alvéolaires A549 et les cellules mucoépidermoïdes NCI-H292. De plus, T. tenax a induit la production d'IL-6 à une faible multiplicité d'infection (MOI) dans les cellules de gencives, A549 et NCI-H292. Conclusions : Nos résultats suggèrent que T. tenax peut déclencher la cytotoxicité des cellules gingivales, perturber les jonctions cellulaires et induire la production d'IL-6 dans les lignées cellulaires gingivales et pulmonaires.

Entamoeba gingivalis and Trichomonas tenax: Protozoa parasites living in the mouth.

OBJECTIVE: This review article aims to summarize the existing data on the history, biology and potential pathogenicity of Entamoeba gingivalis and Trichomonas tenax in periodontal disease, as well as the available techniques for laboratory diagnosis. DESIGN: A detailed review of scientific literature available up to October 1, 2022 in three databases (PubMed, Scopus and Web of Science) was performed relevant to biology, biochemistry, epidemiology, and experimental studies on infection by E. gingivalis and T. tenax, as well as laboratory techniques for the diagnosis of both protozoa in periodontal diseases. RESULTS: Accumulated evidence over the decades indicates that the protozoa E. gingivalis and T. tenax are able to interact with host cells and induce inflammation in the periodontal tissue by promoting the expression of pro-inflammatory molecules and the recruitment of neutrophils, contributing to the periodontal disease process. Among the available techniques for the laboratory diagnosis, culture and molecular assays seems to be the best tools for detection of both protozoan parasites. CONCLUSIONS: E. gingivalis and T. tenax are potentially pathogens that colonize the oral cavity of humans and may cause periodontal disease.

Trichomonas tenax: A Neglected Protozoan Infection in the Oral Cavities of Humans and Dogs-A Scoping Review.

Trichomonas tenax is a flagellated protozoan parasite found in the oral cavities of humans and animals and has been associated with periodontal disease, the most prevalent inflammatory disease affecting them all. Studies have shown that T. tenax can cause damage to mammalian cells and secretes virulent proteins, such as cysteine. It is presently considered zoonotic. Despite the few studies that have been done, the pathogenicity of this oral protozoan is still not fully understood. A database search was performed in July 2022 using PubMed and Google Scholar to retrieve data eligible for this study. PRISMA-ScR guidelines were followed to conduct this scoping review. A total of 321 articles were found with 87 included in this review after applying the exclusion criteria. Due to its increasing prevalence worldwide in both humans and dogs, detecting and elucidating the pathogenicity of this parasite is paramount for effective global control and prevention of periodontal disease. However, there is a paucity in the literature on this neglected zoonotic trichomonad, which is in large contrast to the closely related human pathogen T. vaginalis. Here, we comprehensively review the history, morphology and reproduction, host, prevalence, diagnosis, pathogenicity, control, and prevention of T. tenax. Hopefully, this article will call attention to both medical and veterinary professionals as well as epidemiologists on this most neglected and zoonotic protozoan. More epidemiological and clinical studies need to be conducted on T. tenax to gain a better understanding of its pathogenicity, to increase the chances of developing effective drugs to aid in the control of this oral parasite, and reduce the spread of periodontal disease worldwide.

Molecular prevalence and zoonotic potential of trichomonads from oral cavities in household dogs.

This study investigated the molecular prevalence of oral trichomonads in household dogs. Of the 144 dogs, 21 (14.6%, 21/144) tested positive for oral trichomonads. The prevalence was significantly higher in dogs with severe gingivitis (gingival index 3: 30.0%, 8/26) than that in normal dogs (gingival index 0: 2.7%, 1/37). Therefore, an interaction between oral trichomonads and the development of periodontal disease is suggested. Of the 21 positive samples, 16 isolates were T. brixi, four isolates were T. tenax, and one was Tetratrichomonas sp. Considering T. tenax is recognized as a zoonotic agent, transmission between dogs and humans cannot be neglected.

Molecular detection of oral Trichomonas tenax in periodontal disease patients by polymerase chain reaction -based 18S rRNA gene

@#Trichomonas tenax, an oral flagellated protozoon found in humans, potentially associated with the inflammation of periodontal tissues and decreased immunity that causes the tissue damage and tooth loss from chronic infection. Currently, there is a lack of data regarding the prevalence of T. tenax infection in Thailand. Therefore, this study aimed to measure prevalence of T. tenax in periodontal disease patients by using polymerase chain reaction (PCR) to amplify the 18S ribosomal RNA (18S rRNA) gene and to determine the factors associated with the presence of this protozoan. A cross-sectional descriptive study was conducted among 230 patients with periodontal disease, who visited the oral health center of Suranaree University of Technology Hospital, Thailand from 2021 to 2022. Dental plaque specimens were collected and examined to identify the presence of T. tenax using the PCR-based 18S rRNA gene. The occurrence of factors associated with T. tenax infection was analyzed by the chi-square test and binary logistic regression. The prevalence of T. tenax infection was 13.48% (31/230), in patients, including 96.77% (30/31) and 3.23% (1/31) in periodontitis and gingivitis patients, respectively. The presence of T. tenax was associated with periodontal disease (p<0.001) and the Periodontal Screening and Record (PSR) index (p=0.001). The significant risk factors for T. tenax infection were periodontitis (ORadj=239.89, 95% CI=23.801-2417.746), no-underlying disease (ORadj=0.31, 95% CI=0.099–0.942), and male sex (ORadj=0.25, 95% CI=0.062-0.981). Dentists should be concerned about this oral protozoan in periodontitis patients. Furthermore, epidemiologic studies of T. tenax are still needed to investigate the mechanism of pathogenesis from T. tenax infection.

Oral Protozoa Entamoeba Gingivalis And Trichomonas Tenax Among Periodontitis And Gingivitis Patients In Periodontology Department Dental Section Quetta.

Background: A group of eukaryotic organisms that are either free-living or parasitic and feed on organic matter, such as microorganisms or organic detritus. This group is referred to as a polyphyletic classification. They can be found in a wide variety of patterns and sizes, ranging from an amoeba, which can vary its shape, to a paramecium, which has a fixed shape and a complicated structure. The objective was to find out the occurrence of Entamoeba gingivalis and Trichomonas among Dental Patients visiting the Periodontology Department Dental Section, Sandeman, Provincial Hospital Quetta. Methods: The study design was randomized cross-sectional and conducted in the Department of Periodontology Dental Section, Sandeman Provincial Hospital Quetta, from March 2022 to February 2023. A total of 110 known cases of periodontitis and gingivitis were recruited in this study, and further to laboratory work the bacterial biofilm samples were collected from both gingivitis and periodontitis patients. The dental plaque was placed on individual glass microscope slides and a drop of saline and mixed and covered with a coverslip. The wet smear was examined immediately under 40X, and the objective for the presence and absence of motile amoebae or flagellated protozoa was recorded. . Results: out of 110 known cases of periodontitis and gingivitis 91 (83%) patients were males and 19 (17%) were females among 110 patients, 40 (36%) patients were periodontitis and 70 (64%) patients were gingivitis. Among 40 (36%) periodontitis patients 3(7.5%) were Entamoeba gingivalis and 4(10%) were Trichomonas Tenax while 70 (64%) cases of Gingivitis 20 (28%) of Entamoeba gingivalis and 1(1%) Trichomonas tenax was recorded. No patient had both species of protozoa were seen in this study. Conclusion: The results of this study revealed the frequency of occurrence of E. gingivalis and T. tenax and the state of periodontitis and gingivitis. The prevalence of E. gingivalis was higher than T. tenax. We recommended further research with a higher number of patients and may use more advanced and reliable laboratory techniques such as PCR and electron microscopy.

Prevalencia de Entamoeba gingivalis y Trichomonas tenax en adolescentes de edad escolar / Prevalence of Entamoeba gingivalis and Trichomonas tenax in school-age adolescents

Entamoeba gingivalis y Trichomonas tenaxson protozoos que generalmente se encuentran en la cavidad oral de los humanos. E. gingivalis vive en la superficie de los dientes y las encías, bolsas gingivales cerca de la base de los dientes y rara vez en las criptas de las amígdalas. Por lo general, se transmiten por contacto directo de una persona a otra al besarse, rociarse con gotas o compartir utensilios para comer. Se sabe que hasta el 95% de las personas con poca higiene bucal pueden estar infectadas con esta ameba. Por otro lado, Trichomonas tenax es una pequeña tricomona que generalmente se encuentra en la cavidad oral del 5 al 10% de los humanos. Aunque se consideran comensales anaeróbicos no patógenos, el refugio de este protozoo es más común entre personas con mala higiene bucal o enfermedades. Los informes epidemiológicos demostraron variables tasas de prevalencia de la infección por E. gingivalis entre los niños, y su ocurrencia puede variar de acuerdo a la edad, presencia de gingivitis, periodontitis y condiciones inmunosupresoras. Los niños y adolescentes en edad escolar son extremadamente susceptibles a una amplia gama de infecciones gingivales como bacterias y protozoos. El objetivo de este trabajo es determinar la prevalencia de los protozoarios E. gingivalis y T. tenax en adolescentes con edades comprendidas entre 10 y 19 años en una población de 185 pacientes de la ciudad peruana de Huancayo, Perú. La prevalencia fue mayor en la población femenina (14,05%) que en la población masculina (11,35%), con un mayor porcentaje de prevalencia entre las edades de 17 a 19 años (12,43%), pH salival menor a 6,7 y con enfermedades periodentales (16,22%). Los resultados estadísticos demostraron que la edad (p=0,001), pH salival (p=0,024) y diagnóstico de otras patologías periodentales (p<0,001) fueron estadísticamente significativas(AU)

Entamoeba gingivalis and Trichomonas tenax are protozoa that are usually found in the oral cavity of humans. E. gingivalis lives on the surface of the teeth and gums, gum pockets near the base of the teeth, and rarely in the crypts of the tonsils. They are usually spread by direct person-to-person contact through kissing, dousing, or sharing eating utensils. It is known that up to 95% of people with poor oral hygiene may be infected with this amoeba. On the other hand, Trichomonas tenax is a small trichomona that is usually found in the oral cavity of 5-10% of humans. Although they are considered non-pathogenic anaerobic commensals, the harborage of this protozoan is more common among people with poor oral hygiene or disease. Epidemiological reports have shown variable prevalence rates of E. gingivalis infection among children, and its occurrence may vary according to age, presence of gingivitis, periodontitis, and immunosuppressive conditions. School-age children and adolescents are extremely susceptible to a wide range of gingival infections including bacteria and protozoa. The objective of this work is to determine the prevalence of the protozoa E. gingivalis and T. tenax in adolescents between the ages of 10 and 19 in a population of 185 patients from the Peruvian city of Huancayo, Peru. The prevalence was higher in the female population (14.05%) than in the male population (11.35%), with a higher percentage of prevalence between the ages of 17 to 19 years (12.43%), lower salivary pH to 6.7 and with periodontal diseases (16.22%). The statistical results showed that age (p=0.001), salivary pH (p=0.024) and diagnosis of other periodontal pathologies (p<0.001) were statistically significant(AU)

Pyopneumothorax Caused by Trichomonas tenax and Porphyromonas endodontalis Coinfection in a Patient with Previous Cerebral Infarction: A Case Report and Literature Review.

Background: Even with the advent of NGS and PCR diagnostic tools, cases of chest infections caused by Trichomonas are still very rare. Such pathogens are less likely to be considered by clinicians. These cases frequently involve the pleura and lead to pneumothorax, hydropneumothorax, or pyopneumothorax, making the disease severe. Case Presentation: A 69-year-old man diagnosed with cerebral infarction a year ago sought medical attention for right-sided pyopneumothorax and respiratory failure. The pathogen found in the pleural fluid was highly suspected to be Trichomonas tenax (T. tenax). Pleural fluid mNGS confirmed T. tenax and Porphyromonas endodontalis coinfection. Metronidazole combined with piperacillin tazobactam was administered to counteract infection. Simultaneously, closed chest drainage and thoracoscopic release of pleural adhesions were performed. The patient was cured, discharged from the hospital, and was in good condition after six months of follow-up. Conclusion: When chest infections occur in patients with poor oral hygiene and underlying diseases that may lead to aspiration, the identification of Trichomonas infection should be noted. Early confirmation of the diagnosis often requires mNGS and PCR. Metronidazole is essentially effective against Trichomonas, and medical thoracoscopy can be used to manage pleural conditions if necessary.

Prevalence and risk factors of oral cavity parasites in pregnant women in Western Iran.

Background: We aimed to study the frequency and risk factors of oral cavity protozoa (Trichomonas tenax and Entamoeba gingivalis) in pregnant women in Lorestan province, western Iran. Methods: The current cross-sectional descriptive work was performed on 200 pregnant women referring to health centers of Lorestan Province, Iran during August 2021 to February 2022. Two specimens were obtained from each women by means of sterile swabs from saliva and dental plaques. The frequency of oral cavity protozoa was investigated using microscopic and conventional polymerase chain reaction (PCR). Results: E. gingivalis and T. tenax parasites were found in 41 (20.5%) and 46 (23%) of the pregnant women by microscopic and PCR test, respectively. Among positive samples, 29 (63.1%) of the pregnant women were infected with E. gingivalis; whereas 19 (36.9%) of the participants were positive for T. tenax. In the multivariate model, living in rural regions (p < 0.001) and brushing teeth (p = 0.021) were considerably linked with the prevalence of oral cavity parasites. Conclusion: The current study evidently exhibited the high frequency of oral cavity parasites (E. gingivalis and T. tenax) in pregnant women in Lorestan province, Western Iran. Awareness of the main risk factors for oral cavity parasites especially teeth brushing is necessary in refining public and oral health approaches in pregnant women. Thus, dental practitioners and gynecologists must be alert of these risk factors to carefully identify and manage oral health concerns in pregnant women to prevent the oral diseases and infections.

Multiplex PCR assay targeting Trichomonas vaginalis: need for biological evaluation and interpretation.

In a retrospective study, we used sequencing to investigate Trichomonas vaginalis-positive specimens (genital, rectal and pharyngeal) with the Allplex™ STI Essential or the Anyplex™-II-STI-7 assays. Our results confirm that majority of T. vaginalis-positive genital and pharyngeal specimens contained T. vaginalis DNA and actually T. tenax DNA, respectively.

Assessment of Entamoeba gingivalis and Trichomonas tenax in Patients with Chronic Diseases and its Correlation with Some Risk Factors.

Trichomonas tenax (T. tenax) and Entamoeba gingivalis (E. gingivalis) are two oral protozoan parasites that are universal and found in patients with poor oral hygiene, as well as chronic and periodontal diseases. This study was conducted to assess the efficacy of some parameters, such as age, gender, education, residency, smoking, and dental cleaning routines, on the incidence of these parasites in the oral cavity of the patients participating in this study. A total of 230 individuals with chronic diseases, 97 of whom were patients (44 females and 53 males) participated in the present study. Dental plaque and saliva samples were collected from each patient and examined under light microscopy with Giemsa staining. Out of 230 dental plaque samples, 60 (26.08%) samples were positive for E. gingivalis, while 37 (16.08%) cases were positive for T. tenax. Diabetic patients showed high E. gingivalis percentage (n=12; 20%) (P=0.000*). However, thyroid disorders showed a low percentage. Hypertensive patients showed a high percentage for T. tenax (n=6; 16.21%) (P=0.000*), while tuberculosis patients showed the lowest percentage. Patients with hypertension, smoking, heart disease, and diabetes showed statistical significance for the presence of these parasites. Equally, patients older than 30 years have shown a higher rate of infection. According to the findings, E. gingivalis was detected in 58.33% and 41.66% of the urban and rural populations, respectively. Furthermore, T. tenax was detected in 81.08% and 18.91% of the urban and rural populations, respectively. Parasitic contagions were comparatively popular in patients with chronic and periodontal illness.

Parasites in Periodontal Health and Disease: A Systematic Review and Meta-analysis.

BACKGROUND: Periodontitis is an inflammatory disease triggered by the infection of the periodontal sulcus by microbes. Together with the abundant eubacterial microbiota, at least two parasites have often been identified: the amoeba Entamoeba gingivalis and the flagellate Trichomonas tenax. The role of these protists in the pathophysiology of periodontal disease remains to be deciphered. A high diversity in their measured prevalence, mainly due to methodological concerns, prevents further analysis of the aetiological link between these parasites and periodontitis. METHODS: To determine E. gingivalis and T. tenax prevalence in periodontal pockets as compared to healthy sulci, we have conducted a systematic review, searching 3 remote databases (Pubmed, LILACS, and Google Scholar), restricting to papers in which the diagnostic of the parasite was made using molecular methods. A total of 5 studies for E. gingivalis and 2 studies for T. tenax were included for the meta-analysis. RESULTS: In the periodontal pockets, the prevalence of parasites is 76.9% (95%-CI: 71.5-81.7%) for E. gingivalis and 38.6% (95%-CI: 27.2-50.0%) for T. tenax . Both parasites are more abundant in periodontal pockets as compared to healthy sulci, with a risk ratio of 3.96 (95%-CI: 1.57-9.98) for E. gingivalis and 21.82 (95%-CI: 6.71-70.96) for T. tenax . The two subtypes of E. gingivalis exhibited the same risk ratio: 3.30 (95%-CI: 1.27-8.55) for ST1 and 3.30 (95%-CI: 0.42-26.03) for ST2, but ST1 was more prevalent (70.6%, 95%-CI: 65.0-76.2%) than ST2 (43.9%, 95%-CI: 35.5-52.4%) in periodontal pockets. CONCLUSION: Altogether, the data show that parasites are more prevalent in the diseased than in the healthy. However, the differences in prevalence between species and subtypes call for more studies to be able to conclude about their individual contributions in the pathophysiology of periodontal diseases. The heterogeneity in prevalence estimation should be investigated further, in particular to make out biological from methodological heterogeneity.

A Loop-Mediated Isothermal Amplification (LAMP) Assay Specific to Trichomonas tenax Is Suitable for Use at Point-of-Care.

Trichomonas tenax is a flagellated protozoan that inhabits the human and canine oral cavity in patients with poor oral hygiene and periodontal disease. The loop-mediated isothermal amplification (LAMP) assay could provide clinicians with a quick, cheap and reliable diagnostic test used for the detection of T. tenax in various settings. In this study, we aimed to develop a LAMP assay that can detect T. tenax with high sensitivity and specificity. A set of LAMP primers were specifically designed to detect the ITS and 5.8S rRNA gene of T. tenax. The newly developed LAMP assay was 1000 times more sensitive than conventional PCR. The limit of detection of the LAMP assay was 10 fg of genomic DNA, or 0.2-1 cell. Moreover, the LAMP assay was specific, resulting in no cross-reaction even with a closely related protozoan T. vaginalis or other microorganisms (Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Candida albicans) used. The present LAMP assay can be performed directly without prior DNA extraction, making the assay an easy, fast, cheap, specific and sensitive diagnostic tool for the detection of T. tenax at the point-of-care of both medical and veterinary clinics in developed and developing countries.

Oral Colonization by Entamoeba gingivalis and Trichomonas tenax: A PCR-Based Study in Health, Gingivitis, and Periodontitis.

Background: The etiology of periodontitis remains unclear, as is the place of gingivitis in its pathophysiology. A few studies linked the colonization by oral parasites (Entamoeba gingivalis and Trichomonas tenax) to periodontal disease and its severity. The aim of the current study was to estimate the prevalence of these oral parasites among healthy individuals, and in patients with gingivitis and periodontitis in Jordan. Methods: The study was conducted during July 2019-December 2019. Samples were composed of saliva and periodontal material including dental plaque sampled with probes. The detection of oral parasites was done using conventional polymerase chain reaction (PCR). Results: The total number of study participants was 237: healthy (n=94), gingivitis (n=53) and periodontitis (n=90). The prevalence of E. gingivalis was 88.9% among the periodontitis patients, 84.9% among the gingivitis patients and 47.9% in the healthy group. For T. tenax, the prevalence was 25.6% among the periodontitis patients, 5.7% among the gingivitis patients and 3.2% in the heathy group. Positivity for E. gingivalis was significantly correlated with the presence of periodontal disease compared to the healthy group with odds ratio (OR) of 6.6. Periodontal disease was also correlated with lower monthly income (OR=8.2), lack of dental care (OR=4.8), and history of diabetes mellitus (OR=4.5). Colonization by E. gingivalis was correlated with gingivitis (OR=6.1) compared to the healthy group. Colonization by E. gingivalis and T. tenax were significantly correlated with periodontitis (OR=6.4 for E. gingivalis, and OR=4.7, for T. tenax) compared to the healthy group. T. tenax was only detected among individuals with generalized periodontal disease compared to its total absence among those with localized disease (19.6% vs. 0.0%; p=0.039). The co-infection rate by the two oral parasites was 11.0%. Conclusions: The higher prevalence of human oral parasites in periodontal disease compared to healthy individuals appears to be more than a mere marker for the disease and might also be associated with disease severity and potential for progression. Thus, the dogmatic view of E. gingivalis and T. tenax as commensals needs to be re-evaluated and their contribution to pathophysiology of periodontal diseases cannot be neglected.