Journal Club
Dr Asfand khan
PG Oral and Maxillofacial pathology
SBDC
Journal of Oral and
Maxillofacial Pathology
Impact factor: 1.4
SJR: 0.322
Evaluation of antifungal activity of mint, pomegranate and coriander
on fluconazole-resistant Candida glabrata
Authors: Lekshmy jayan, R ramaya, K rajkumar, N priya
DOI:10.4103/jomfp.JOMFP_355_19
Aims and objectives:
• The aim of this study was to evaluate the efficacy and
resistance of fluconazole and to compare the
antifungal effects of Coriandrum sativum, Mentha
piperita and Punica granatum in C.glabrata.
Introduction..
• Self treatment causes more harm than good.
• Self diagnosis and relying on internet for health problems has led to an
unethical use of various pharmacotherapeutic agents.
• Increased use of antimicrobials , steroidal agents has resulted in
immunosuppression and increased development of antimicrobial resistant
organisms.
• Immunosuppression knocks down the harmony of micro flora, enabling these
organisms to thrive.
Continued..
• The most common opportunistic infection is candidiasis, which is
caused by fungal organisms of genus Candida.
• In the recent decade, these organisms have developed resistance to
various antifungal drugs.
• Although these organisms can be present in normal individuals
without causing disease, it can lead to localized or in rare cases
disseminated candidiasis.
Continued..
• The usage of herbal extracts in the treatment of these infections can
provide a cheaper and simpler alternative.
• These phytochemical agents may help in restoring the previous
susceptibility of the organisms to drugs.
• Unlike the synthetic pharmcotherapeutic agents, these natural agents
have less side effects.
• These herbal plants include coriander, mint, pomegranate.
Materials and Methods
• Method to extract herbal component:
1) Suitable sized M.piperita, C.sativum and P.granatum in an
extractor.
2) Add alcohol 3 times the quantity of raw material.
3) Heat under a reflux at temp 80°C and 85°C for 3-4 h.
4) Filter the extract through filter( 10 μm pore size).
5) The marc is extracted three times more, filtering the extract.
6) The filtrate obtained is then stored.
Continued..
• Method to assess growth of microorganisms:
1) The organism is inoculated into a specific culture medium.
2) This Hichrome candida differential agar media is incubated for 24 –
48h.
3) Growth of the organism is indicated by yellowish creamy, smooth
and raised colony detachable from the agar surface.
4) The presence of the organism is confirmed by KOH and periodic
acid-Schiff(PAS) stain.
Continued..
• Methods to assess efficacy and resistance of C. glabarata to fluconazole:
1) Fluconazole antifungal disc is incorporated in the culture medium.
2) Zones of inhibition are checked for after 20-24h of incubation.
3) If insufficient growth is observed then culture will be incubated again
for 48h.
4) The resistant species will fail to show a zone of inhibition even after
48h.
5) These resistant strains are isolated for further study.
Continued..
• Method to asses antifungal efficacy of mint, pomegranate and
coriander on C.glabrata:
1) The fungal strains were procured and inoculated in the specific
culture medium.
2) Standard concentrations(25,50,75,100,125,150,175 and 200μl)of the
herbal extracts were added to the culture and incubated.
3) The antifungal activity of the extracts were checked and estimated
by the zone of inhibition.
Continued..
• Statistical analysis:
1) The obtained raw data did not follow normal distributions, so
nonparametric statistical analysis was done to evaluate the statistical
significance.
2) Pearson's correlation test was done , as study required a comparison
between different groups.
3) The test enabled a comparison of the efficacy of extract groups, among
themselves and with fluconazole against C.glabrata.
Results:
• The herbal extracts failed to produce zone of inhibition from a
concentration of 25μl till 75μl.
• The result showed an increase in the diameter of the zone of
inhibition from a concentration of 100μl till 200μl.
• Coriandrum sativum(coriander) had the largest zone of inhibition and
the lowest minimal inhibitory concentration among the three extracts
studied.
Continued..
• Although fluconazole resistant strains of C.glabrata were used, all the
extracts used in the study showed an inferior antifungal action to the
fungal strain.
• Zone of inhibition was produced by fluconazole at a concentration of
250mg comparable to the herbal extracts at a concentration of 200μl.
• Analysis of the values between positive group and study group
showed P<0.001 in all three extracts, they are statistically significant.
Continued..
• Significant difference between the antifungal properties of fluconazole and
the herbal extracts against resistant C.glabrata.
• Comparing the three different extracts among themselves gives a P value
more than 0.001, they are considered statistically insignificant.
• The antifungal activity of the herbal extracts are almost equally potent.
• Correlation value was close to +1, which means these three extracts have a
positive correlation.
Discussion:
• Antimicrobial resistance is one of the biggest therapeutic challenges
in the management of various infectious disease.
• Injudicious use of antimicrobial agents has led to the development of
resistant microorganisms.
• Phytochemical agents or herbal therapeutic agents can be a safer
alternative, for the management of resistant strains of
microorganisms.
Continued..
• In the current study all the three herbal extracts showed comparable
antifungal properties without any significant difference.
• All the three herbal extracts were significantly superior to fluconazole.
• Msaada et al.(2014) reported that coriander has a potent antioxidant
properties as a product of high concentrations of flavonoids, tannins
and polyphenols.
Continued..
• In the current study the antifungal activity of C.sativum was superior
to that of fluconazole, in fluconazole resistant C.glabrata.
• Coriander showed a clear zone of inhibition around the colonies,
suggesting complete lysis of the organisms .
• Rongai et al (2019) reported that wild genotypes of pomegranate
CREA-FRU6, CREA-FRU11 and CREA-FRU76 have potent antifungal
activity.
Continued..
• In the current study, pomegranate in higher concentration showed a
statistically significant antifungal action, superior to fluconazole.
• The results of this study contradicted the results obtained by Nicole et
al,(2014) who reported that pomegranate extract lacked antifungal
property against Candida species.
• It has been proven that mint possesses anti parasitic, antibacterial,
analgesic and bug repellent properties.
Continued..
• This study showed a greenish zone of inhibition around the colonies,
and mint showed a superior antifungal action to fluconazole.
• The results of the present study are similar to study conducted by
Wenji et al, (2019) , who reported that mint leaves have antifungal
activity against C.albicans at a concentration of 80%.
Limitations:
• Being an in vitro study, the authors were unable to assess the effects
of systemic factors or systemic illness on the mode of action of these
herbal agents.
• It is not known, if these herbal agents may have an antagonistic or
synergistic effect with any other medication administered for different
purposes.
• Another limitation of the present study was the use of crude herbal
extracts.
Continued..
• Furthermore, since it was in vitro study, the
effects of other conditions in the oral cavity
or body, which may act as a confounding
factor in the effectiveness of the drug could
not be evaluated.
Conclusion:
• The need of the hour, is the discovery of newer drugs in treating
infections, especially resistant antifungal infections.
• All the three herbal extracts used in this study showed potent
antifungal activity against C.glabrata, and values were found
statistically significant.
• These agents can be safer and cheaper alternative, to the present
therapeutic systems, without any harmful side effects.
Continued..
• These herbal extracts may be used alone or can be used as a
multidrug combination with fluconazole.
• This will reduce the dosage needed for the management of the
infectious conditions.
• Further studies are needed to see, if their efficacy is impeded by any
systemic or local factors.
Thank you
• Drug interactions of herbal extracts with other medications should
also be evaluated.
• As this infection is more prevalent in patients who are suffering from
diabetes, immunosuppression and acquired immunodeficiency
syndrome.
