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Effect of Ethanolic extract of Aloe vera gel on certain common clinical

pathogens.

Article · August 2016

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Borneo Journal of Medical Sciences (2016) volume 10, issue 2, pp: 19-25

Effect of Ethanolic extract of Aloe vera gel on certain

common clinical pathogens
Halima Begum1*, Sadia Choudhury Shimmi2, Mahfuza Mazeda Rowshan3,
Sayeda Khanom4
1
Department of Pharmacology, Enam Medical College, Savar, Dhaka, Bangladesh;
2
Department of Biomedical Science and therapeutics, Faculty of Medicine and Health Sciences,
Universiti Malaysia Sabah, Sabah, Malaysia;
3
Department of Pharmacology, Sir Salimullah Medical College and Mitford Hospital, Dhaka,
Bangladesh;
4
Department of Pharmacology, Central International Medical College, Dhaka, Bangladesh.
*
Corresponding author’s e-mail: [email protected], Tel no: +8801951521350,+8801917964899
(Received: June 10, 2015; Accepted: August 21, 2015)

ABSTRACT

Antimicrobial activity of ethanolic extract of Aloe vera were observed against Staphylococcus
aureus, Staphylococcus pyogenes, Staphylococcus saprophyticus, Streptococcus pneumoniae
and Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae . Aloe vera leaf gel
was used for ethanolic extraction.Zones of inhibition in millimeter was used to measure the
antimicrobial effect. Antimicrobial susceptibility test showed that ethanolic extract has growth
inhibitory effects against the tested pathogens. Ethanolic extract of Aloe vera has growth
inhibitory effects against tested pathogen seen by antimicrobial susceptibility testing.

Keywords: Aloe vera, drug resistance, antibacterials.

INTRODUCTION
Untimely death due to infectious disease has become a global concern world-wide1, 2, 3. Due to
the rapid emergence pathogens, the clinical efficacy of many existing antibiotics are being
threatened 4, 5. Throughout the history of herbals are used to treat infectious diseases6, 7. Pure
compounds or as formulated with measured constituents of plant extracts obtained from natural
product, provide tremendous scope for new drug development. Plant origin antimicrobials are
very limited side effects converse to the synthetic drugs8 and have promising therapeutic
possibility to treat many infectious diseases. There is need to discover new antimicrobials with
novel mechanism of action for new and re-appearing infectious diseases9, 10. Present analysis
was carried out toward traditional plants identification that are effective against the common

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Borneo Journal of Medical Sciences (2016) volume 10, issue 2, pp: 19-25

pathogens such as Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa

K.pneumonia, S.pyogenes, S. saprophyticus and S. pneumonia.
Even though these are very serious human pathogens and often associated with nosocomial
infection, medicinal plants effective against all these pathogens put together and a systematic
study thereafter towards purification of bioactive components is still scanty. Pharmaceutical,
cosmetic and food industries are mainly used Aloe vera products11. Aloes grown mainly in the
dry regions of North America, Europe and Asia. Aloe vera (Aloe barbadensis Miller.), in
Liliaceae family, has leaves30-50 cm long and 10 cm broad, 25-35 cm in length bright yellow
tubular flowers arranged in a slender loose spike; stamens frequently project beyond the
perianth tube 12. Aloe vera contains over 200 active compounds and 75 nutrients like vitamins,
minerals, sugars, enzymes, lignin, anthraquinones, salicylic acid, saponins and amino13. It has
been reported that the antimicrobial constituents of organic extracts of Aloe vera gel have
diverse activity against several human clinical pathogens either by inhibiting the growth or
effective killing14, 15. Because of increasing trends of development of antimicrobial resistance,
purpose of present study was to observe the effect of ethanolic extract of Aloe vera counter to
some common clinical pathogens with emphasis on the acceptability of Aloe vera as a natural
medicine against various infections.

MATERIALS AND METHODS

The study was conducted in the Department of Microbiology at Enam Medical College Savar,
Dhaka, during the period of December 2014 to February 2015(3 months) and a total of 105
different bacterial isolates were tested for antibacterial activity of Ethanolic extract of Aloe
vera. Ethical permission (EM22014) was taken.

Preparation of extract
Leaves of Curacao Aloe (Aloe barbadensis or Aloe vera) collected from Savar, Dhaka. The
plant was verified by taxonomist in Department of Botany, Dhaka University. Fresh leaves of
mature and healthy Aloe vera were washed in tap water for 5 minutes and cleaned with sterile
distilled water. Then by using a sterile knife, the leaves were dissected longitudinally and the
colorless aloe gel (parenchymatous tissue) was scraped out without the fibers. Drained gel was
air dried in the oven at 80 ̊ C for 48 hours. Dried gels were ground to obtain powder by using
mortar and pastel and 30g of this powder was soaked into 300ml of ethanol for 4 days for proper
extraction of the active ingredients at room temperature16. This was later filtered by Whatman
filter paper No.1 and the filtrates evaporated to dry the extract using a rotatory evaporator .The
supernatant was stored in refrigerator at 4 ̊ -C after collection. Before being used for
antimicrobial susceptibility testing the extracts were dissolved in sterile water.

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Borneo Journal of Medical Sciences (2016) volume 10, issue 2, pp: 19-25

Antimicrobial susceptibility testing

The test organisms selected were: Staphylococcus aureus, Staphylococcus pyogenes,
Staphylococcus saprophyticus, Staphylococcus pneumonia (Gram-positive) and Escherichia
coli, Pseudomonas aeruginosa, Klebsiella pneumoniae (Gram-negative).The bacterial isolates
were collected from the Department of Microbiology, Enam Medical College. All the isolates
were identified by standard biochemical methods17. At room temperature seven bacterial
cultures were maintained in nutrient agar medium and were sub cultured every two weeks into
newly prepared nutrient agar slants. Sterile agar (at 45̊ C) was poured into sterile Petri dishes.
The plates were allowed to gel for an hour, which had been inoculated with test organisms.
Standardization of inoculums and ethanolic extract of Aloe vera were done using McFarland
Standards No. 0.5(tube) and corresponding American Type Culture Collection (ATCC) strains.
Antimicrobial susceptibility testing of isolates were done by Disc Diffusion Method18 using
sterile filter paper discs (Whatman filter paper No.1) 6 mm in diameter. For bioassay against
bacteria ethanolic extracts were used. Sterile disc with 6mm diameter were loaded with 0 .1ml
of extract and introduced into sterile medium with the test organisms. The plates were incubated
at 37 ̊ C for 24 hours. The results of antibiogram were evaluated by measuring the zone of
inhibition around the disc and expressed in millimeter.

RESULTS
Ethanolic extract of Aloe vera gel showed varying degree of response regarding antibacterial
property against the tested pathogens. The zone of inhibition with ethanolic extracts ranged
from 12.5 – 22 mm. Zone of inhibition was maximum for S. aureus and minimum for E. coli.
Normally the extracts showed less antibacterial activity against Gram- negative in comparison
to Gram - positive bacteria .The maximum zone of inhibition was 12.5 mm for E. coli, 13.87
mm for P. aeruginosa, 13.22 mm for K .pneumoniae, 22 mm for S. aureus, 20 mm for S.
saprophyticus and 15 mm for S. pneumoniae and 18 mm for S .pyogenes (Figure 1).

Figure 1: Results of Zone of Inhibitions against different isolates

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Borneo Journal of Medical Sciences (2016) volume 10, issue 2, pp: 19-25

DISCUSSION

Antibacterial activity of ethanolic extract of Aloe vera was evaluated on the basis of zone of
inhibition (ZIs) by disc diffusion method in the present study. This ethanolic extract showed
different degree of inhibitory effects against the test pathogens. Among the Gram- positive and
Gram- negative pathogens ,ethanolic extract of Aloe vera showed good inhibitory effect on
S.aureus (22 mm) and the findings were similar with the findings of Agarry Oo(18mm),
Lawrence R et al. (15.66 mm) and Ibrahim et al16, 19,20. The extracts also inhibited the growth
of S. saprophyticus 20 mm and 15 mm for S.pneumoniae, S. pyogens (18mm) and the findings
are almost consistent with the findings of another study conducted by Lawrence R et a l(16
mm)19. In the present study of ethanolic extract of Aloe vera exhibited growth inhibitory action
against E. coli (12.5 mm), 13.87mm for P .aeruginosa, 13.22mm for K.pneumoniae. The
findings of Zone of Inhibitions (ZIs) are similar with findings of Lawrence for E coli but less
for P. aeruginosa and K. pneumonia19. Lawless and Allan and Pugh et al screened antimicrobial
activity against different pathogens and observed minimum inhibitory activity against E.coli14,15
where as Alemdar and Agaoglu in their study found no inhibitory effect against E.coli21, due to
presence of additional lipopolysaccharide layer a lower antibacterial activity against Gram-
negative microorganism may be seen. Furthermore, maximum zone of inhibition against E.coli
was 12.5 mm in this study. The finding correlates with the antimicrobial activity of ethanol
observed in this study. Lawrence R et al in their study isolated the antibacterial compounds
from Aloe vera gel extracts and found Pyrocatechol, cinnamic acid, p-coumaric acid and
ascorbic acid. Possible mechanism of antibacterial actions of that compounds are as -
Hydroxylated phenol- pyrocatechol, which is toxic to microorganisms and its presence in Aloe
vera extract22, 23.The sites and hydroxyl group’s number on the phenol are thought to be relative
toxicity to microorganism and increased in hydroxylation results in more toxicity. Cinnamic
acid in Aloe vera also proved by Dukes phytochemical databases acts by inhibiting resting
bacterial glucose uptake and ATP production24, 25.

CONCLUSION
Globally bacterial resistance of antimicrobial agents is an important human health concern.
Hence, there is need to find out new antimicrobial agents to combat this situation. Thus having
auspicious antibacterial activity and less adverse effects Aloe vera could be used as an effective
antibiotic substitute herbal remedy. Hence, we concluded that Aloe vera gel extract has
opposing effect on the growth of different bacterial pathogens. In vivo study is recommended
to determine its mechanism of action, doses and toxicity.

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Borneo Journal of Medical Sciences (2016) volume 10, issue 2, pp: 19-25

ACKNOWLEDGEMENTS
The authors would like to thank Prof. Iftikhar Ahmed, Head Dept of Microbiology, EMC and
all the other teachers and staffs from Department of microbiology for their technical support.

CONFLICT OF INTEREST
The authors declare that they have no competing interests.

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