Isolation of Fungi and Gram Negative Bacteria from Toothbrushes and Bathroom Bioaerosols

Rommie Lucia Merino-Alado, Alessandra Garcés, Elisa Chianale, Carlos Corcuera, Wanda El Fakih, Dolly Galviz, Lizbeth Ortiz, Alfredo Sucre Campins, Gustavo Moronta, Elsi Briceño, Maria Eugenia Landaeta, Sofia Mata-Essayag


Objective: To identify, using phenotypic methods, FGNB, NFGNB and Candida sp. in toothbrushes, and environmental samples of bathroom air in a group of students from the Dentistry School of the Universidad Central de Venezuela. Material and Methods: Thirty-four toothbrushes were supplied to the cohort during a 60-day period; environmental samples were collected in the rooms where toothbrushes were kept during this period. All samples were processed by traditional methods of microbiological counting isolation and phenotypic identification using selective and differential agar based on the international guidelines of the United States Pharmacopeia (USP) 38. Results: 82.36% of the toothbrush samples were positive to bacteria and fungi and 91.17% of the environmental samples were positive to enterobacteria. Conclusion: It is necessary to establish antiseptic protocols for the management, storage and disinfection of toothbrushes. The high rate of contamination may represent an opportunity for enterobacteria colonization of oral biofilms, reservoir to infection foci and metastatic infections in certain populations.


Toothbrushing; Gram-Negative Bacteria; Enterobacteriaceae

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Frazelle MR, Munro CL. Toothbrush contamination: A review of the literature. Nurs Res Pract 2012; 2012:420630. doi: 10.1155/2012/420630.

Pesevska S, Ivanovski K, Mindova S, Kaftandzieva A, Ristoska S, Stefanovska E, et al. Bacterial contamination of the toothbrushes. J Int Dent Med Res 2016; 9(1):6-12.

Onourah S, Obika I. Bacterial contamination of used manual toothbrushes obtained from some students of Nnamdi Azikiwe University Awka, Nigeria. Univ J Microbiol Res 2015; 3(4):56-9. doi: 10.13189/ujmr.2015.030404.

Warren DP, Goldschmit MC, Thompson MB, Adler-Storthz K., Keene HJ. The effects of toothpastes on the residual microbial contamination of toothbrushes. J Am Dent Assoc 2001; 132(9):1241-5. doi: 10.14219/jada.archive.2001.0366.

Barker J, Jones MV. The potential spread of infection caused by aerosol contamination of surfaces after flushing a domestic toilet. J Appl Microbiol 2005; 99(2):339-47. doi: 10.1111/j.1365-2672.2005.02610.x.

Nelson-Filho P, Isper AR, Assed S, Faria G, Ito IY. Effect of triclosan dentrifice on toothbrush contamination. Pediatr Dent 2004; 26(1):11-6.

Pathak S, Gopal K. Prevalence of bacterial contamination with antibiotic resistance and enterotoxigenic fecal coliforms in treated drinking water. J Toxicol Environ Health A 2008; 71(7):427-33. doi: 10.1080/15287390701838796.

Paterson D, Bonomo R. Extended-spectrum β-lactamases: A clinical update. Clin Microbiol Rev 2005; 18(4):657-86. doi: 10.1128/CMR.18.4.657-686.2005.

The United States Pharmacopeial Convention. USP 38. Rockville: United States Pharmacopoeial Convention, 2012.

Raiyani CM, Arora R, Bhayya DP, Dogra S, Katageri AA, Singh V. Assessment of microbial contamination on twice a day used toothbrushes head after 1 month and 3 months: An in vitro study. J Nat Sci Biol Med 2015; 6(1):s44-s48. doi: 10.4103/0976-9668.166072.

Eaton KA, Carlile MJ. Tooth brushing behavior in Europe: Opportunities for dental public health. Int Dent J 2008; 58:287-9. doi: 10.1111/j.1875-595X.2008.tb00206.x.

Dayoub MB, Rusilko D, Gross A. Microbial contamination of toothbrushes. J Dent Res 1977; 56(6):706. doi: 10.1177/00220345770560063501.

Marcano C. El cepillo de dientes en la ecología de Candida albicans. Mycopathologia 1981; 74(3):135-41. doi: 10.1007/BF00437155.

Jaimes A, Hernandez F, Martinez E, Rodriguez A, Arenas R. Portadores de Candida sp. en la mucosa oral: tipificación de 35 cepas con CHROMAgar Candida. Med Int Mex 2008; 24(4):262-6.

Naik R, Mujib A, Telagi N, Anil BS, Spoorthi BR. Contaminated tooth brushes-potential threat to oral and general health. J Family Med Prim Care 2015; 4(3):444-8. doi: 10.4103/2249-4863.161350.

Basman A, Peker I, Akca G, Alkurt M, Sarikir C, Celik I. Evaluation of toothbrush disinfection via different methods. Braz Oral Res 2016; 30: doi: 10.1590/1807-3107BO R-2016.vol30.0006.

Ferreira CA, Dagostim Savi G, Panatto AP, da Silva Generoso J, Baricello T. Microbiological evaluation of bristles of frequently used toothbrushes. Dental Press J Orthod 2012; 17(4):72-6. doi: 10.1590/S2176-94512012000400016.

Darlow HM, Bale WR. Infectious hazards of water-closets. Lancet 1959; 1:1196e1200.

Gerba CP, Wallis C, Melnick JL. Microbiological hazards of household toilets: droplet production and the fate of residual organisms. Appl Microbiol 1975; 30:229e237.

Best EL, Sandoe JAT, Wilcox MH. Potential for aerosolization of Clostridium difficile after flushing toilets: the role of toilet lids in reducing environmental contamination risk. J Hosp Infect 2012; 80(1):1-5. doi: 10.1016/j.jhin.2011.08.010.

Proctor CR, Reimann N, Vriens B, Hammes F. Biofilms in shower houses. Water Res 2017; 131:274-86. doi: 10.1016/j.watres.2017.12.027.

Leung WK, Yaw JY, Cheung BP, Jin LJ, Zee KY, et al. Oral colonization by aerobic and facultatively anaerobic gram-negative rods and yeast in Tibetans living in Lhasa. Arch Oral Biol 2003; 48(2):117-23. doi: 10.1016/S0003-9969(02)00201-7.

Sushma R, Karpagaselvi S, Jayalakshmi K, Lokesh P, Jeevan MB. Oral carriage of enterobacteriaceae among school children with chronic nail biting habit. J Oral Maxillofac Pathol 2013; 17(2):163-8. doi: 10.4103/0973-029X.119743.

Xu H, Dongari-Bagtzoglou A. Shaping the oral mycobiota: interactions of opportunistic fungi with oral bacteria and the host. Curr Opin Microbiol 2015; 26:65-70. doi: 10.1016/j.mib.2015.06.002.

Leung WK, Jim L, Joyce YY, Sun Q, Corbert E. Microflora cultivable from minocycline strips placed in persisting periodontal pockets. Arch Oral Biol 2005; 50(1):39-48. doi: 10.1016/j.archoralbio.2004.08.002.

Conti S, dos Santos S, Koga-Ito C, Jorge A. Enterobacteriaceae and Pseudomonadaceae on the dorsum of the human tongue. J Appl Oral Sci 2009; 17(5):375-80. doi: 10.1590/S1678-77572009000500005.

Kuc IM, Samaranayake LP, Van Heyst EN. Oral health and microflora in an institutionalized elderly population in Canada. Int Dent J 1999; 49(1):33-40. doi: 10.1111/j.1875-595X.1999.tb00505.x.

Jobbins J, Baqq J, Parsons K, Finlay I, Addy M, Newcombe RG. Oral carriage of yeast, coliforms and staphylococci in patients with advanced malignant disease. J Oral Pathol Med 1992; 21(7):305-8. doi: 10.1111/j.1600-0714.1992.tb01016.x.

Millership SE, Patel N, Chattopadhyay B. The colonization of patients in an intensive treatment unit with gram-negative flora: the significance of the oral route. J Hosp Infect 1986; 7(3):226-35.

Samaranayake LP, Calman KC, Ferguson MM, Kaye SB, MacFarlane TW, Main B. Wlsh J, Willox J. The oral carriage of yeasts and coliforms in patients on cytotoxic therapy. J Oral Pathol 1984; 13(4):390-3.

Amer A, Galvin S, Healy CM, Moran GP. The microbiome of potentially malignant oral leuokoplakia exhibits enrichment for Fusobacterium, Lelptotrichia, Campylobacter and Rothia species. Front Microbiol 2017; 1(8):2391. doi: 10.3389/fmicb.2017.02391.