Comparative Efficacy of Some Plant Extracts against Fungal Deterioration of Stucco Ornaments in the Mihrab of Mostafa Pasha Ribate, Cairo, Egypt
The demand for medicinal plants is expanding rapidly where plants are now recognized as a safe, efficient and in expensive fungicides for treatment of stucco ornaments. In this study, three different plants extracts Anethum graveolens, Cymbopogon citrates and Juniperus oxycedrus occurring naturally in the plant essentials were evaluated to inhibit the fungal growth of the stucco ornaments in the Ribate of Mostafa Pasha (Azdomor Al Salehy) that belonged to the Ayyubid period. Three fungal species namely: Fusarium oxysporum, Aspergillus niger, Alternaria alternate were isolated from different sites on the tested stucco ornaments. The toxicity of the three plant extracts concentrations against fungal growth revealed that Cymbopogon citrates at 100% recorded the highest significant inhibition zones (6.92, 8.0 and 5.19 cm) in all tested fungal species (A. niger, F. oxysporium and A. alternate), respectively. It is the most effective one in stopping fungal deterioration followed by Anethum graveolens fungicide which only retarded fungal growth while Juniperus oxycedrus was least effective.
Received: March 27, 2011;
Accepted: August 03, 2011;
Published: January 07, 2012
Growth of microorganisms on decorative stucco cause aesthetic and structural
damage. Stucco may be colonized by communities of organisms that interact with
the substrate at different stages (Ciferri, 1999). It
must be pointed out that the biodeterioration of a given artifact commonly results
from the complex interaction established by the microorganisms co-existing simultaneously
(Warscheid and Braams, 2000). Suitable temperatures
and moisture content combined with rich nutrients in the medieval substrate
create suitable conditions for reproduction of quiescent fungal spores. Humidity
and heat permit for hyphae germination that penetrate the substrate, with mechanically
destructive effects (McNamara and Mitchell, 2005).
As colonization proceeds, the smooth surface of the painting is modified: Pigments
initially compact and resistant to attack by various agents become rough and
bioreactive (Giullitte, 1995). These events give rise
to exfoliation, cracking and loss of the coloured patina (Ciferri,
1999). Certain deteriogenic fungi change cell surface molecules, causing
genetic recombinations (Vestrepen et al., 2004).
Their metabolic products cause further chemical damage (Angell
and Chamberlain, 1991). The capacity of fungi to dissolve carbonates depends
on available carbon sources, such as oxalic and citric acids which may mobilize
cations with chelating activity (Hirsch et al., 1995).
Fungi are an important constituent of microbial endolithic assemblages in moist
ecosystems (Golubic et al., 2005). Growth and
viability of fungi on building materials under moistening and drying conditions
were investigated by Pasanen et al. (2000).
Several preventive and remedial methods have been used in tropical environments
for control and eradication of microorganisms on stucco monuments. Remedial
methods are aimed at the direct elimination by chemical treatments to eliminate
and control the growth of biodeteriogens (Curri, 1978;
Geweely and Afifi, 2011). Fungal damage on archaeological
building materials and their control measures in some museums was studied, where
Aspergillus flavus, A. versicolor, Aureobasidium sp. and
Penicillium frequentans was recorded by Cepero et
al. (1992) and Cladosporium sp. was detected by Warscheid
et al. (1992) and Kumar and Sharma (1992)
evaluates biocides for preservation of architectural building materials.
Natural plant materials are important sources of some of the widely used plant
extracts (Salehi Sourmaghi et al., 2006; Thatoi
et al., 2008; Ajibesin et al., 2008;
Keymanesh et al., 2009). Flowers, leaves, roots
and other parts are finally ground and used or the toxic principals are extracted
and utilized (Mondal and Khalequzzaman, 2010; Isman,
2006; Benamar et al., 2010). Several investigations
have been directed towards the antimicrobial activity of essential oils: (Adebajo
et al., 1989; Gangrade et al., 1991;
Kishore et al., 1980; Mishra
et al., 1990; Skukla and Tripathi, 1987).
Leaf extract of lemongrass was significantly inhibited the radial growth of
Aspergillus niger, Fusarium moniliforme (Zaman
et al., 1997; ChemMatters, 2004).
The aim of the present study was to investigate the fungal deterioration phenomena
of stucco ornaments in Ribate of Mostafa Pasha, Cairo, Egypt which suffer from
severe fungal deterioration. Hence, preservation of this material from fungal
biodeterioration by using safe fungicide was a national goal in this work.
MATERIALS AND METHODS
Evaluation of the possibility of using plant extracts especially botanical
fungicide in preservation of the tested stucco was carried out. Sensitivity
of isolated fungal species against three different plant extracts was tested.
Source of isolation and field observation: Ribate of Mostafa Pasha has
many stucco decoration in the mihrabs. These decorations suffered from different
forms of deterioration grades as discoloration black to dark green due to fungal
colonies. It is clear and scattered at different parts particularly in the damped
parts. Many cracks and lost parts at different altitudes in the stucco decorations
Isolation and identification of fungi from tested sample: Samples from
markedly damaged stucco surfaces were collected. Swabbing with sterile cotton
swabs and scalpel from markedly damaged surfaces with visible colonies of microscopic
fungi was carried out. In the laboratory, swab samples were shaken mechanically
for 10 min in 10 mL sterile distilled water and 1 mL aliquots of the resulting
suspensions used to prepare spread plates on Czapecks Dox agar in order
to isolate as wide a range of microfungi as possible. Plates were incubated
in the dark at laboratory temperature (25°C) for 7 days and the microscopic
fungi were identified using the diagnostic keys of Booth (1977),
Gilman (1957), Barnett and Hunter (1972)
and Moubasher (1993).
Plant extracts inhibition zone: Control of fungal growth on stucco decoration
from the mihrab of Mostafa Pasha Ribate by plant extracts inhibition zone was
performed. Three plant extracts were used in protecting stucco from biodeterioration
namely: Anethum graveolens, Cymbopogon citrates, Juniperus
oxycedrus. Aliquots of about 15 mL of Czapecks Dox agar were dispersed
into sterile petri dishes.
|| Surveys of genera and species of fungi isolated from stucco
Each dish was fissured by using sterilized cork borer (1 cm diameter hole)
at its center and 1 mL of each plant extract was added.
Each plant extract was used in five concentrations 10, 30, 50, 70 and 100%
and was put in each hole and each petri dish was inoculated with tested fungi
which cut from the colony margin of 2-4 day old cultures. The plates were incubated
for 24 h intervals, after which the diameters of inhibition zone (in cm) were
RESULTS AND DISCUSSION
Frequency of occurrence of fungal species isolated from Stucco Ornaments
in the Mihrab of Mostafa Pasha Ribate: Three deteriorating fungal species
were isolated from different sites on stucco. Genus Aspergillus contributed
the broadest spectra where A. niger showed highest occurrence as it occured
in the all tested sites on stucco. This result agree with Abdel-Hamid
and Ouf (1990) who recorded that Aspergillus genus was the broadest
deteriorating factor causing damage of materials and also Bisht
(1995) said that the most common bio-deteriorating fungal species were Aspergillus,
Cladosporium, Alternaria and Penicillium. On the other hand,
Fusarium oxysporum was isolated in moderate occurrence and Alternaria
alternata was low occurrence (Table 1).
It is distinctly possible that the minerals may have been contaminated by airborne
spores. Hyvarinen et al. (2004) stated that mold
growth of Aspergillus spp. son moistened damage building materials with
numbers was between 10 and 108 cfu g-1 and associated
with adverse health effects. Eight microfungi on 21 different types of building
material was investigated and only Penicillium, Aspergillus were
a dominant species (Nielsen et al., 2004). The
record of fungal occurrence on the building material points to the presence
of dung (Van Geel et al., 2003). Fungi were isolated
from the damaged building materials could use organic substances of restoration
material as nutrient sources for growth on inorganic building materials, it
is in fact the interrelationships which play the main role within microbial
communities of this type (Karpovich-Tate and Rebrikova,
Comparative assessment of microbiological deterioration of constructional material
which is part of the cultural heritage was achieved by Herrera
et al. (2004).
Plant extracts sensitivity: Three plant extracts namely: Anethum
graveolens, Cymbopogon citratus and Juniperus oxycedrus have
been applied on the isolated mould colonization from deteriorated stucco decorations.
The data revealed that a significant decrease in the growth of all isolated
fungal species with progressive increasing in the concentration of the all three
tested plant extracts. The lowest concentrations (10-30%) of the tested A.
graveolens and J. oxycedrus extracts have no effect on the growth
of three tested fungal species.
The most resistant tested fungal species was Alternaria alternata, where
the lowest inhibition zone (2.1 cm) was occurred with 100% concentration of
A. graveolens extract, on the other hand Fusarium moniliform was
considered the most sensitive tested fungal species, where the maximum significant
inhibition zone (8.0 cm) was showed with C. citratus at 100% concentration
(Fig. 1a, b).
|| Effect of different concentration of (a) Cymbopogon citrates,
(b) Juniperus oxycedrus and (c) Anethum graveolens, extracts
on the growth of the three isolated dateriorated fungal species, LSD at
0.05 = 0.50, 1.54 and 2.11, respectively
This may refer to the spores of Alternaria are pigmented and more resistant
to the inhibitory effect of the tested plant extracts where certain correlations
between morphology of the spores and their susceptibility to inhibitor were
indicated. This finding agree with that obtained by Geweely
(2009) and Hibben and Stotzky (1969) who stated
that the spores were more sensitive to inhibition are relatively hyaline, while
the most resistant spores were large and pigmented.
Fungicides sensitivity on the isolated fungal species (fungicides inhibition
zone): The obtained data revealed that Cymbopogon citrates at 100%
recorded the highest significant inhibition zones (6.92, 8.0 and 5.19 cm) in
all tested fungal species (A. niger, F. oxysporium and A. alternata,
respectively) Fig. 1a. The obtained data was agree with that
obtained by Fiori et al. (2000) who stated that
the plant species responsible for the greatest inhibition of the mycelial growth
of microorganisms was C. citratus. The highest inhibitory effect of Cymbopogon
citratus may be referred to the presence of antifungal protein in the extract.
Linthorst and Van Loon (1991) stated that low molecular
weight proteins or peptides in the plant extract have antifungal activities.
They were believed to be involved in a defense mechanism against pathogenic
fungi by inhibiting microbial growth through binding to chitin or increasing
the permeability of the microbial membranes or cell wall. According to the explanation
of Farag et al. (1989), it appears that there
is a relationship between the chemical structure of the most abundant compounds
in the plant extract and the antimicrobial activity. Fiori
et al. (2000) indicated that the plant species responsible for the
greatest inhibition of the mycelial growth of microorganisms was Cymbopogon
citratus. Tzortzakis and Economakis (2007) tested
the antifungal activity of Cymbopogon citratus oil against Aspergillus niger.
According to the explanation of Farag et al. (1989),
it appears that there is a relationship between the chemical structure of the
most abundant compounds in the essential oils and the anti-microbial activity.
The data in Fig. 1b showed that applying of 100% J. oxycedrus
on the tested fungi gave the moderate inhibitory effect. On the other hand
A. graveolens had low inhibitory effect against all tested fungal species
(Fig. 1c). Building materials are shown to be deteriorated
by a variety of fungi and the control was achieved by Nalli
et al. (2006), Schieweck et al. (2005),
Do et al. (2005) and Clausen
and Vina (2005). Tzortzakis and Economakis (2007)
tested the antifungal activity of Cymbopogon citratus oil against Aspergillus
niger. Differences in sensitivity to fungicide also existed between strains
of the same species (Dannenhauer et al., 1983).
Mostafa Pasha Ribate belonged to the Ayyubid period has many stucco ornaments
in the two mihrabs. These decorations have many colord spots due to some fungal
deterioration. Three of plant extracts were examined against the deteriorated
tested fungal species. Cymbopogon citratus was the best one for controlling
all tested fungal species. Treatment of the stucco ornaments of the mihrab must
be carried out by applying Cymbopogon citrates extract, where the highest
inhibition zones were showed in A. niger, F. oxysporium and A.
alternata, respectively. It was the most preferable inhibitor to prevent
fungal growth instead of using chemicals fungicides to avoid the unfavorable
side effect of chemicals on the properties of archeological stone and public
health and environment.
I would like to express my deepest thanks to Dr. Neveen S.I. Geweely, Associate
professor of microbiology, Department of Botany, Faculty of Science, Cairo University
her assistance and helpful discussions.
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