Sys Rev Pharm 2020; 11(4): 396 400 A multifaceted review journal in the field of pharmacy E-ISSN 0976-2779 P-ISSN 0975-8453 Granulicatella Adiacens Bacteria Isolation from Perodontitical Patients with Polymerase Chain Reaction Techniques Harun Achmad1, Sri Oktawati2,Andi Mardiana Adam2,Burhanuddin Pasiga3, Rizalinda Sjahril4, Aulia Azizah5, Bayu Indra Sukmana6, Huldani7, Heri Siswanto8 , Ingrid Neormansyah8 1Lecturer of Pedodontics Department, Dentistry Faculty, Hasanuddin University, Makassar, Indonesia 2Lecturer of Periodontology Department, Dentistry Faculty, Hasanuddin University, Makassar, Indonesia 3Lecturer of Dental Public Health Department, Dentistry Faculty, Hasanuddin University, Makassar, Indonesia 4 Lecturer of Microbiology Department, Medical Faculty, Hasanuddin University, Makassar, Indonesia 5Department of Preventive and Public Health Dentistry, Faculty of Dentistry, Lambung Mangkurat University, Banjarmasin, Indonesia. 6Department of Dental Radiology, Faculty of Dentistry, Lambung Mangkurat University, Banjarmasin, Indonesia. 7Department of Physiology, Faculty of Medicine, Lambung Mangkurat University, Banjarmasin, Indonesia. 8 Resident of Periodontology Department, Dentistry Faculty, Hasanuddin University, Makassar, Indonesia Correspondence Author E-mail: [email protected] Article History: Submitted: 10.01.2020 Revised: 12.03.2020 Accepted: 24.04.2020 ABSTRACT Background: Periodontitis is an inflammatory disease of dental support the NCBI Gene Bank using BLAST analysis. tissue caused by certain groups of microorganisms, resulting in Result: In this study, Granulicatella adiancense was found in 13 progressive destruction of the periodontal ligament and alveolar bone, samples from 18 solated samples from patients with chronic with pocket formation, recession, or both1. Destruction of periodontal periodontitis and in periodontal pocket depth more than 7 mm. caused by the host defence. Conclusion: Granulicatella adiancense is a normal flora bacteria in the Objective: Granulicatella adiacens is a small Gram-positive, non-motile, oral cavity and chronic periodontitis conditions, the presence of this non-spore-forming, facultatively anaerobic coccus. As a member of bacteria is detected in the deeper periodontal pocket depth. Nutritionally Variant Streptococci, genus Granulicatella, it is a part of Keywords: Chronic periodontitis, DNA squence, Granulicatella the normal microbiota in the oral cavity, urogenital, and intestinal tract. adiacens It may also participate in periodontitis, caries, and endodontic Correspondence: infections. Our aim in this study was to investigate the presence Harun Achmad of Granulicatella adiacens in periodontal disease Lecturer of Pedodontics Department, Dentistry Faculty, Hasanuddin Methods: A total 18 samples from patients with chronic periodontitis University was collected, by using the conventional PCR technique for checking Makassar, Indonesia the presence of bacterial bands, followed by using DNA sequence E-mail: [email protected] methods, to complete the examination of bacteria with Conventional DOI: 10.31838/srp.2020.4.59 PCR which results are used as a basis for nucleotide examination on @Advanced Scientific Research. All rights reserved INTRODUCTION colonies of other bacteria. Isolates have been recovered from Periodontitis is an inflammatory disease of dental support blood, abscesses, oral ulcers, and urethral samples. Because tissue caused by certain groups of microorganisms, resulting of both difficulties in culturing these organisms and the in progressive destruction of the periodontal ligament and variety of appearances that they present on primary alveolar bone, with pocket formation, recession, or both1. detection, such strains have caused major diagnostic 6 Destruction of periodontal caused by the host defence. difficulties. Clinically periodontitis is characterized by plaque The condition of normal flora is where microorganisms that accumulation, calculus and pocket formation, periodontal are usually in certain locations and under normal tissue inflammation and alveolar bone loss, and gum circumstances do not cause disease6. Granulicatella adiacens bleeding. Pathogenic bacteria are suspected of causing an in the form of small Gram-positive, non-motile, non-spore inflammatory response, gingival and periodontal damage2. coccus, oxidase, and catalase-negative, facultative anaerobes. Periodontal disease can be treated primarily with The nutritional requirements of Granulicatella adiacensare debridement supragingiva and subgingiva. The combination complex and require pyridoxal or L-cysteine in growth of surgical or mechanical treatments and proper oral media as essential nutrition for the normal growth of this hygiene can maintain a healthy periodontal condition3. bacteria7. Granulicatella adiacenshas a normal commensal Periodontal disease is no longer solely caused by the relationship on most mucosal surfaces which allows it to presence of specific bacteria but by ecological changes in the affect the mucosal area of the human body but although biofilm community and the population level of bacterial these bacteria can infect all these areas, these bacteria rarely species in the oral microbiota4. Bacteria in the phylum cause disease. species of this genus found in the oral cavity, firmicutes are lactic acid bacteria, which is commensal intestine and urinary tract8. Because of complex nutritional bacteria in the oral cavity. However, due to a change from growth requirements, Granulicatella adiacenscan most often symbiosis to dysbiosis, this bacteria becomes a pathogen. cause infections in nutrient-rich tissue, such as the Lactic acid bacteria have the ability to ferment sugar into endocardium or the eye, causing endocarditis or lactic acid which can further cause demineralization and endophthalmitis, in addition to causing intravascular bone resorption.5Nutritionally variant streptococcus (NVS) infections. Infection in this area causes endovascular were originally described by Frenkel and Hirsch (1961) as a infections, the central nervous system, ocular, jawbones and new type of streptococcus exhibiting satellitism around joints, and joint and urinary tract infections9. 396 Systematic Review Pharmacy Vol 11, Issue 4, 2020 Harun Achmad et al / Granulicatella adiacens Bacteria Isolation from Perodontitical Patients with Polymerase Chain Reaction Techniques Granulicatella infection is quite alarming for several onto the swab, then the swab put into sterile transport reasons. First, this infection, especially endocarditis, has media. been associated with greater morbidity and mortality compared with other infections caused by another Microbiological Analysis: The swab is inserted into a tube streptococcus. Second, Granulicatella infection has containing the BHIB (Brain Heart Infusion Broth) medium antibiotic resistance and the recurrence rate is higher than then incubated for at least 8 hours at 370 C. The bacteria other streptococcus10. The present study was carried out to culture transferred to Blood Agar medium and incubated determine the presence of Granulicaella adiacens bacteria in for 24 hours at 370 C. chronic periodontitis patients from the subgingival plaque of these patients. DNA preparation: The basic principle of DNA extraction is a series of processes to separate DNA from other MATERIAL AND METHODS components such as proteins and others. This extraction This study was approved by the ethical committee of the procedure includes the process of Sample Preparation, Cell Faculty of Dentistry, Hasanuddin University, Makassar, Lysis, DNA Binding, Wash, Elution. The liquid contained in Indonesia. The study included a total of 18 subjects (8 Male the Eppendorf tube is a DNA product that is ready for PCR. and 10 female) undergoing periodontal treatment. Medical histories from the participants were obtained and thorough Running PCR: Cycle 1 for the Pre-Denaturation process, clinical and radiographic examinations of their teeth were Cycle 2 for the process of denaturation, annealing, conducted after written informed consent. Patients with a extension. Cycle 3 for the final extension process. pocket depth of at least 4 mm; aged 35-50 years systemic disease; OHI-S Moderate-severe; patients do not have a Gel Electrophoresis: systemic disease; were inclusion criteria and the exclusion (Positive Control, Negative Control, sample) were inserted criteria were bacteria does not obtain from the sample and bacterial nucleotides do not exist in Gene Bank. DNA ladder into the well near the positive control. Assessment of periodontal status was performed by a single Electrophoresis is turned on and run from a negative charge trained examiner, the status was means of clinical (cathode) to a positive charge (anode) at 100 A and 60 attachment loss (CAL), probing pocket depths (PPD), which minutes. After electrophoresis, the band is formed. The was measured more than four millimeters per tooth by positive result if the band is aligned with the positive using a WHO periodontal probe. Oral hygiene indices such control, continued by DNA Sequence examination to obtain as papillary bleeding index (PBI), plaque index (PI), and the nucleotide sequence which was then performed a gingival index (GI) were also assessed. BLAST (Basic Local Alignment Search Tool) Nucleotide sequence on NCBI Gene Bank data (National Center for Sample Collection: The sample was collected from the Biotechnology Information) gingival area of buccal tooth surfaces using sterile curettes RESULT Figure 1: Electrophoresis