The Prevalence of Canine Oral Protozoa and Their Association with Periodontal Disease.

Periodontal disease is one of the most important health concerns for companion animals. Research into canine forms of periodontitis has focused on the identification and characterization of the bacterial communities present. However, other microorganisms are known to inhabit the oral cavity and could also influence the disease process. A novel, broad spectrum 18S PCR was developed and used, in conjunction with next-generation sequencing analyses to target the identification of protists. Trichomonas sp. and Entamoeba sp. were identified from 92 samples of canine plaque. The overall prevalence of trichomonads was 56.52% (52/92) and entamoebae was 4.34% (4/92). Next-generation sequencing of pooled healthy, gingivitis, early-stage periodontitis, and severe periodontitis samples revealed the proportion of trichomonad sequences to be 3.51% (health), 2.84% (gingivitis), 6.07% (early periodontitis), and 35.04% (severe periodontitis), respectively, and entamoebae to be 0.01% (health), 0.01% (gingivitis), 0.80% (early-stage periodontitis), and 7.91% (severe periodontitis) respectively. Both genera of protists were statistically associated with plaque from dogs with periodontal disease. These findings provide the first conclusive evidence for the presence of oral protozoa in dog plaque and suggest a possible role for protozoa in the periodontal disease process.

Growth kinetics, antigen profiling, and proteinase activity of Egyptian Trichomonas tenax isolates derived from patients having oral infections.

The role of Trichomonas tenax as a pathogen had been clearly implicated in various pathological processes that arise outside the boundaries of the mouth. Although a relationship between the increased occurrence of this protozoan and progression of periodontal disease has been demonstrated, the ability of T. tenax in causing oral infections and the precise mechanism of tissue damage is not well known. The present study aimed to investigate different isolates of T.tenax from individuals having oral infections. Plaques and/or calculi samples were collected from 70 individuals who were diagnosed as having periodontitis and/or gingivitis, then subjected to parasitological examination and culture on modified trypticase, yeast and iron medium (TYI-S-33). Isolates successfully maintained in culture were further subjected to analysis of protein profile of lysates by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and analysis of proteinases by non-denaturing gelatin-SDS-PAGE. Comparison of growth kinetics of seven T. tenax isolates showed a wide variability in the growth characteristics. Protein profiles of the seven isolates revealed a total 53 bands ranged in molecular weight (MW) from 5 to 95kDa using 12% resolution gel. Also, T. tenax isolates were found to possess 19 proteinase bands ranged in MW from 14 to 66kDa. The proteolytic bands were intensified by a cysteine proteinase activator and totally disappeared by treatment with a cysteine proteinase inhibitor suggesting that the proteinases were of cysteine proteinases type. The high frequency of T. tenax detected (28.6%) along with the variability in protein profiling and proteolytic activity of the isolates supports the possible pathogenicity of T. tenax and clarifies a conclusion that different strains with possibility of variable pathogenic potential may exist.

Prevalence of oral Entamoeba gingivalis and Trichomonas tenax in patients with periodontal disease and healthy population in Shiraz, southern Iran.

BACKGROUND: It was shown that two parasites of Entamoeba gingivalis (E. gingivalis) and Trichomonas tenax (T. tenax) may be responsible for oral parasitic infection. This study was designed to evaluate the prevalence of these parasites in oral cavity of patients with periodontal disease and in healthy population in Shiraz, Southern Iran. MATERIALS AND METHODS: A total of 50 patients with periodontal disease (case group) and 50 subjects with healthy gingiva (control group) entered in the present study. A questionnaire recorded general health, smoking habits, and any history of antibiotic consumption during the last six months for each patient. In the case group, saliva was collected by sterile swab and the gingival crevicular fluid by the paper point. The plaque and calculi were collected by sterile curette and scaler. In the control group, saliva and gingival crevicular fluid were collected and sent to laboratory for further studies. RESULTS: In the case group, nine patients were infected, six with E. gingivalis and three with T. tenax. Seven patients had mobility of the teeth, one patient was smoker and five had previous history of antibiotic consumption. In the control group, only one subject was infected with E. gingivalis without any history of smoking and antibiotic consumption. CONCLUSION: Parasitic infections are relatively common in patients with periodontal disease. It seems that follow-up of instructions are essential in control of parasitic infection in Southern Iran.

Entamoeba gingivalis y Trichomonas tenax en pacientes diabéticos / Entamoeba gingivalis and Trichomonas tenax in diabetic patients

Objetivo: estudiar la frecuencia de Entamoeba gingivalis y Trichomonas tenax en pacientes diabéticos y la relación, de estos protozoarios, con IgA, pH y las periodontopatías. Material y método: se trabajó con 50 pacientes de ambos sexos, cuyas edades oscilaron entre 25 a 69 años, que concurrieron al servicio de Diagnóstico de la Facultad de Odontología de Rosario, Argentina. De cada paciente se obtuvo una muestra de placa y/o cálculo dental y una de saliva para la búsqueda de E. gingivalis y T. tenax por microscopía directa, cultivo y tinción tricrómica para E. gingivalis. En saliva se determinó también concentración de IgA y pH. Los resultados se analizaron estadísticamente. Resultados: de los 50 pacientes estudiados, 37 (74 por ciento) presentaron parásitos. La prevalencia de E. gingivalis fue 91 por ciento y 32 por ciento para T. tenax. Los cultivos de T. tenax aumentaron significativamente la sensibilidad diagnótica. Sin embargo para E. gingivalis, ni el cultivo ni la coloración aumentaron la sensibilidad. Las determinaciones de IgA salival y pH, en la población de pacientes parasitados, mostraron que un elevado número de los mismos presentaron rangos de valores normales. La patología bucal predominante en los individuos que presentaron parásitos fue gingivitis. Conclusión: la presencia de parásitos no está asociada con ninguna de las tres variables. (AU)

Incidência do Helicobacter Pylori na placa dentária e mucosa gástrica / Incidence of Helicobacter Pylory in dental plaque and gastric mucous

Neste trabalho, foi estudado, através do teste da urease, a incidência do Helicobacter pylori na placa dentária e mucosa gástrica de 80 pacientes adultos submetidos a esôfagogastroduodenoscopia que, por indicação médica, procuraram o serviço de endodscopia digestiva do Instituto Hospitalar General Edson Ramalho em João Pessoa Paraíba-Brasil. Nos resultados obtidos, verificou-se a elevada incidência desta bactéria na placa dentária, com 64 casos positivos, assim como na mucosa gástrica, com 65 casos de positividade. No confronto de presença simultânea, identificou-se a bactéria em 56 casos, dos quais 48 pacientes apresentaram um ou mais tipos de afecções gastroduodenal. Nos pacientes com resultado da endoscopia normal, registrou-se, simultaneamente, 8 casos positivos entre 20 pacientes. Conclui-se assim, que estes locais funcionam como prováveis reservatórios destes microorganismos

Entamoeba gingivalis y trichomonas tenax: hallazgo en poblaciones humanas con y sin patología bucal / Entamoeba gingivalis and trichomonas tenax: finding in human populations with and without buccal pathology

Se estudiaron dos poblaciones de 50 pacientes adultos, de ambos sexos, de la ciudad de Rosario: la población 1 (control) con bocas sanas y buenos hábitos de higiene oral y la población 2 con gingivitis y/o periodontitis y mala higiene bucal. El objetivo fue la búsqueda de Entamoeba gingivalis y Trichomonas tenax para determinar sus prevalencias en individuos con patología oral y la relación de estos parásitos con la IgA y pH salival. De cada paciente se obtuvo una muestra de placa y sarro dental y una de saliva, para examen parasitológico por microscopia directa, 100x400 x. Se colorearon frotis de sarro y placa dental con tinción tricrómica para identificación de E. gingivalis. Se determinó además pH e IgA salival. El 50 por ciento de los pacientes con buena salud bucal presentaron parásitos: 20 monoparasitados con E. gingivalis, 1 con T. tenax y los 4 restantes con ambos protozoos. La población con patología bucal presentó un incremento significativo del porcentaje de parasitosis (78 por ciento) con respecto a la anterior, se presentaron 29 individuos monoparasitados con E. gingivalis, 1 con T. tenax y 9 con ambos protozoos. La frecuencia de E. gingivalis y T. tenax en la población 1 fueron 48 por cienbto y 10 por ciento respectivamente y resultaron notablemente mayores en la población 2 (E. gingivalis 76 por ciento y T. tenax 20 por ciento). Los hallazgos de protozoos en las dos poblaciones predominaron en placa y sarro dental. La utilización de la técnica de tinción tricrómica de Gomori aumentó la posibilidad de diagnóstico en ambas poblaciones. La presencia de parásitos orales fue independiente del valor de IgA en las dos poblaciones. La población control presentó dependencia entre el pH salival y la presencia de protozoos y se podría inferir que el pH ligeramente ácido favorece la colonización parasitaria. En la población con patología bucal no habría dependencia entre estas variables. La alta frecuencia encontrada de estos protozoos en pacientes con gingivitis y/o periodontitis destaca la importancia del diagnóstico para aplicar el tratamiento específico que permite la erradicación parasitaria

Entamoeba gingivalis y trichomonas tenax: hallazgo en poblaciones humanas con y sin patología bucal / Entamoeba gingivalis and trichomonas tenax: finding in human populations with and without buccal pathology

Se estudiaron dos poblaciones de 50 pacientes adultos, de ambos sexos, de la ciudad de Rosario: la población 1 (control) con bocas sanas y buenos hábitos de higiene oral y la población 2 con gingivitis y/o periodontitis y mala higiene bucal. El objetivo fue la búsqueda de Entamoeba gingivalis y Trichomonas tenax para determinar sus prevalencias en individuos con patología oral y la relación de estos parásitos con la IgA y pH salival. De cada paciente se obtuvo una muestra de placa y sarro dental y una de saliva, para examen parasitológico por microscopia directa, 100x400 x. Se colorearon frotis de sarro y placa dental con tinción tricrómica para identificación de E. gingivalis. Se determinó además pH e IgA salival. El 50 por ciento de los pacientes con buena salud bucal presentaron parásitos: 20 monoparasitados con E. gingivalis, 1 con T. tenax y los 4 restantes con ambos protozoos. La población con patología bucal presentó un incremento significativo del porcentaje de parasitosis (78 por ciento) con respecto a la anterior, se presentaron 29 individuos monoparasitados con E. gingivalis, 1 con T. tenax y 9 con ambos protozoos. La frecuencia de E. gingivalis y T. tenax en la población 1 fueron 48 por cienbto y 10 por ciento respectivamente y resultaron notablemente mayores en la población 2 (E. gingivalis 76 por ciento y T. tenax 20 por ciento). Los hallazgos de protozoos en las dos poblaciones predominaron en placa y sarro dental. La utilización de la técnica de tinción tricrómica de Gomori aumentó la posibilidad de diagnóstico en ambas poblaciones. La presencia de parásitos orales fue independiente del valor de IgA en las dos poblaciones. La población control presentó dependencia entre el pH salival y la presencia de protozoos y se podría inferir que el pH ligeramente ácido favorece la colonización parasitaria. En la población con patología bucal no habría dependencia entre estas variables. La alta frecuencia encontrada de estos protozoos en pacientes con gingivitis y/o periodontitis destaca la importancia del diagnóstico para aplicar el tratamiento específico que permite la erradicación parasitaria (AU)

Entamoeba gingivalis in human immunodeficiency virus type 1-infected patients with periodontal disease.

Necrotic periodontal disease is a progressive painful oral lesion in human immunodeficiency virus type 1 (HIV-1)-infected patients, and the etiology is unknown. Earlier studies of HIV-1-infected patients have shown significant changes in the viral and fungal oral microflora. The aim of this study was to relate the occurrence of protozoa to clinical symptoms and immunosuppression. Oral symptoms were registered in 45 patients at different stages of the HIV-1 infection and in 15 HIV-seronegative healthy controls. Saliva and dental plaque were analyzed for the presence of protozoa. Entamoeba gingivalis was the only protozoa found in the oral cavities of HIV-1-infected patients with periodontal disease. Its presence was not related to the degree of immunodeficiency but to the HIV diagnosis. This study describes for the first time the findings of E. gingivalis in the oral cavities of HIV-1 infected patients.

Trichomonas tenax proteolytic activity.

In this study, proteolytic activity of Trichomonas tenax collected directly from patient's dentobacterial plaque was examined. Electrophoretic method involving polyacrylamide gels (Commassie Brilliant Blue R-250) and electrophoretic method involving gelatin-containing polyacrylamide gels, have been used to analyse Trichomonas tenax proteolytic activity. The most obvious and the fastest activities were obtained when gels were incubated in pH 4.6; followed by results of incubating in pH 5.6; while in pH 2.8 activity was less effective but still obvious. Proteolytic activities were the most effective in area of protein MW 36 kDa. Different activities of enzymes depending on pH of incubated media indicate the presence of different endopeptidases in cell lysates of protozoon Trichomonas tenax from dentobacterial plaque.

Specific and sensitive detection of Trichomonas tenax by the polymerase chain reaction.

A polymerase chain reaction (PCR) protocol was developed for specific detection of Trichomonas tenax by using a pair of primers designed for its 18S rRNA gene. The detection was specific for T. tenax, since no amplification was detected with DNAs from Trichomonas vaginalis, which belongs to the same genus as T. tenax, in addition to various species of oral protists, fungi and bacteria, and human leukocytes. This method had a detection limit of 100 fg for T. tenax genomic DNA and could detect T. tenax cells in dental plaque at a concentration of as low as 5 cells per PCR mixture. Direct detection from clinical dental plaque samples was also possible; therefore, the present PCR procedure could provide a simple and rapid detection method of T. tenax in dental plaque.

Detection and identification of Entamoeba gingivalis by specific amplification of rRNA gene.

A pair of oligonucleotide primers were designed from the nucleotide sequence of the gene encoding the small subunit ribosomal RNA (SrRNA) of the oral protozoan parasite Entamoeba gingivalis. The primers amplified a 1.4-kb DNA fragment by polymerase chain reaction and were specific for Entamoeba gingivalis but not for other protozoa, oral protists and bacteria, or human leukocytes. With this method, the DNA from as few as 30 cells of Entamoeba gingivalis could be detected. These results suggest that this approach is applicable to the detection and identification of Entamoeba gingivalis in the human oral cavity.