Intern
national Journal
J
of Phytom
medicine 7 (2015) 396-401
3
httpp://www.arjoournals.org/inndex.php/ijpm
m/index
Originaal Researchh Article
ISSN: 0975-0185
Studies on
o antimiicrobial effect of natural dyes and pigments obtained
o
from the
leaf of Art
rtocarpuss heterophhyllus Lam
m. and Tectona
Te
grrandis L.ff.
Basanti Majhhi1, Kunja Bihaari Satapathyy2*, Sagar Kum
mar Mishra3 and
a Ashirbad Mohapatra4
*Corrrespondingg author:
Kuunja Bihari Satapathy
1,2P.G
G
Department of Botany, Utkkal
University, Vani Vihaar, Bhubaneswaar7510004,
Odisha, India
3P.G. Department of Pharmacognos
sy,
University,
Utkal
Vani
Vihaar,
Bhubaaneswar-7510044,
Odisha, India
4Shri Jayadev Colle
ege of Educatioon
and
Technology,
Naharkantta,
Bhubaaneswar-7521011,
Odishha, India
Abstract
At preseent there is a reenewed interest for value additioon of textiles by using natural products. Many
of the plant
p
materials, from which nattural dyes are obtained,
o
also foound to have soome medicinal
values. However, it is a matter of conccern that the inddigenous knowleedge of extractioon, processing
and praactice of using natural
n
dyes hass diminished to a great extent among
a
the new
w generation of
ethnic people
p
due to eaasy availability of
o cheap synthetic dyes. In view of this, the pressent study was
undertaaken so as to revvive and popularrise the age-old art
a of dyeing withh natural dyes.
During the investigation dyeing materials were prepaared from leavess of Artocarpuss heterophyllus
Lam. (Jack fruit) and Tectona
Te
grandis L.f.
L (Teak). Silk fabrics
f
were dyeed with the extraacted colouring
materiaals and were esttimated for theirr wash fastness to ensure the durability
d
of the colour on the
fabrics. Finally, the antimicrobial effect of the two diffeerent natural dyeed fabrics was studied
s
against
Klebsieella pneumoniae,
e, Escherichia coli,
co Candida albbicans and Aspeergillus sp. The dye prepared
from leaaves of Tectonaa grandis L.f. waas found to be most effective by
b showing maxximum zone of
inhibitioon as compared to Artocarpus heterophyllus
h
Laam. The textile material
m
impregnnated with two
natural dyes resulted in the maximum
m zone of inhibbition rate (65%
%) against Esche
herichia coli as
recordeed in the silk sam
mples dyed with Tectona
T
grandiss.
It can be
b concluded thaat the natural dyyes obtained from
m these two plaants under studyy could provide
special aesthetic valuess, which is not only
o environmenntally friendly buut gives added qualities
q
to the
textile production
p
becauuse of the antimiicrobial potentiall of the dyes.
Keyworrds: Cisplatin Anntimicrobial activity; Artocarpuss hetphyllus Lam
m.; Tectona graandis L.f.; Silk
fabric; Mordants;
M
Disc diffusion.
d
Inntroductionn
Naatural dyes haas gained impportance due to the growinng
ennvironmental aw
wareness and due to the reesult of stringeent
ennvironmental staandards imposedd by many counntries in responsse
to toxic and overaall water quality issues due to efffluents associateed
with synthetic dyes. Under such situation
s
natural dyes
d
derived froom
plaants have gainned economic advantage oveer synthetic dyees
beecause of their non-toxic, non--carcinogenic and biodegradabble
naature.[1] Up to thhe end of 19th ceentury natural dyyes were the maain
coolourants for texttiles. Recently, interest in the usse of natural dyees
haas been growing rapidly duue to the ressult of stringeent
ennvironmental staandards imposedd by many counntries in responsse
to toxic and allerrgic reactions associated with synthetic dyes.[[2]
Naatural dyes maay have a widee range of shaades and can be
b
obbtained from varrious parts of plaants including rooots, bark, leavees,
floowers, and fruit. [3]
Prresently there is an excessive usse of synthetic dyes,
d
estimated at
aroound 10,000,000 tons per annum, the productioon and applicatioon
of which release vast amount of waste and unfixed colorannts
caausing serious health
h
hazards and
a disturbing thhe eco-balance of
natuure. [4] Now-a-ddays, fortunatelyy, there is increaasing awarenesss
amoong people tow
wards natural dyes.
d
Natural dyyes have betteer
bioddegradability annd generally havve higher comppatibility with thee
envvironment. Theyy are non-toxiic, non-allergic to skin, nonncarccinogenic, easilyy available and renewable.
r
[5]
Nattural dyes are peermanent as coompared to synthhetic colorant. Inn
Indiia, Rajasthan, Orissa
O
(Kotapadd, Nuapatana, and
a Berhampurr)
andd Kutch still posssess a rich tradittion in the use off natural dyes foor
texttile dyeing. The widely and commercially usedd synthetic dyess
imppart strong colouurs but causes carcinogenicity and inhibition oof
bennthic photosynthesis. [6] Thus, Inndia harbours a wealth of usefuul
germplasm resourcces and there is no doubt that thhe plant kingdom
m
is a treasure-housee of diverse natuural products. One
O such producct
from
m nature is the dye.
d Mordants are
a metal salts which
w
produce ann
affinnity between thee fabric and the dye.
d [7]
Currrently, both natural
n
and syynthetic fibres have inherennt
resiistance to miccroorganisms. Textile materiaals provide ann
exccellent environmeent for microorgaanisms to grow, because of theiir
largge surface area and ability to reetain moisture. Microbial activityy
cann be detrimental to textiles. It caan cause unpleaasant odour, leadd
to weakening
w
of thee substrate, discoloration, and evven contribute too
the spread of diseaase. For this reeason, antimicroobials have beenn
Th
his work is lice
ensed under a Creative Com
mmons Attribution 3.0 License.
Majhi et al. Innternational Joournal of Phyttomedicine 7 (4) 396-401 [2015]
invvestigated as a finish for textilles. [8] Antibactterial finishes are
a
appplied to textiles for three major reasons: (a) to curtail the spreaad
of disease and avvoid the danger of injury-induceed infection, (b) to
cuurtail the developpment of odour from
f
aspiration, stains and soil on
o
texxtile materials, and (c) to currtail the deteriooration of textilees
caaused by mildew
w, particularly fabbrics made of naatural fibres. [9] In
ordder to obtain thee greatest benefit, an ideal antim
microbial treatmeent
of textile should saatisfy a number of requirements; [10] of these, thhe
anntimicrobial agennt should be effective against broad
b
spectrum of
baacterial and fungal species butt at the same time exhibits loow
toxxicity to consum
mers, e.g., not caause toxicity, alleergy or irritation to
thee user. Naturall dyes can produce special aesthetic qualitiees,
whhich, combined with the ethical significance off the product thhat
caan evolve into ann eco-friendly and value added teextile fabrics.
Thhe present studyy was undertakeen with an objecctive to revive thhe
agge-old art of natural
n
dyeing apart from thhe study of thhe
anntimicrobial potenntial present in the natural dyes obtained from thhe
leaaves of Artocarppus heterophyllus
us and Tectona grandis
g
.
M
Materials
and Methodds
Figure 1 A: Artoocarpus heterophhyllus Lam.
Collection of plaant materials for the experim
ment
Arto
tocarpus heteropphyllus Lam. (JJack fruit) leavees and Tectona
na
graandis L.f. (Teaak) leaves colleected from Balaasore district oof
Odisha (India); 6.5 cm silk fabrics were
w used for dyyeing.
Arto
tocarpus heteropphyllus Lam. (Mooraceae) is a laarge tropical treee
andd is indigenous to India and popularly know
wn as Jack treee
(Figgure 1 A, B). The
T leaves aree simple, alternate, coriaceouss,
entiire, dark, shiny green
g
above, obblong, oval or eliiptic in form, 4 too
6 innches in length, glabrous, hairless with smooth skinned surfacee.
Thee leaves show the presence of sapogenin, cycloaratenonee,
cycloartenol, sitosteerols and tanninns. [11] The leavves are reportedd
to be
b used in skin diseases and assh of leaves is useful in healingg
ulceer. The leaves are also usefuul in boils, wouund, fever, skinn
diseeases and vitiateed condition of pitta
p and vata. [12]
Tecctona grandis L..f., commonly known
k
as teak, is a widespreadd
wooody plant with loot of biological properties.
p
[13] Large
L
deciduouss
treee, branchlets 4-aangled. Leaves large, opposite, simple, obovateeellipptic, stellately (F
Figure 2 A, B); useful
u
in skin diseases, leprosyy,
coooling, diabetes, and
a bronchitis. [114]
hyllus Lam.
Figure 1 B: Colour substannces extracted frrom A. heterophy
PAGE | 3977 |
Majhi et al. Innternational Joournal of Phyttomedicine 7 (4) 396-401 [2015]
Figure 2 A: Tect
ctona grandis L.f..
Exxtraction of dyye
Exxperiments weree conducted for extraction of thee natural dye wiith
ann effort to get the maximum amount from the
t leaves of A.
heeterophyllus andd T. grandis. Samples
S
collecteed were dried at
rooom temperaturee (25%) compleetely before groound in the mixxer
grinder. 500 g of the
t leaf powder was taken in a soxhlet apparatuus
treeated with 35000 ml of 10 % aqqueous ethanol. On heating, thhe
whhole extracts unnderwent the proocess of distillaation to obtain thhe
cruude dye as a paasty mass. The pasty extract was
w washed with a
noon-polar solventt such as petrooleum ether followed by freezze
dryying to obtain the colouring maaterials in solveed form. The finnal
yieeld of material in case of Artoccarpus heterophy
hyllus and Tecton
ona
gra
randis was fouund to be 70 g (14%) and 74 g (14.80%
%)
resspectively, whichh was used as thhe experimental material.
Prreparation of standard dye solution
Thhe semi-solid dye materials were converteed into differeent
cooncentration of standard solution by 10% aqqueous methannol
soolvent. Thus staandard dye soluutions were preppared at differeent
cooncentrations such
s
as 10%
%, 8%, 6%, 4%, and 1%
%.
Dyyeing procedure Dyeing was donne with or withouut mordant as well
ass pre-mordantingg and post-mordanting.
Arrtocarpus heteerophyllus
Laam.For the purppose the fabric silk was dippedd in the prepareed
moordant solution 20 ml (10 % Alum + turmeric powder)
p
for 30-445
miinutes at 50-60 0c. Then the moordanted silk waas air dried for 15
1
Figure 2 B: Colour
C
substance extracted from
m T. grandis L.f.
minnutes. The dye solution
s
(20 ml) was
w prepared with
w 0.2% sodium
m
hyddroxide solution. The mordantedd fabric was theen dipped in thee
dyee bath for 30-45 minutes at 60-700 0c for silk. Thee dyed fabric wass
left for 15 minute for
f air oxidation. The golden yeellow colour wass
achhieved.
Tecctona grandiss L.f.
Thee silk fabric wass dipped in the prepared
p
mordant solution of 200
ml (5%
( CuSo4) for 30-45 minute at
a 50-60 0c. Thenn the mordantedd
silk was air dried for 15 minute. The dye solutiion (20 ml) wass
prepared with 0.2%
% sodium hydrroxide solution. The mordantedd
fabrric was then dippped in the dye bath for 30-45 minutes
m
at 60-700
0c for
f silk. The dyeed fabric was lefft for 15 minute for air oxidationn.
Thee brick red colour was achieved.
Tesst organism
Cultures of followiing microorganiisms were useed in the studyy:
Kleb
ebsiella pneumonniae ATCC 17055, Escherechia coli
c ATCC 10533,
Canndida albicans ATCC 10231, Aspergillus spp. MTCC- 87900
obtaained from Himeedia were used in this study.
Determination of
o antimicrobiaal activity of natural
n
dyes inn
solution
Discc diffusion meethod was usedd to screen thhe antimicrobiaal
activities. The nutrieent agar plates were prepared by
b pouring 15 m
ml
of media
m
into sterilee Petri-plates annd in the same way PDA platess
werre also preparedd. The plates were
w
allowed to solidify for 2 hrss
andd then preparedd broth of 0.1% inoculums suspension
s
wass
streeaked uniformly. Sterile filter paper disc (diametter 5.0 mm) wass
PAGE | 3988 |
Majhi et al. International Journal of Phytomedicine 7 (4) 396-401 [2015]
placed on top of the seeded medium. 30 μl of different
concentration of Tectona grandis and Artocarpus heterophyllus
(1%, 4%, 6%, 8%, 10%) dye solution were loaded to diffuse for 2
minute and then the plates were kept for incubation at 37 0c for
overnight (16 hrs) for bacteria. For fungi all procedure was similar
but 50 μl dye solution was loaded on the filter disc and it was
allowed to diffuse for 2 minute and the plates were kept for
incubation at room temperature for 16 hrs. Inhibition zones formed
around the disc were measured with ruler in centimetres. All the
studies were performed in triplicates.
The results are reported in the Table 1 and Table 2. The data on
the efficacy of the natural dyes upon the test organisms revealed
that at 10%, 8%, 6% and 4% concentrations of the dye obtained
from Artocarpus heterophyllus resulted in the significant inhibition
zone. The results also indicated that there was a gradual increase
in the zone of inhibition with the increase in the concentration of the
leaf extract from 4% to 10% irrespective of the test organisms.
However, at 1% concentrations, mild growth of microbes was
observed along with zone of inhibition. Artocarpus heterophyllus
was more effective against Escherichia coli as compared to other
test organisms. However, Artocarpus heterophyllus was found to
be less effective against Candida albicans.
Screening test for Tectona grandis showed that the dye was
effective against all the four microbes irrespective of the
concentration of the extract. Highest zone of inhibition could be
seen against Klebsiella pneumoniae as a typical example as
mentioned in Table 2. The results also indicated that there was a
gradual increase in the zone of inhibition with the increase in the
concentration of the leaf extract from 4% to 10% irrespective of the
test organisms. However, a mild growth at 1% was observed
thereby indicating that the dye was not effective to control the
growth of the microbes at lower concentration.
The antimicrobial potential of both the dyes were studied by
observing the zone of inhibition as presented in Table 1 and Table
2, which indicated that the increase in the concentration of dye
could lead to increased inhibition as reflected by enhancement in
diameter. It can be concluded that both the dyes could be used as
effective antimicrobial agents. It is evident from Table 1 and 2 that
with increasing concentrations of the dye, the zone of inhibition is
increased almost linearly. The increase in zone width is relatively
larger for Artocarpus heterophyllus than Tectona grandis, thus it is
apparent that the dyes evaluated during the present study have
bactericidal and fungicidal properties.
Determination of antimicrobial activity of dye fabrics
In this experiment the antimicrobial activity of dyed fabrics was
tested. The 6.5cm fabric (dyed and undyed for control) was
introduced in the 100 ml nutrient broth inoculated with the desired
test pathogens and incubated at 37 0c over night for 24 hrs for
bacteria and fungi for a room temperature for 16 hrs. The reduction
in the growth of bacteria and fungi by the dye was expressed as
follows.
R = B - A/B x 100
Where:
R = % Reduction in bacterial population
B = Absorbance (660 nm) of the media inoculated with microbe
and undyed fabric.
A = Absorbance (660 nm) of the media inoculated with microbe
and dyed fabrics.
Results and discussion
Screening of natural dyes for anti-microbial activity
Solution of different concentrations of the two natural dyes under
study was screened for their activities against the test organisms.
Table 1: Antimicrobial activity of natural dye (inhibition zone in cm) from leaves of Artocarpus heterophyllus Lam.
Dye
Sl.N
Test organisms
Measurement of inhibition zone (in cm)
o
Different conc. of plant extracts
Artocarpus
heterophyllus
1%
4%
6%
8%
10%
1
Klebsiella pneumoniae
0.71μ0.01
1.43μ0.02
3.0μ0.08
3.92μ0.11
4.56μ0.12
2
Esherichia coli
0.69μ0.00
1.2μ0.01
2.8μ0.04
3.57μ0.08
4.6μ0.14
3
Candida albicans
0.56μ0.01
0.97μ0.01
1.56μ0.02
2.33μ0.03
2.53μ0.03
4
Aspergillus sp.
0.73μ0.01
1.47μ0.01
3.3μ0.28
4.36μ0.12
5.16μ0.28
PAGE | 399 |
Majhi et al. Innternational Joournal of Phyttomedicine 7 (4) 396-401 [2015]
Table 2: Antimicrobiaal activity of natural dye (inhibitioon zone in cm) frrom leaves of Teectona grandis L.f.
L
Dyye
Sl.N
No
Test organisms
Meeasurement of innhibition (in cm)
Different conc. of plant extractss
Tectonaa grandis
1%
1
Klebsiellaa pneumoniae
0.7μ0.01
2
Esherichi
hia coli
0.69μ0.00
Candida albicans
a
3
4
Aspergillu
lus sp.
0.56μ0.01
0.73μ0.01
Anntimicrobial activities
a
of Fabrics
F
treateed with naturral
dyyes
Since dyes showeed good antimiccrobial activitiess against selecteed
miicrobes in solution, it was thought worthwhiile to study theeir
anntimicrobial activvities on dyed suubstrate (fabric). The silk samplees
dyyed with these natural
n
dyes were used as moddel system. It waas
revvealed from Figure 3 that the dyye obtained from
m Tectona granddis
4%
6%
8%
10%
1.36μ0.04
2.76μ0.12
3.333μ0.32
4.336μ0.33
1.16μ0.04
1.93μ0.20
2.76μ0.24
4.006μ0.16
1.2μ0.08
2.3μ0.21
3.443μ0.41
3.886μ0.12
2.56μ0.09
3.22μ0.24
4.00μ0.21
1.23μ0.09
wass 50% more effficient than the dye obtained from Artocarpus
us
hete
terophylus with regard
r
to the coontrol of Klebsieella pneumoniaee,
Esccherichia coli andd Candida albica
cans, where as foor Aspergillus spp.
bothh the dyes weree found to be more
m
or less equually effective. IIt
wass evident from Table 3 that the
t dye from Tectona
T
grandis
is
reduced the growthh in case of Eschherichia coli up to
t 65% where ass
the dye obtained from Artocarppus heterophyllu
lus reduced thee
grow
wth of Aspergilluus sp. up to (44%
%).
70
60
50
40
30
Klebsiella pneumoniae
20
Escherichia coli
10
Aspergilluss sp.
0
Candida allbicans
randis
Figuree 3: Antimicrobiaal activity of textile materials dyed with Artocarpuus heterophylus and Tectona gra
PAGE | 4000 |
Majhi et al. International Journal of Phytomedicine 7 (4) 396-401 [2015]
Table 3: Antimicrobial activity (reduction of growth in %) of the fabrics treated with natural dyes of the plants.
Test organism
Initial Absorbance (O.D.)
Klebsiella pneumoniae
Escherichia coli
Candida albicans
Aspergillus sp.
0.486
0.736
0.300
0.282
Conclusion
The dyes obtained from the two plants (Artocarpus heterophyllus
Lam. and Tectona grandis L.f.) used in the current study were
found to have effective antimicrobial potential against the four
microbial pathogens i.e. Klebsiella pneumonia, Escherichia coli,
Candida albicans and Aspergillus sp. The dye obtained from
Tectona grandis was 50% more effective to control the growth of
Klebsiella pneumoniae, Escherichia coli and Candida albicans as
compared to the dye obtained from Artocarpous heterophyllus. The
Growth of test microbes
Absorbance with dyed fabrics
(O.D.)
Reduction in growth of test
organisms (%)
Artocarpus
heterophylus
Tectona
grandis
Artocarpus
heterophylus
Tectona
grandis
0.328
0.550
0.256
0.157
0.204
0.258
0.187
0.176
32.510
25.271
14.66
44.33
58.024
64.945
37.66
37.58
efficacy of both the dyes in respect of the control of Aspergillus sp.
was observed to be nearly similar. The effectiveness of the both
the dyes were observed to increase linearly with the increase in
concentration of the dye from 4% to 10%. It can be concluded that
the natural dyes obtained from these two plants under study could
provide special aesthetic values, which is not only environmentally
friendly but gives added qualities to the textile production because
of the antimicrobial potential of the dyes.
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