I gratefully acknowledge and appreciate her help in preparing the clinical trials, and teaching us at the beginning of the project. To my internship partner Tiara, I am grateful for her help during all the trials and tribulations during this project and for always being encouraging and having a cheerful attitude, and to my colleagues Fanie, Fenny, Aurelly and Sonya, I humbly thank them for all their hard work and dedication to make this project successful despite all the minor setbacks that occurred. This internship report deals with the "Antibacterial evaluation of deodorant X and bacterial identification of armpit in young adult men via clinical trials", which is done in implementation of the internship course, which was held from June to August 2022.
Deodorant is a personal hygiene product that has the function of combating body odor through many mechanisms, one of which is inhibiting the growth of bacteria. Thus, the results can conclude that although the antibacterial effect of the deodorant is not significant, it reduced the growth of bacteria and some bacteria identified were the species responsible for the production of body odor. Their vision is to "become a 'leading interdisciplinary institution of higher education that is globally connected and directly impacts society through science and innovation.'" i3L provides many state-of-the-art laboratory facilities and equipment for their many laboratories.
Skinproof focuses on beauty, health and wellness, with an emphasis on research and testing of cosmetic and personal care products that promote the overall health of the human body. The i3L product is a Skinproof project, which is a multidisciplinary research project approved and funded by Skinproof. In addition, the Skinproof project also conducted an in vitro study to test several antimicrobial activities of deodorants.
This practice report details part of the Skinproof project, which is one of the clinical trials.
PROJECT DESCRIPTION
Internship Project
- Project background
- Scope of the project
- Objectives / Aims
- Problem formulation and Proposed Solutions
- Clinical Trial Preparation .1 Pre-clinical Trial Preparation
- Clinical Trial
- Bacterial Identification .1 Preparation of MSA Agar
- Statistical Analysis
The scope of what is involved in a clinical trial, from preliminary preparation to interpretation of results, includes essential documentation preparation, subject selection, material preparation, sample collection and an antibacterial study, all of which are carried out as part of the Skinproof project. This particular clinical trial is part of a larger project that has been conducting clinical trials for other deodorants with the same goal of evaluating the antibacterial properties of the deodorant. Each of the participants must read and sign a consent form to obtain informed consent for the trial, and the ethics permission document must be approved by the Ethics Committee of Fakultas Kedokteran Universitas Indonesia (FKUI) to proceed with the study. .
For the conditioning period, the subjects were required to: shave the axillary hair on the first day of the conditioning period and avoid the hair for the rest of the conditioning period, avoid the use of any axillary dermatological product, avoid heavy activity, spicy avoid food, and use only the given soap. The agar used to evaluate the antibacterial activity of the deodorant is Mueller Hinton agar (MHA), which is commonly used in antibiotic susceptibility testing (AST) because it is optimal for the cultivation of non-stringent bacteria, and supports reproducibility for multiple groups of tests. , a reason relevant to the clinical trial (Hudzicki, 2009). The solidified and paraffin wax-sealed agar plates were stored in the refrigerator at 4 °C, or used immediately.
Phosphate-buffered saline (PBS) was used to store the bacterial swab sample. The now cooled solution was then aliquoted into microcentrifuge tubes to be used for serial dilution during the microbial study. Miles and Misra was chosen for the microbial study due to the fact that it can be widely used to determine the number of viable bacterial colonies, that it is inexpensive, and that one plate can contain an abundance of colony-forming droplets to be counted, and thus saving time and agar plates (Hoben & Somasegaran, 1982).
Before plating, the agar plate was divided equally into 6 sections for the five different dilutions and the control, and 10 µL of the dilutions created as well as the control (PBS only) were then, using a micropipette, in triplicates in their respective pipetted pipetted sections (Figure 3). After the drops were pipetted, the agar plate in the BSC was left to dry, then sealed with paraffin wax to prevent contamination, and incubated for 24 h at 37 °C. In the case of this study, knowing which species of bacteria typically colonize the armpit area allows for the correct determination of whether or not the deodorant is able to inhibit the bacteria responsible for body odor.
Like MHA agar, MSA agar was prepared in the same way according to the manufacturer's instructions. The medium is differential due to the presence of mannitol in the medium as it is used as a substrate for fermentation and the medium is considered selective due to its high concentration of NaCl which allows the growth of Staphylococcus spp while inhibiting the growth of all other bacteria. When fermentation takes place in the agar with the help of bacteria, the color of the pH indicator, phenol will be red, yellow.
The spread sample allows a diluted sample to be spread over the surface of the agar, enough so that individual colonies can grow far enough apart to be taken up by an inoculation loop (Leboffe & Pierce, 2015). In the study, 50 µL of the stock sample was pipetted onto the agar and spread uniformly over the agar with a glass rod.
FINDINGS
Result
- Antibacterial Study
- Bacterial Identification
The percent reduction was also calculated to determine the reduction in LogCFU/mL values from the control axilla, representing the pre-treatment value, and the treated axilla, representing the post-treatment value (see Table B1). MSA was used to identify some bacteria taken from the axilla of young adult male participants. Macroscopic observations of all spread plates were made after 24 hours of incubation at 37 °C, in addition to the Gram method (see Figure C1).
Microscopic observations of bacteria were made using a light microscope, at 100x magnification, and Gram staining was done to stain bacterial cells. As shown in Figure 6A, after incubation, the agar appeared yellow and the morphology of the bacterial colonies was yellow in color, circular in shape, appeared smooth and shiny, and convex in height. Microscopic view, at 100x magnification after Gram staining, the bacterial cells appeared to be heads, attached together in clusters, and the color of the cells after Gram staining was purple (Figure 6B).
There was also macroscopic observation of another agar plate that showed a single yellow circular convex colony, the zone around the colony was yellow while the rest of the agar remained pink (Figure 6C). Finally, Figure 6E shows white, circular, convex colonies without any yellow zone surrounding them, and Figure 6F shows microscopic observation after Gram staining of the white colonies, revealing purple cells in clusters.
Result Interpretation and Discussion
For bacterial identification, Gram staining was performed, in which the classification of the bacteria based on their cell wall components could then be determined, further allowing for a better prediction of what the bacteria might be. The selective and differential media used was MSA, which is used to isolate Staphylococcus spp. to isolate, and the reason why only these media were used is because the bacteria of interest that frequently colonize the axilla and cause body odor, Staphylococcus spp. is. Macroscopic observations of the MSA after incubation of the dispersed bacterial sample show that the agar has changed color from pink to yellow, which according to Leboffe & Pierce (2015) indicates that the bacterium is most likely S.
Based on microscopic observations of the morphology of the bacteria after being subjected to gram staining, the bacteria are gram-positive, and it can be concluded that one of the bacterial species isolated from the armpit of one of the patients is S. This type of staphylococcus is known to ferment mannitol. and is known to be gram-positive (Leboffe & Pierce, 2015). The same can be said for the bacterial cells obtained from the agar shown in Figure 6C, as there is a yellow zone surrounding the yellow colony, which indicates that mannitol fermentation has occurred, and after Gram staining and microscopic observation, the bacteria is found to be gram-positive, indicating that it is S.
The identity of the bacteria on the agar plate shown in Figure 6E, however, may be a different Staphylococcus species than S. This is suggested by the absence of yellow areas or halos surrounding the bacterial colonies and by the white color of the colonies. Bacterial cells were observed to be purple, indicating that they are gram-positive and appear to be Staphylococcus, based on their clustered and circular appearance.
CONCLUSION AND RECOMMENDATION
SELF REFLECTION
Antibacterial activity of shikimic acid from pine needles of Cedrus deodara against Staphylococcus aureus through cell membrane damage.
