Detection of Multidrug resistant cariogenic bacteria among Diabetic Patients attending a Tertiary Hospital in Ado Ekiti, South-Western, Nigeria
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Published:
Oct 17, 2023
Keywords:
Antimicrobial resistance, Diabetics, Streptococcus mitis, Streptococcus mutans, Streptococcus pyogenes, Streptococcus salivarius
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Abstract
Microorganisms colonizing the oral cavity have a significant probability of spreading on epithelial surfaces to neighboring sites. Many reports have associated oral bacteria to oral infectious diseases as well as systemic diseases such as cardiovascular disease and diabetes. Oral care is usually neglected in the control of health problems linked with diabetes which may contribute to the increased morbidity and complications from oral health problems. The study aimed to identify the resident microbial flora and assess the possible carriage of cariogenic bacteria among people with diabetes in Ado- Ekiti, Nigeria. Oral swabs were collected from 306 consenting patients. Culture-based isolation was done using standard techniques on Mitis salivarius agar supplemented with 1% Potassium Tellurite, 10% chocolate agar and MacConkey agar at 37 oC for 18-48 hours. The susceptibility patterns of the isolates were detected using standard techniques as recommended by CLSI. Selected bacteria isolates were genomically identified using the bacterial universal primer 16S rRNA and STR-1 and STR 2 primers (partial tuf genes). Amplified genes were sequenced and phylogenetic analysis was carried out using MEGA 7 software. A total of 447 microbial isolates were identified among which were Staphylococcus spp (14.5%), Klebsiella pneumoniae (3.4%), Escherichia coli (2.5%), Candida spp (8%) while the streptococcal organisms such as Streptococcus pyogenes (35.2%), Streptococcus mitis (14%), Streptococcus mutans (7.3%)
and Streptococcus salivarius (15.1%). Streptococcus mutans was susceptible to meropenem (71.4%) but highly resistant to Penicillin (100%), cefuroxime (71.4%) cefotaxime (68.6%). Streptococcus mitis was highly resistant to Amoxicillin/Clavulanate (100%), Cloxacillin (100%) and Cefuroxime (70.1%) but was found susceptible to Meropenem (77.3%). 16S ribosomal RNA gene amplification and subsequent sequencing of the isolates revealed twenty-six (26) de-novo bacterial strains. In conclusion, the study showed that diabetic individuals carry multidrug-resistant cariogenic bacteria that may predispose them to other oral diseases. There is a need to incorporate routine oral microbiological screening into routine care for diabetic patients in Nigeria.
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Adewumi, F. A., Adegoke, A. A., Ighodalo, U. L., Ojerinde, A. K., Ibeh, I., & Okiki, P. A. (2023). Detection of Multidrug resistant cariogenic bacteria among Diabetic Patients attending a Tertiary Hospital in Ado Ekiti, South-Western, Nigeria. ABUAD International Journal of Natural and Applied Sciences, 3(2), 61-69. https://doi.org/10.53982/aijnas.2023.0302.09-j
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References
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Han, Y. W., & Wang, X. (2013). Mobile microbiome: oral bacteria in extra-oral infections and inflammation. Journal of Dental Research, 92(6), 485–491. https://doi.org/10.1177/0022034513487559
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Khader, Y. S., Albashaireh, Z. S. M., & Hammad, M. M. (2008). Periodontal status of type 2 diabetics compared with nondiabetics in north Jordan. Eastern Mediterranean Health Journal, 14(3), 654–661.
Kilian, M., Chapple, I. L. C., Hannig, M., Marsh, P. D., Meuric, V., Pedersen, A. M. L., Tonetti, M. S., Wade, W. G., & Zaura, E. (2016). The oral microbiome - an update for oral healthcare professionals. British Dental Journal, 221(10), 657–666. https://doi.org/10.1038/SJ.BDJ.2016.865
Lemos, J. A., Palmer, S. R., Zeng, L., Wen, Z. T., Kajfasz, J. K., Freires, I. A., Abranches, J., & Brady, L. J. (2019). The Biology of Streptococcus mutans. Microbiology Spectrum, 7(1). https://doi.org/10.1128/MICROBIOLSPEC.GPP3-0051-2018
Lydia Rajakumari, M., & Saravana Kumari, P. (2016). Prevalence of the bacterial flora in the oral cavity of diabetic individuals in Pondicherry. International Journal of Pharma and Bio Sciences, 7(1), B432–B437.
Marsh D, P., Lewis A O, M., Rogers, H., Williams W, D., & Wilson, M. (2016). Marsh and Martin’s Oral Microbiology. 261. www.elsevierhealth.com
Martineau, F., Picard, F. J., Roy, P. H., Ouellette, M., & Bergeron, M. G. (1998). Species-specific and ubiquitous-DNA-based assays for rapid identification of Staphylococcus aureus. Journal of Clinical Microbiology, 36(3), 618–623. https://doi.org/10.1128/JCM.36.3.618-623.1998
Meng, P., Lu, C., Zhang, Q., Lin, J., & Chen, F. (2017). Exploring the Genomic Diversity and Cariogenic Differences of Streptococcus mutans Strains Through Pan-Genome and Comparative Genome Analysis. Current Microbiology, 74(10), 1200–1209. https://doi.org/10.1007/S00284-017-1305-Z
Murray, E. J., Baron, J. H., Jorgenson, M. A., Pfaller, R. H., & Yolken. (2003). Manual of clinical microbiology 8th edition. Diagnostic Microbiology and Infectious Disease, 47(4), 625. https://doi.org/10.1016/S0732-8893(03)00160-3
Mustard, J. F., & Packham, M. A. (1977). Platelets and diabetes mellitus. The New England Journal of Medicine, 297(24), 1345–1347. https://doi.org/10.1056/NEJM197712152972408
Nazir, M. A. (2017). Prevalence of periodontal disease, its association with systemic diseases and prevention. International Journal of Health Sciences, 11(2), 72. /pmc/articles/PMC5426403/
Ochei, J., & Kolhatkar, A. (2009). Medical Laboratory Science : Theory And Practice. Tata McGraw-Hill Education. https://books.google.com.et/books?id=lciNs3VQPLoC&printsec=copyright#v=onepage&q&f=false
Offenbacher, S., Jared, H. L., O’Reilly, P. G., Wells, S. R., Salvi, G. E., Lawrence, H. P., Socransky, S. S., & Beck, J. D. (1998). Potential pathogenic mechanisms of periodontitis associated pregnancy complications. Annals of Periodontology, 3(1), 233–250. https://doi.org/10.1902/ANNALS.1998.3.1.233
Picard, F. J., Ke, D., Boudreau, D. K., Boissinot, M., Huletsky, A., Richard, D., Ouellette, M., Roy, P. H., & Bergeron, M. G. (2004). Use of tuf Sequences for Genus-Specific PCR Detection and Phylogenetic Analysis of 28 Streptococcal Species. Journal of Clinical Microbiology, 42(8), 3686. https://doi.org/10.1128/JCM.42.8.3686-3695.2004
Saeedi, P., Petersohn, I., Salpea, P., Malanda, B., Karuranga, S., Unwin, N., Colagiuri, S., Guariguata, L., Motala, A. A., Ogurtsova, K., Shaw, J. E., Bright, D., & Williams, R. (2019). Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas, 9th edition. Diabetes Research and Clinical Practice, 157, 107843. https://doi.org/10.1016/j.diabres.2019.107843
Santiago, K. B., Piana, G. M., Conti, B. J., Cardoso, E. de O., Andrade, B. F. M. T., Zanutto, M. R., Rall, V. L. M., Fernandes, A., & Sforcin, J. M. (2018). Microbiological control and antibacterial action of a propoliscontaining mouthwash and control of dental plaque in humans. Natural Product Research, 32(12), 1441–1445. https://doi.org/10.1080/14786419.2017.1344664
Sarkonen, N., Könönen, E., Eerola, E., Könönen, M., Jousimies-Somer, H., & Laine, P. (2005). Characterization of Actinomyces species isolated from failed dental implant fixtures. Anaerobe, 11(4), 231–237. https://doi.org/10.1016/J.ANAEROBE.2005.01.002
Seymour, GJ, PJ, F., MP, C., S, L., & K, Y. (2007). Relationship between periodontal infections and systemic disease. Clinical Microbiology and Infection : The Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, 13 Suppl 4(SUPPL. 2), 2–6. https://doi.org/10.1111/J.1469-0691.2007.01798.X
Taylor, G. W. (2001). Bidirectional interrelationships between diabetes and periodontal diseases: an epidemiologic perspective. Annals of Periodontology, 6(1), 99–112. https://doi.org/10.1902/ANNALS.2001.6.1.99
Ussar, S., Fujisaka, S., & Kahn, C. R. (2016). Interactions between host genetics and gut microbiome in diabetes and metabolic syndrome. Molecular Metabolism, 5(9), 795. https://doi.org/10.1016/J.MOLMET.2016.07.004
WHO (2022). Oral health. https://www.who.int/news-room/fact-sheets/detail/oral-health
Wild, S., Roglic, G., Green, A., Sicree, R., & King, H. (2004). Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care, 27(5), 1047–1053. https://doi.org/10.2337/DIACARE.27.5.1047
Willey, J. M., Sherwood, L. M., & Christopher J. Woolverton. (2013). Microbiology, Prescott.
Zarco, M. F., Vess, T. J., & Ginsburg, G. S. (2012). The oral microbiome in health and disease and the potential impact on personalized dental medicine. Oral Diseases, 18(2), 109–120. https://doi.org/10.1111/J.1601-0825.2011.01851.
American Diabetes Association. (2009). Diagnosis and Classification of Diabetes Mellitus. Diabetes Care, 32(Supplement_1), S62–S67. https://doi.org/10.2337/DC09-S062
Awano, S., Ansai, T., Takata, Y., Soh, I., Akifusa, S., Hamasaki, T., Yoshida, A., Sonoki, K., Fujisawa, K., & Takehara, T. (2008). Oral health and mortality risk from pneumonia in the elderly. Journal of Dental Research, 87(4), 334–339. https://doi.org/10.1177/154405910808700418
Beck, J. D., & Offenbacher, S. (2005). Systemic Effects of Periodontitis: Epidemiology of Periodontal Disease and Cardiovascular Disease. Journal of Periodontology, 76 Suppl 11S(11 Suppl), 2089–2100. https://doi.org/10.1902/jop.2005.76.11-S.2089
Chapple, I. L. C., & Genco, R. (2013). Diabetes and periodontal diseases: consensus report of the Joint EFP/AAP Workshop on Periodontitis and Systemic Diseases. Journal of Periodontology, 84(4 Suppl), S106–S112. https://doi.org/10.1902/JOP.2013.1340011
Clinical Laboratory Standard Institue (CLSI). (2018). Performance Standards for Antimicrobial Susceptibility Testing. 26th ed. CLSI supplement M100S. In Clinical and Laboratory Standards Institute, Wayne, PA.
Cui, L., Morris, A., & Ghedin, E. (2013). The human mycobiome in health and disease. Genome Medicine, 5(7), 1–12. https://doi.org/10.1186/GM467/FIGURES/4
Davidovich, N. V., Galieva, A. S., Davydova, N. G., Malygina, O. G., Kukalevskaya, N. N., Simonova, G. V., & Bazhukova, T. A. (2020). Spectrum and resistance determinants of oral streptococci clinical isolates. Klinicheskaia Laboratornaia Diagnostika, 65(10), 632–637. https://doi.org/10.18821/0869-2084-2020-65-10-632-637
Dewhirst, F. E., Chen, T., Izard, J., Paster, B. J., Tanner, A. C. R., Yu, W. H., Lakshmanan, A., & Wade, W. G. (2010). The human oral microbiome. Journal of Bacteriology, 192(19), 5002–5017. https://doi.org/10.1128/JB.00542-10
Douglass, C. W. (2006). Risk assessment and management of periodontal disease. Journal of the American Dental Association (1939), 137 Suppl(11 SUPPL.), S27–S32. https://doi.org/10.14219/JADA.ARCHIVE.2006.0410
Gao, L., Xu, T., Huang, G., Jiang, S., Gu, Y., & Chen, F. (2018). Oral microbiomes: more and more importance in oral cavity and whole body. Protein & Cell, 9(5), 488–500. https://doi.org/10.1007/S13238-018-0548-1
Genco, R. J., Grossi, S. G., Ho, A., Nishimura, F., & Murayama, Y. (2005). A Proposed Model Linking Inflammation to Obesity, Diabetes, and Periodontal Infections. Journal of Periodontology, 76 Suppl 11S(11 Suppl), 2075–2084. https://doi.org/10.1902/jop.2005.76.11-S.2075
Han, Y. W., & Wang, X. (2013). Mobile microbiome: oral bacteria in extra-oral infections and inflammation. Journal of Dental Research, 92(6), 485–491. https://doi.org/10.1177/0022034513487559
Joshipura, K. J., Rimm, E. B., Douglass, C. W., Trichopoulos, D., Ascherio, A., & Willett, W. C. (1996). Poor oral health and coronary heart disease. Journal of Dental Research, 75(9), 1631–1636. https://doi.org/10.1177/00220345960750090301
Ke, D., Picard, F. J., Martineau, F., Ménard, C., Roy, P. H., Ouellette, M., & Bergeron, M. G. (1999). Development of a PCR assay for rapid detection of enterococci. Journal of Clinical Microbiology, 37(11), 3497–3503. https://doi.org/10.1128/JCM.37.11.3497-3503.1999
Khader, Y. S., Albashaireh, Z. S. M., & Hammad, M. M. (2008). Periodontal status of type 2 diabetics compared with nondiabetics in north Jordan. Eastern Mediterranean Health Journal, 14(3), 654–661.
Kilian, M., Chapple, I. L. C., Hannig, M., Marsh, P. D., Meuric, V., Pedersen, A. M. L., Tonetti, M. S., Wade, W. G., & Zaura, E. (2016). The oral microbiome - an update for oral healthcare professionals. British Dental Journal, 221(10), 657–666. https://doi.org/10.1038/SJ.BDJ.2016.865
Lemos, J. A., Palmer, S. R., Zeng, L., Wen, Z. T., Kajfasz, J. K., Freires, I. A., Abranches, J., & Brady, L. J. (2019). The Biology of Streptococcus mutans. Microbiology Spectrum, 7(1). https://doi.org/10.1128/MICROBIOLSPEC.GPP3-0051-2018
Lydia Rajakumari, M., & Saravana Kumari, P. (2016). Prevalence of the bacterial flora in the oral cavity of diabetic individuals in Pondicherry. International Journal of Pharma and Bio Sciences, 7(1), B432–B437.
Marsh D, P., Lewis A O, M., Rogers, H., Williams W, D., & Wilson, M. (2016). Marsh and Martin’s Oral Microbiology. 261. www.elsevierhealth.com
Martineau, F., Picard, F. J., Roy, P. H., Ouellette, M., & Bergeron, M. G. (1998). Species-specific and ubiquitous-DNA-based assays for rapid identification of Staphylococcus aureus. Journal of Clinical Microbiology, 36(3), 618–623. https://doi.org/10.1128/JCM.36.3.618-623.1998
Meng, P., Lu, C., Zhang, Q., Lin, J., & Chen, F. (2017). Exploring the Genomic Diversity and Cariogenic Differences of Streptococcus mutans Strains Through Pan-Genome and Comparative Genome Analysis. Current Microbiology, 74(10), 1200–1209. https://doi.org/10.1007/S00284-017-1305-Z
Murray, E. J., Baron, J. H., Jorgenson, M. A., Pfaller, R. H., & Yolken. (2003). Manual of clinical microbiology 8th edition. Diagnostic Microbiology and Infectious Disease, 47(4), 625. https://doi.org/10.1016/S0732-8893(03)00160-3
Mustard, J. F., & Packham, M. A. (1977). Platelets and diabetes mellitus. The New England Journal of Medicine, 297(24), 1345–1347. https://doi.org/10.1056/NEJM197712152972408
Nazir, M. A. (2017). Prevalence of periodontal disease, its association with systemic diseases and prevention. International Journal of Health Sciences, 11(2), 72. /pmc/articles/PMC5426403/
Ochei, J., & Kolhatkar, A. (2009). Medical Laboratory Science : Theory And Practice. Tata McGraw-Hill Education. https://books.google.com.et/books?id=lciNs3VQPLoC&printsec=copyright#v=onepage&q&f=false
Offenbacher, S., Jared, H. L., O’Reilly, P. G., Wells, S. R., Salvi, G. E., Lawrence, H. P., Socransky, S. S., & Beck, J. D. (1998). Potential pathogenic mechanisms of periodontitis associated pregnancy complications. Annals of Periodontology, 3(1), 233–250. https://doi.org/10.1902/ANNALS.1998.3.1.233
Picard, F. J., Ke, D., Boudreau, D. K., Boissinot, M., Huletsky, A., Richard, D., Ouellette, M., Roy, P. H., & Bergeron, M. G. (2004). Use of tuf Sequences for Genus-Specific PCR Detection and Phylogenetic Analysis of 28 Streptococcal Species. Journal of Clinical Microbiology, 42(8), 3686. https://doi.org/10.1128/JCM.42.8.3686-3695.2004
Saeedi, P., Petersohn, I., Salpea, P., Malanda, B., Karuranga, S., Unwin, N., Colagiuri, S., Guariguata, L., Motala, A. A., Ogurtsova, K., Shaw, J. E., Bright, D., & Williams, R. (2019). Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas, 9th edition. Diabetes Research and Clinical Practice, 157, 107843. https://doi.org/10.1016/j.diabres.2019.107843
Santiago, K. B., Piana, G. M., Conti, B. J., Cardoso, E. de O., Andrade, B. F. M. T., Zanutto, M. R., Rall, V. L. M., Fernandes, A., & Sforcin, J. M. (2018). Microbiological control and antibacterial action of a propoliscontaining mouthwash and control of dental plaque in humans. Natural Product Research, 32(12), 1441–1445. https://doi.org/10.1080/14786419.2017.1344664
Sarkonen, N., Könönen, E., Eerola, E., Könönen, M., Jousimies-Somer, H., & Laine, P. (2005). Characterization of Actinomyces species isolated from failed dental implant fixtures. Anaerobe, 11(4), 231–237. https://doi.org/10.1016/J.ANAEROBE.2005.01.002
Seymour, GJ, PJ, F., MP, C., S, L., & K, Y. (2007). Relationship between periodontal infections and systemic disease. Clinical Microbiology and Infection : The Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, 13 Suppl 4(SUPPL. 2), 2–6. https://doi.org/10.1111/J.1469-0691.2007.01798.X
Taylor, G. W. (2001). Bidirectional interrelationships between diabetes and periodontal diseases: an epidemiologic perspective. Annals of Periodontology, 6(1), 99–112. https://doi.org/10.1902/ANNALS.2001.6.1.99
Ussar, S., Fujisaka, S., & Kahn, C. R. (2016). Interactions between host genetics and gut microbiome in diabetes and metabolic syndrome. Molecular Metabolism, 5(9), 795. https://doi.org/10.1016/J.MOLMET.2016.07.004
WHO (2022). Oral health. https://www.who.int/news-room/fact-sheets/detail/oral-health
Wild, S., Roglic, G., Green, A., Sicree, R., & King, H. (2004). Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care, 27(5), 1047–1053. https://doi.org/10.2337/DIACARE.27.5.1047
Willey, J. M., Sherwood, L. M., & Christopher J. Woolverton. (2013). Microbiology, Prescott.
Zarco, M. F., Vess, T. J., & Ginsburg, G. S. (2012). The oral microbiome in health and disease and the potential impact on personalized dental medicine. Oral Diseases, 18(2), 109–120. https://doi.org/10.1111/J.1601-0825.2011.01851.