Khalid et al - Al Manhal

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protozoa (Entamoeba gingivalis and Trichomonas tenax) which can cause or initiate oral ... gingivalis and/or Trichomonas tenax) were detected in 11 individuals ...
Khalid et al - Commensal Oral Protozoa among Patients Attending Academic Dental Teaching Hospital......

Original Article Commensal Oral Protozoa among Patients Attending Academic Dental Teaching Hospital, Khartoum State, Sudan Arwa Khalid1, El-Amin Abdulkareem1, Eman Kheir1, Alfatih Aljafari2* 1

Department of Medical Parasitology, Faculty of Medical Laboratory Sciences, Al-Neelain University; Khartoum-Sudan, and 2Department of Pathology, College of Medicine, Aljouf University; Sakaka, Kingdom of Saudi Arabia. *Corresponding author: [email protected] Abstract Background: Poor oral hygiene provides a suitable condition for presence and multiplication of commensal oral protozoa (Entamoeba gingivalis and Trichomonas tenax) which can cause or initiate oral problems. The aim: This study aimed at determining the frequency of oral commensal protozoa and their relationship with oral hygiene and dental problems. Patients and Methods: This is a case-control study involving 29 patients attending Academic Dental Teaching Hospital with oral problems and 31 apparently healthy normal individuals as control group. Saliva and gingival swabs were collected from each participant. Samples were examined microscopically for presence of motile protozoa and cultured in Locke-Egg (LE) media. Results: Oral protozoa (Entamoeba gingivalis and/or Trichomonas tenax) were detected in 11 individuals (37.9%) of the cases group. Five samples were positive by direct wet preparation and 11 positive samples detected by culture. The sensitivity and specificity for wet preparation were 17% and 100% and for the LE culture were 38% and 100%. No protozoon was detected among the healthy control group. The presence of oral protozoa correlated a bad oral hygiene, presence of gingivitis and periodontitis. Conclusions: Gingivitis and periodontitis among the studied Sudanese patients could have furnished suitable conditions for the presence and growth of these protozoa and/or the protozoal presence could have caused/aggravated gingivitis and periodontitis. Confirmation of a causal relationship necessitates further studies. Khalid A, Abdulkareem E, Kheir E, Aljafari A. Commensal Oral Protozoa among Patients Attending Academic Dental Teaching Hospital, Khartoum State, Sudan. AUMJ, 2015 June 1; 2(2): 17 - 20. Keywords: Entamoeba gingivalis, Trichomonas tenax, oral hygiene, Sudan. Introduction Entamoeba gingivalis (E. gingivalis) and Trichomonas tenax (T. tenax) are the protozoa which normally live in the oral cavity(1). Their presence in oral cavity is considered as a sign of poor oral hygiene. E. gingivalis is morphologically very similar to E. histolytica. It lives on the surface of teeth, gums and some times in the tonsilar crypts. They become often abundant in case of gum or tonsils disease. But until now, no evidence shows they cause these conditions. E. gingivalis feeds on bacteria, food particles, dead epithelial cells and leukocytes. The ingested leukocytes facilitate the identification of E. gingivalis because this is the only species that ingests these cells(2). Due to the high incidence of E. gingivalis is associated to periodontal diseases, some authors suggests that this amoeba might have a pathogenetic role in this condition(3), or could be opportunistic(4). T. tenax has only trophozoite stage. The later is an elongated cell 5 - 16 µm long by 2 - 15 µm wide(5). Transmission can be through saliva, droplet spray,

kissing and use of contaminated dishes, spoons, food and drinking water. T. tenax live around the teeth most abundant between the teeth and gum, tooth cavities, crypts of tonsils and in submaxillary glands(6). apparently, cannot survive passage through the digestive tract. It is quite resistant to change in temperature and survives for several hours in drinking water(7). Since the organism is believed to enter the respiratory tract by aspiration from the oropharynx and cause bronchopulmonary trichomoniasis, the importance of oral infections has been highlighted (2, 8, 9) . Since no previous such studies, we planned to determine the extent of the presence of these protozoa in oral cavity of Sudanese dental patients by wet mount and LE culturing and to correlate their presence with oral hygiene or systemic diseases. Patients and Methods This is a case-control study in which 29 patients (10 males and 19 female; age range (24 - 67 years) attended Khartoum Dental Hospital was enrolled

Aljouf University Medical Journal (AUMJ), 2015 June 1; 2(2): 17 -20.

2015

Khalid et al - Commensal Oral Protozoa among Patients Attending Academic Dental Teaching Hospital......

together with 31 apparently healthy individuals as control (17 male and 14 female; age range 23 - 58 years). Patients with diabetes, chronic liver and kidney diseases and HIV/AIDS, on immunosuppressant medications and severely anemic were excluded from this study. Saliva and gingival swab samples were collected from participants following relevant quality control and safety measures of sample collection. Saliva samples were examined by direct wet preparation to detect the motile trophozoite. The local Khartoum Dental Hospital ethical committee approved the research and an informed consent was collected each participant. Culture media and conditions: Preparation of LockeEgg (LE) culture medium: 500 mg of starch powder was dried at 150 °C for 2.5 hrs, cooled and suspended in 9.5 mL sterile distilled water by vortexing. 1 mL aliquots are diluted with 9 mL of sterile distilled water and kept refrigerated. Before use, resuspended starch by vortexing and pipette the desired volume into culture tubes with medium. 0.2 ml (1 mg) is often a suitable amount to add. Preparation of egg slant: Add 12.5 mL Lock´s solution (re-autoclaved sterile solution containing 8 g sodium chloride, 0.2 g calcium chloride, 0.2 g potassium chloride, 0.01 g magnesium chloride 2 g disodium hydrogen phosphate, 0.4 g sodium bicarbonate, 0.3 g potassium dihydrogen phosphate per liter) per 45 mL of egg (70% ethanol surface sterilized fresh hens' eggs were broken into a graduate cylinder). The mixture was emulsified and filtered though sterile gauze into a sterile flask. 5 mL aliquots were dispensed into sterile culture tubes and incubated in water path in 100 ºC for 15 minutes with the tubes at an angle to produce a slope free of bubbles. After cooling to room temperature, 6 mL of Lock´s solution and 0.2 mL of the sterile diluted starch solution were added to each of the egg slant tubes. Tubes were autoclave (15 minutes, 121ºc, 15 1bs), cooled to room temperature and were caps were tighten and store refrigerated for up to 6 months(10). All swabs and saliva samples were inoculated in LE medium and incubated at ~35 oC for 4 days and culture samples were examined microscopically for presence of motile parasites. Statistical Package for Social Sciences (SPSS version 20) was used for data presentation and Chi-Square test for categorical data comparison analysis with p