3314 Indian Journal of Forensic Medicine & Toxicology, April-June 2021, Vol. 15, No.

Antimicrobial Activity of Lactic Acid Bacteria Found in

Dadiah on Disease-Causing Skin Infections

Rinita Amelia1, Dessy Abdullah2, Yudha E. Pratama3, Endang Purwati4

1,2
Doctoral Program Student Lincoln of University College, Faculty of Health and Science, Bharu City Malaysia,
1,2
Lecture Baiturrahmah University, Medical Faculty, Padang Indonesia, 3Doctoral Program Student Andalas
University, Faculty of Animal Science, Padang, Indonesia, 4Professor Andalas University, Faculty of Animal
Science, Biotechnology/Technology of Product Husbandry, Padang Indonesia

Abstract
Dadiah is a fermented food made from buffalo milk containing lactic acid bacteria (LAB). The potential
of LAB as an antimicrobial organism is expected to inhibit or kill pathogens, especially those causing
inflammations in the skin. Dadiah used in this study was obtained from the Lintau area of Tanah Datar
District, West Sumatra, Indonesia. LAB isolation and identification involved the use of MRS Broth
(MERCR) and MRS Agar (MERCK) media and the pathogenic bacteria used include Staphylococcus
aureus, Propionibacterium acnes, Acinetobacter Baumannii, Klebsiella pneumonia, and Pseudomonas
aeruginosa. The results of this study obtained a total LAB of 7x1011 CFU / g and showed the highest
antimicrobial activity in the Staphylococcus aureus with a14.53 mm clear zone and 13.18 mm bacteria in
the Pseudomonas aeruginosa test bacteria.

Keywords: Antimicrobial dadiah, lactic acid bacteria, skin diseases

Introduction viable solutions in form of alternative therapy or

Skin diseases are the most common types of diseases treatments[1]. These forms of alternative therapy include
in developing countries, especially in those with tropical synthetic antibiotics.
climate. Therefore, humid and hot air throughout Probiotics, vitamins, and phenolics are a few
the year is highly suitable for the development of examples of nutraceuticals. These substances are
infectious, disease-causing pathogens in the skin. The important for skin protection and health, and they are
prevalence of such infections in developing countries known to assist in medical treatment of dermatological
ranges between 20-80%. As a developing country, problems[2]. This study aims to discover the role of
Indonesia and its inhabitants are prone to skin diseases probiotics (Lactobacillus) as a prevention and treatment
of which its main causes include poor geographical and for skin infections. Many studies have shown that lactic
hygiene conditions. Such conditions enhance the rapid acid bacteria (LAB) helps to activate commensal bacteria
development of pathogens and bacteria. Therefore, the due to its probiotic properties, and gets rid of pathogenic
attention and recommendations of appropriate health bacteria through the antimicrobial properties derived
organizations in the preventive measures against skin from either bacteriocin found in LAB itself, or the role
infections is presently a necessity. The problem of of its peptides. Some studies also mention that the role
antibiotic resistance has become a worldwide problem, of probiotics in nutrition involves boosting the immunity
especially during prevention and cure, that necessitates against skin infections caused by various pathogenic
bacteria such as Aeromonas bestiarum, Ichthyophthirius
multifiliis, Staphylococcus aureus, Corynebacterium
Corresponding Author:
acnes, Streptococcus, etc[3].
Prof. drh. Hj. Endang Purwati MS, PhD
[email protected] / [email protected]
 Indian Journal of Forensic Medicine & Toxicology, April-June 2021, Vol. 15, No. 2 3315

Dadiah is a traditional food from West Sumatra made in an anaerobic jar and incubated for 48 hours with a
of buffalo milk that undergoes a natural fermentation temperature 37oC. After incubation, a single colony of
process in a bamboo tube, Dadiah originally made LAB was obtained and hence isolated [10].
from Lintau in Tanah Datar, contains probiotics such
Identification of Lactic Acid Bacteria
as L. Plantarum which has proven to be effective in
maintaining the balance of intestinal microflora, and Identification of LAB from Dadiah samples was
contains strong antimicrobial properties[4]. done by placing 1g of the sample in a test tube containing
9 ml MRS Broth diluted up to ten (10-1) logarithmic
Atopic dermatitis is an inflammatory skin disease
fold. It was further incubated in anaerobic conditions for
occurring often in children and the elderly [5]. The
24 hours, after which a 100 uL 10-1 dilution was added
dominant bacterial species found in this skin disease is
to the tube containing 900 uL 1 MRS broth dilution 10-
Staphylococcus aureus. The colonies of this bacteria,
2, and a subsequent dilution of 10-8. Consecutively, 100
when present in the skin, can result in a wide range of
uL of the serial dilution of 10-8 was inoculated on the
infections therein. Therefore, antimicrobial treatment is
media MRS Agar with the method of spread, placed
necessary to prevent such bacterial growth [6] . Many
within an anaerobic jar, and incubated for 48 hours at
broad-spectrum antibiotics can be used as an inhibitor of
37 0C. A single (choice random) colony that is round,
Staphylococcus aureus, such as penicillin, doxycycline,
slippery, and either white or yellowish is characteristic
tetracycline, clindamycin, etc. However, improper
of LAB. Finally, this bacterial colony was transferred
use of an antibiotic results in drug resistance. It is
via streak method and incubated [10].
also associated with an increased risk of infection and
emergence of bacterial colonies, known as “Multidrug- Antimicrobial Activity
Resistant Organisms (MDRO)” [7]. These Methicillin
resistant Staphylococcus aureus (MRSA) and MDRO The antimicrobial resistance testing was carried
are the main causes of nosocomial pathogenic infection out using five bacteria samples that cause skin disease.
worldwide, and require serious treatment, as they LAB culture (1 mL) was centrifuged at 10,000 rpm for
increase comorbidity and death due to the infection[8]. 5 minutes at 270 C, and supernatant volume of 50 ᶙl was
A major challenge posed by S. aureus is antimicrobial used to test for antimicrobial resistance. 0.2% of test
resistance. After the introduction of the antibiotics bacteria (Staphylococcus aureus, Propionibacterium
penicillin and methicillin into clinical practice, strains of acnes, Acinetobacter Baumannii, Klebsiella pneumonia,
antibiotic-resistant S. aureus (MRSA) were subsequently and Pseudomonas aeruginosa) was placed on Nutrient
identified [9]. Agar (NA) as a growth media for test bacteria,
homogenized, injected by micropipette, and incubated at
Therefore, this study was aimed to determine 37 0C. Clear and circular inhibition zones are measured
the role of Dadiah as a probiotic that can be used to after 12 hours [11].
inhibit bacteria-induced skin diseases, especially the
inflammatory infections, as a supporting or future Results and Discussion
treatment alternative against skin diseases.
Isolation Lactic Acid Bacteria
Materials and Methods Dadiah originating from the Lintau area of Tanah
Isolation of Lactic Acid Bacteria Datar District, West Sumatra, Indonesia is known to
contain Lactic Acid Bacteria (LAB). It was isolated with
One gram of Dadiah sample was placed within test MRS as a specific media for LAB growth and incubated
tubes containing 9 ml of MRS broth and diluted up to for 48 hours. In this study, the amount of LAB used
ten logarithmic (10- 10) fold. Furthermore, it was added was 7x1011 CFU / g. This result is higher compared
to test tubes containing 900 ul of MRS broth variations to those obtained, while the total LAB obtained from
of dilution ratio 10-1, 10-2 and 10-7. Furthermore, 100 Payakumbuh Dadiah was 8x108 CFU/g [12], in a research
ul of the serial dilution of 10-9 was inoculated into at Solok district, the total LAB obtained was as much
the MRS Agar media using the spread method, placed as 8x108 CFU/g [13]. This is because Dadiah kicks off
3316 Indian Journal of Forensic Medicine & Toxicology, April-June 2021, Vol. 15, No. 2

its fermentation spontaneously; without the need for a that the bacteria were Gram-positive and rod-shaped.
fermentation starter. LAB was also found in buffalo milk Lactic acid bacteria (LAB), belongs to a group of gram-
obtained from several areas in West Sumatra totaling positive, catalase-negative bacteria, which synthesize
3x106 - 2x108 CFU/g [14]. This is consistent with the lactic acid by the fermentation of carbohydrates. The
standards which state that LAB in probiotic food must cells are known as coccus or bacillus, usually arranged
be present within the range of 106 - 108 CFU / Gram [15]. in pairs or chains. LAB are facultative anaerobes, non-
motile and mesophyllic [16].
Identification of Lactic Acid Bacteria
Antimicrobial Activity
Incubation was done using MRS Broth and
then cultured on MRS media. For 48 hours in 370 Antimicrobial testing using Staphylococcus aureus,
C incubators, a single colony was obtained through Propionibacterium acnes, Acinetobacter Baumanni,
macroscopic observations with round, cream-white Klebsiella pneumoniae and Pseudomonas aeruginosa
and convex shapes. Microscopic observations showed commonly found in skin infections can be seen in Table
1.

Table 1. Diameter Clear Zone

Bacteria test (mm)

Isolate
Staphylococcus Acinetobacter Klebsiella Pseudomonas
Propionibacterium acnes
aureus ATCC baumannii pneumoniae aeruginosa

3A 14,53 7,6 10,24 10,15 13,18

Testing antimicrobial activity involved a well- with Lactobacillus casei strain of Shirota bacteria of 11.2
known method of random selection of 3A isolates from mm and therefore higher in inhibitory power than the L.
curd which have the potential to function as probiotics, acidophilus HM1 bacteria of 3.3 mm [18]. The clear zone
because they can inhibit the growth of pathogenic of Acinetobacter baumannii (10.24 mm) obtained in this
bacteria which cause skin diseases. The clear zone study is greater than that of which used the Lactobacillus
produced by lactic acid in table 1 shows the existence of acidophilus LMEM8 against Acinetobacter baumannii
antimicrobial activity from LAB isolated from Dadiah and obtained a diameter of 5.13 mm [19]. The results of
samples obtained from Lintau, Tanah Datar District. The this study are more productive than that of in testing
ability of LAB to inhibit pathogenic bacteria is due to the the antimicrobial activity of Lactobacillus fermentum
presence of its metabolites such as lactic acid, peroxide, MTCC 9748 against Klebsiella pneumoniae NTCC
and bacteriocin. 703603 which resulted in a 9.17 mm clear zone [20].
Moreover, the Pseudomonas aeruhginosa test bacteria
Clear zones with the highest numbers were produced
(13.18 mm clear zone) was not significantly different
in the Staphylococcus aureus ATCC test bacteria (14.53
compared to the research testing of Lactobacillus spp
mm) from Dadiah isolates. This clear zone is smaller
against Pseudomonas aeruginosa bacteria with a range
when compared to the study of which used LAB
of clear zones 11-21 mm [21].
isolated from tempoyak which inhibits the bacterium
Staphylococcus aureus which a diameter of 19.3 mm [17]. Conclusion
In Propionibacterium acnes, test bacteria obtained a Lactic acid bacteria isolated from Dadiah of Lintau
clear zone of 7.6mm, the smallest diameter compared to origin is probiotic in nature, and it shows a highly
other test bacteria. This result is smaller when compared significant inhibitory effect against Staphylococcus
 Indian Journal of Forensic Medicine & Toxicology, April-June 2021, Vol. 15, No. 2 3317

aureus (of which the largest clear zone of 14.53mm Journal of Antimicrobial Agents Role of probiotics
was obtained), Propionibacterium acnes, Acinetobacter in the prevention and treatment of meticillin-
baumannii, Klebsiella pneumoniae and Pseudomonas resistant Staphylococcus aureus infections.
aeruginosa. However, further research is necessary International Journal of Antimicrobial Agents,
in order to ascertain other probiotic, antimicrobial 2020; 42(6): 475–481.
candidates which inhibit pathogenic bacteria which 9. Creech, C. B., Al-zubeidi, D. N., & Fritz, S. A.
cause skin diseases and infections. Prevention of Recurrent Staphylococcal Skin
Infections, 2015; Vol 29: 429–464.
Conflict of Interest : None 10. Purwati, E., S. Syukur, dan Z. Hidayat. Lactobacillus
Ethical Clearance: Not required in an in-vitro sp. Isolasi dari Bivicophitomega sebagai Probiotik.
Di dalam Proceeding Lembaga Ilmu Pengetahuan
studys
Indonesia, Jakarta. 2005.
Source of Funding : Self 11. Melia, S., Purwati, Yuherman, Jaswandi,
Aritonang, S. N., Purwanto, H. Supernatan Bebas
Acknowledgement: Author Acknowledge the great Sel Lactobacillus fermentum L23 yang berpotensi
help received from the laboratory microbiology and sebagai Antimikroba. No. Paten IDS000002471.
laboratory technology of animal product, Faculty of Dirjen Kekayaan Intelektual, Kementerian Hukumn
Animal Science, Andalas University and special thanks dan HAM. 2019.
for Prof. Endang Purwati PhD. 12. Syukur, S., Yolanda, S., and Fachrial, E. Isolation ,
antimicrobial activity and bioremediation of heavy
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