1.

Title : Further Investigation of Piper crocatum Effect as Anti-Bacterial

Agent to Resistance Propionibacterium acnes
2. Background
Acne is the most common human skin disorder with a very high
prevalence. Antibiotic therapy directed against Propionibacterium acnes has been
a mainstay of treatment for more than 40 years. Despite years of widespread use
of systemic tetracyclines, erythromycin, and clindamycin change in P. acnes
sensitivity to antibiotics was not seen until the early 1980s and became a
worldwide problem in this recent years.
Widespread use of tetracyclines and erythromycin occurred for more than
25 years before less-sensitive strains and clinically relevant or resistant strains
were identified. In the late 1970s, a few strains of P. acnes that were relatively
insensitive to erythromycin and clindamycin were first reported and were not
viewed to be clinically significant [1].
Nowadays, most of people in Indonesia tend to consider the use of herbal
medicine from the nature. The advantages of herbal medicine become an
interesting discussion. Most information expand in society is empirical rather than
scientific evidence. Thus red betel vine's benefit also becomes interesting
discussion but evidence based medicine about it is less known.
Since the treatment to heal the acnes symptoms mostly used the antibiotic
which come from Ascomycetes class of fungus and show the effect to resistance-
bacteria, the other sources of anti-bacterial agent is need to be explored.
The result in previous study shows that the ethanol extract of Piper
crocatum is potentially inhibit the growth and degenerate the Staphylococcus
aurea ATCC 25923 (Gram-positive bacteria) in concentration 25% [2]. Since the
P.acnes is included into gram-positive bacteria, further investigation is needed to
be experimented to know the effect of the extract to P.acnes.
3. Objectives
To test the effect of ethanol extract of Piper crocatum as anti-bacterial
agent to resistant Propionibacterium acnes.
4. Literature Review
Piper crocatum Ruiz and Pav. is traditionally used by Indonesians for
treating various diseases [3]. Red betel (Piper crocatum) is one of the medicinal
plants empirically known potential has healing properties various types of
diseases. Efficacy and benefits of red betel contained therein including, Flavonoid
compounds and Polivenol, Alkaloid, Tanin, and atsiri oil. These compounds are
known have function as antibacterial.
The compounds in red betel can also be used as a barrier to the growth of
cancer cells. Karvakol content of the red betel leaves useful as disinfectant, and
anti-fungal, thus serving as a mouthwash and discharge medications. Eugenol-
containing compounds serve as a pain reliever or analgesic. The content of tannins
serve as healers especially diarrhea and abdominal pain can also be used as an
antiseptic on the wound.
Flavonoids function as Antibacterial by forming complex compounds
against protein which interfere with the integrity of extracellular bacteria cell
membrane. Flavonoids are phenolic compounds as phenol compounds can be
coagulator protein. Alkaloids have the ability as an antibacterial. Mechanism is
suspected by interfering components of peptidoglycan in bacterial cell, so the cell
wall lining not fully formed and cause the cell death. Tannins have activity
antibacterial, an outline mechanism is expected as follows: toxicity tannins can
damage the bacterial cell membrane, compound astringent tannins may induce
formation of complex compounds bond against microbial and enzyme or substrate
formation of a complex bond tannins to the metal ion tannins add to the toxicity.
Tannin is believed to constrict cell wall or cell membrane so that interfere
with the cell permeability itself. Due to disruption permeability, cells cannot
perform living activities so stunted growth or even dead. Masduki states that
tannins also has antibacterial power protein precipitate manner, tannins for
allegedly having an effect The same with phenolic compounds. Effect
antibacterial tannins, among others through: reaction with cell membranes,
inactivation enzyme, and the destruction or inactivation function of genetic
material.
Essential oils act as Antibacterial by interfering membrane formation
process or cell wall thus not formed or not perfectly formed. Essential oils active
as antibacterial generally contain functional groups hydroxyl (-OH) and carbonyl.
Derivative phenols interact with the bacterial cell through a process involving
adsorption hydrogen bonds. At low levels of phenol-protein complex formed with
weak bonds and immediately undergo decomposition, followed by penetration
phenol into the cell and cause precipitation and denaturation of proteins. On high
levels of phenols cause coagulation proteins and cell membranes experienced lisis.
Propionibacterium is a Gram-positive, rod shaped genus
of bacteria named for their unique metabolism: They are able to
synthesize propionic acid by using unusual trans-carboxylase enzymes Its
members are primarily facultative parasites and commensalisms of humans and
other animals, living in and around the sweat glands, sebaceous glands, and other
areas of the skin. They are virtually ubiquitous and do not cause problems for
most people, but Propionobacteria have been implicated in acne and other skin
conditions. One study found that Propionibacterium were the most prevalent
human skin-associated microorganism .P acnes grows in the lipid-rich
microenvironment of the hair follicle. In acne vulgaris, P acnes produces
inflammatory mediators that result in acne papules, pustules, and nodulocystic
lesions [4]. .
5. Research Methodology
Study includes sample preparation, making extracts, and test the
antibacterial resistance. Test material used is an extract of red betel (Piper
crocatum).
The steps to make the extraction of red betel leaf (Piper crocatum)
First, the leaf of red betel is washed to be clean. Thus, cut the red betel
leaf (Piper crocatum) so that it becomes a smaller part. After that, the is
died by using an oven at a temperature of 50 2 C for two days. Red
betel leaf which has been dried milled with a blender to produce a smooth
material. Thus, Soxhlet apparatus is prepared for extracting. Enter 96%
ethanol in a round bottom flask was soxhlet money ( 500 ml). Enter the
red betel leaf which has been refined in the Soxhlet flask that had been
given a paper filter ( 500 grams). Results obtained extract was then
evaporated the solvent with electro-manthel at 60 C until all the solvent
is gone. Filter results extraction with filter paper and insert it into the
extraction bottle. The extraction is ready-made in 100%level [5].
The antibacterial test capability used liquid serial dilution method. The
serial dilutions of Piper crocatum: 100%, 50%, 25%, 12.5%, 6.25%,
3.13% and 1.56% thus bacterial suspension is added in the same volume
[6]. There are 4 controls the media controls, the extracts control, bacteria
control and antibiotic control. Media with the lowest concentration that
remains clear is used for MIC ( Minimum Inhibitory Concentration) for
the test material. Furthermore all scratched on Blood Agar media / Mac
Conkey to know its MBC (Kill Minimal Levels). The Blood Agar Media
is appropriate to P.acnes growth [7].
6. Reference
Eady et al. 2003. Propionibacterium acnes Resistance: A Worldwide Problem.
Dermatology Journal 206:5456
Juliantina et al. 2009. Manfaat Sirih Merah (Piper Crocatum) Sebagai Agen Anti
Bakterial Terhadap Bakteri Gram Positif Dan Gram Negatif. Jurnal
Kedokteran dan Kesehatan Indonesia
Wicaksono et al. 2009. Antiproliferative Effect of the Methanol Extract of Piper
crocatum Ruiz & Pav Leaves on Human Breast (T47D) Cells In-vitro.
Journal of Pharmaceutical Research 8 (4): 345-352
http://en.wikipedia.org/wiki/Propionibacterium_acnes
Oktaviani, Dina. 2012. Uji Banding Efektivitas Ekstrak Daun Sirih Merah (Piper
crocatum) Dengan Zinc Pyrithione 1% Terhadap Pertumbuhan
Pityrosporum ovale Pada Penderita Berketombe. Karya Tulis Ilmiah. FK
UNDIP
Zandi et al. 2011. Determination of microbial agents of acne vulgaris and
Propionibacterium acnes antibiotic resistance in patients referred to
dermatology clinics in Kerman, Iran. Jundishapur J Microbiol. 2011;
4(1): 17-22.
Propanidisa Mikro and Molecular Biology. 2003. Microbiology Culture Media
Manual. Available at www.condalab.com