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Effectiveness of Neem (Azadirachta indica A. Juss) Oil against Decay Fungi

Article · January 2017

DOI: 10.22232/stj.2017.05.01.06

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 Science and Technology Journal, Vol. 5 Issue: I ISSN: 2321-3388

Effectiveness of Neem (Azadirachta indica A. Juss)

Oil against Decay Fungi
Kanchan Rawat1*, Uttam Kumar Sahoo2, Nagaraj Hegde3 and Awadhesh Kumar4
1,2,3
Department of Forestry, Mizoram University, Aizawl, Mizoram–796004
4
Department of HAMP, Mizoram University, Aizawl, Mizoram–796004
E-mail: *kanchanfri@gmail.com

Abstract—The enormous use of metallic wood preservatives has caused destructive impact on environment as well
as human health. Therefore realizing the urgency of switching to Environment friendly options such as natural oils are
being tested for their antimicrobial properties. The present study aimed at investigating potential of Neem oil against the
growth ofdecaying fungi. The ability of Neem oil to inhibit mycelia growth of Schizophyllum commune, Fusarium oxysporum,
Fusarium proliferatum, Coniophora puteana and Alternaria alternata was tested at different concentrations of 0.25, 0.50,
0.75, 1.0, 2.0, 4.0, 6.0, 8.0 and 10%. Results of the study revealed Neem oil concentrations above 2% were signi icantly
inhibitory to all the tested fungi.

Keywords: Antimicrobial, Neem Oil, Decaying Fungi, Mycelia, Wood Preservatives

INTRODUCTION MATERIALS AND METHOD

Bamboo is the best renewable natural resource with more
than 1500 uses. Its strength comparable to timber, diversity
and abundance in tropical and subtropical parts of world
has made it a potential replacement for steel and timber
(Scurlock et al. 2000). But the major drawback in using
bamboo is its low natural durability. Bamboo has high
amount of starch and negligible amount of extractives unlike
timber, therefore, it becomes an easy prey to fungi (Kumar
et al. 1994; Wei et al. 2013). With preservative treatment
the service life of bamboo can be effectively enhanced but
unbridled use of chemicals in wood preservation industry
has brought human as well animal population at the verge
of health threat. In many countries some of carcinogenic
preservatives have been already banned (Edlich et al. 2005).
Therefore, to explore antimicrobial potential of natural
plants scientists have carried out extensive research on
extracts and oils of various plant products. In present study
Neem oil has been tested against various fungi including
white rot, brown rot, soft rot and sap stain which cause
decay in wood as well as bamboo (Sharia et al. 2012; Kumar
et al. 2013; Hyde et al. 2001; Schmidt et al. 2011). Neem
oil is known to possess antimicrobial propertied and has
been widely studied for its potential against fungi
(Khetarpal 2010).
http://doi.org/10.22232/stj.2017.05.01.06
Neem seed oil/ Neem oil was procured from themarket
with brand name VyasNeem tail.Fungal strains of bown
rot Coniophora puteana (Acc. No. MTCC 1068), white rot
Schizophyllum commune (Acc. No. MTCC 1096), sap stain
Alternaria alternata (Acc. No. 2060) were obtained from
The Microbial Type Culture Collection and Gene Bank
(MTCC), IMTECH Chandigarh and was maintained on
Potato Dextrose Agar (PDA) medium at 25 ± 2°C. The soft
rots Fusarium proliferatum and Fusarium oxysporum were
provided by The Department of Horticulture, Aromatic and
Medicinal Plants. These strains were also maintained on
Potato Dextrose Agar medium at 25 ± 2°C.
OISONED FOOD TECHNIUE ȍFȎ
The antifungal property of Neem oil against the test fungi
was assessed using the Poisoned Food (PF) technique.
Different concentrations of Neemoil (0.25, 0.50, 0.75, 1.0,
2.0, 4.0, 6.0, 8.0, 10.0%) were added into the potato dextrose
agar medium and 30 ml of medium was poured into each
sterilized Petri dish (9 mm diameter). A inal concentration
of 0.5% (v/v) DMSO was used to dissolve Neem oil in PDA.
DMSO was used as positive control. After inoculation with
5mm diameter mycelia discs were cut from the periphery of
a freshly growing PDA culture, plates were incubated for 15
 Effectiveness of Neem (Azadirachta indica A. Juss) Oil

days at 25 ± 1°C and 70 ± 2% relative humidity. Five replicates
were taken for each concentration. The lowest concentration
which inhibited the visible growth of the fungus was
considered as minimum inhibitory concentration (MIC).
Percentage of mycelia growth inhibition was calculated by
the following formula:
Where,
‘dc’ is the average diameter of fungal colony at 0.0%
concentration of Neem oil,
‘dt’- denotes the average diameter of fungal colony in the
treatment.
RESULTS AND DISCUSSION
Neem oil has proven ef icacious as an insecticide. The
present study is an attempt to evaluate its ef icacy as
a potential bio fungicide. The results of fungal growth
inhibition at different concentrations of Neem oil (0–10.0
%) are shown in Fig. 1.
At same concentration F. oxysporum and A. alternata were
inhibited to fairly good extent as 16% and 12% respectively.
At 0.0% concentration there was 100% growth of all the
tested fungi. While in positive control (DMSO) there was
2.20, 5.00, 2.50, 15.54 and 12.00% growth inhibition in
S. commune, C. puteana, F. proliferatum, F. oxysporum and
A. alternata respectively. With subsequent increments in
concentration of Neem oil the growth inhibition rate of
each fungus also increased. The 50% of growth inhibition
was achieved in F. proliferatum at very low concentration of
0.25% only. While same extent of inhibition was recorded
for S. commune at 2.0% concentration and for C. puteana and
F. proliferatum at 4.0%. Complete inhibition for A. alternata
and C. puteana occurred at the highest concentration taken
in the experiment i.e., 10.0% while for S. commune it was
achieved at 8.0%. Complete inhibition was achieved for both
the soft rot fungi at lower concentration than the other test
fungi i.e., 6.0% (Fig. 2 and 3).

In present investigation, the inhibitory effect of Neem

oil was determined against the wood and bamboo decaying
white rot, brown rot, soft rot and sap stain fungi at various
concentrations using the food poison technique (Table 1).
Neem oil was found to be effective against all the tested fungi in
the present investigation. At the lowest tested concentration
of Neem oil i.e., 0.25% there was no inhibition in the growth Fig. 1: Growth Percent Inhibition of Bamboo Decaying
of S. commune whereas little inhibition as 5.0% and 2.5% Fungi by Neem Oil
was recorded in C. puteana and F. proliferatum respectively.

Table 1: CRD Factorial Analysis of Percent Growth Inhibition of Fungi at Different Concentrations of Neem Oil
Neem Oil Concentrations Fungal Strains (A) Mean B
(%) (B) S. commune C. puteana F. roliferatum F. oxysporum A. alternata
Positive Control (DMSO) 2.20 5.00 2.50 15.54 12.00 7.45
0.25 11.48 10.56 3.2 55.56 14.57 19.07
0.50 31.48 15.87 6.48 61.11 32.14 29.42
0.75 42.41 22.54 20.74 63.89 36.43 37.20
1.00 43.33 28.00 27.78 68.33 39.28 41.34
2.00 50.37 36.00 44.81 79.91 42.85 50.79
4.00 63.70 58.89 62.41 90.46 46.42 64.38
6.00 65.06 60.55 100 100 50.00 75.12
8.00 100 77.78 100 100 56.00 86.76
10.00 100 100 100 100 100 100
Mean A 51.003 41.519 46.792 73.48 42.969
Factors C.D @ 0.05 SE(d) SE(m)
Factor(A) 0.599 0.303 0.214
Factor(B) 0.847 0.429 0.303
Factor(A × B) 1.893 0.959 0.678

49
 Rawat, Sahoo, Hegde and Kumar
CRD two factorial analyses of the results of percent
fungal growth inhibition shows that increase in Neem oil
concentrations signi icantly reduced the growth in all ive
fungal strains. Minimum inhibitory concentrations for
both the Fusarium strains was 6.0%, 8.0% for S. commune.
Likewise for A. alternata and C. puteana was found to be
10%. However, the results also revealed that Neem oil
was effective even at lower concentrations (4.0%) for the
inhibition ofgrowth of Fusarium compared to other fungal
strains. Overall, Neem oil concentrations above 1.0%
showed a steep reduction of fungal growth. Neem oil-
induced growth inhibition was
Fig. 2: Growth inhibition of Neem oil against
Alternaria alternata
Fig. 3: Growth Inhibition of Neem Oil against
Coniophora puteana
highest in Fusarium strains followed by S. commune even
at lower concentrations whereas A. alternata and C. puteana
needed slightly higher concentrations to be effective.
DISCUSSION
The Neem oil was found to be signi icantly effective in
inhibiting the growth of bamboo decaying fungi. The results
obtained were in conformity with the indings reported
by in other studies (Al-Abed et al. 1993; Quasem et al.
1996; Amadioha 1998; Amadioha 1999; Amadioha 2003;
50
Steinhauer 1996; Al-Hazmi 2013; Kazmi et al. 1995; Ilyas
et al. 1997; Shivpuri 1997; Mondali et al. 2009; Dubey et al.
2009). The Neemseed oil completely inhibited the growth
of fungi viz., F. moniliforme, Aspergillus niger, Drechslera
rostrata and M. phaseolina at 10.00 % (Niaz et al. 2008). Niaz
and Kazmi (2005) had found that Neem oil was effective
against Aspergillus species. Singh et al. (1980) reported
fungicidal impact of Neem oil against four pathogenic fungi
while Singh et al. (1993) found the radial growth inhibition
of S. rolfsii by Neem oil.
The active ingredients of Neemare mostly of
triterpinoides, e.g., nimbin, nimbidine, azadirachtin etc.
Neem contains varied chemical constituents such as
nimolicinol, isolimolicinolide, azadirachtol, nimlinone,
nimbocinol, nimocin etc (Brahmachari 2004; Tewari 1992).
The azadirachtin component of Neem has been reported to
be as effective fungicidal as bavistin and mancozeb (Dubey
et al. 2003).
Mizoram is well known for utility of Bamboo for both
short term and long term uses. At the same time climatic
conditions are highly suitable for fungal degradation.
Therefore, it becomes invariably important to protect
the raw material and products to enhance the service life.
Neem oil being an effective and eco-friendly preservative
as supported by the indings of present study and several
other studies, there is scope for further testing with
various combinations and against broad groups of wood
decaying fungi.
ACKNOWLEDGEMENT
Authors are thankful to the HOD, Department of Forestry,
Mizoram University, Aizawl for providing the laboratory
facilities to conduct the experiments.
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