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Mycosynthesis of Silver Nanoparticles Using the Fungus Fusarium acuminatum and

its Activity Against Some Human Pathogenic Bacteria

Article  in  Current Nanoscience · May 2008

DOI: 10.2174/157341308784340804

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 Current Nanoscience, 2008, 4, 141-144 141

Mycosynthesis of Silver Nanoparticles Using the Fungus Fusarium acuminatum and

its Activity Against Some Human Pathogenic Bacteria
Avinash Ingle1, Aniket Gade1, Sebastien Pierrat2, Carsten Sönnichsen2 and Mahendra Rai1,*
1
Departement of Biotechnology, SGB Amravati University, Amravati-444 602, Maharashtra, India; 2Johannes Gutenberg University
of Mainz, Jakob Welder Weg 11, 55128 Mainz, Germany

Abstract: We report extracellular mycosynthesis of silver nanoparticles by Fusarium acuminatum Ell. and Ev. (USM-3793) isolated
from infected ginger (Zingiber officinale). An aqueous silver nitrate solution was reduced to metallic silver when exposed to F. acumi-
natum cell extract leading to the appearance of a brown color within 15-20 minutes. The color is due to the formation of silver nanoparti-
cles and the excitation of surface plasmons. The optical spectrum showed the plasmon resonance at 420 nm and analysis by transmission
electron microscopy confirmed the presence of silver nanoparticles. The nanoparticles produced were spherical with a broad size distribu-
tion in the range of 5-40 nm with average diameter of 13 nm. The reduction of the silver ions occurs probably by a nitrate-dependent re-
ductase enzyme, which we found to be present in the extra-cellular medium. We tested the silver particles for their broad-band antibacte-
rial activity on different human pathogens. We observed efficient antibacterial activity against multidrug resistant and highly pathogenic
bacteria, including multidrug resistant Staphylococcus aureus, Salmonella typhi, Staphylococcus epidermidis, and Escherichia coli. The
synthesis of silver nanoparticles by the fungus F. acuminatum may therefore serve as a simple, cheap, eco-friendly, reliable and safe

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method to produce an antimicrobial material.

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Keywords: Fusarium, Extracellular, Mycosynthesis, Nanoparticle, Antibacterial.

INTRODUCTION We have selected Fusarium acuminatum for synthesis of silver

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New multi-drug resistant strains of bacteria have become a se- nanoparticles because it is easy to isolate from ginger, needs simple

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rious problem in public health [1, 2]. The emerging resistances in environmental conditions for growth, and can be grown on simple
bacteria and high cost of advanced antimicrobial drugs have en- potato sucrose agar medium. Fusarium acuminatum is an important
and well-studied plant pathogen [37], but its exploitation for bene-

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couraged researchers to search for effective and economically vi-
able broadly applicable drugs [3]. It has been known for long time ficial biomedical applications like the production of an antibacterial

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agent is a novel contribution.

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that silver ions are highly toxic to a wide range of bacteria, and
silver-based compounds have been used extensively in bactericidal

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applications [4, 5]. Silver has an advantage of having broad antimi- EXPERIMENTAL
crobial activities against Gram-negative and Gram-positive bacte- The fungus F. acuminatum was isolated from infected ginger

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ria. Silver nanoparticles are the most effective preparation of silver (Zinziber officinale) (Fig. (1A-D)), characterized and identified on

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because of the high surface/volume fraction resulting in a large the basis of morphological characteristics [38] which was inde-
proportion of silver atoms in direct contact with their environment. pendently confirmed by Professor Baharuddin Salleh, Department

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Besides their use in the field of antimicrobial agents, silver of Biological Sciences, University of Sains, Malaysia. The isolated

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nanoparticles find applications in biolabelling, biosensors, and fungus culture was maintained at 28±2°C on potato sucrose agar
filters [6]. Chemical, physical, and biological methods are used for slants. To prepare biomass for mycosynthesis studies, the fungal

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the synthesis of metal nanoparticles [7, 8], with chemical methods culture was grown aerobically at 28±2°C in potato sucrose broth
most widely applied because large quantities of nanoparticles are containing potato infusion (250 g/l), sucrose (20g/l). The flasks

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produced in short periods of time. However, physical and chemical
techniques tend to be capital intensive, inefficient in materials, and
energy use and may require toxic or environmentally damaging
chemicals. Nanoparticle synthesis by biological or biomimetic ap-
proaches might offer inexpensive, clean, non-toxic, and eco-
friendly alternatives. Biosynthesis of metal nanoparticles has been
demonstrated in different plants, such as Alfalfa [9, 10], Geranium
leaves [11], Aloe vera [12], Cinnamomum camphora [13]. This
‘green synthesis’ of metal particles depends on living systems such
as plants and microbes and a survey of literature indicates that
green synthesis of nanoparticles is gaining importance [14]. Mi-
crobes including algae, bacteria and fungi, are known to produce
nanoparticles either intracellularly [15-23] or extracellularly [24-
33]. Recently, Fusarium oxysporum [24-26], Pseudomonas
stutzeri [15], Rhodococcus sp. [34], Thermomonospora and
[24-26], Phaenero chaete chrysosporium [35] have been stud-
ied. Many successful attempts have been made for the synthesis of
nanoparticles of different shapes [15] and different metals, includ-
ing silver [29], platinum [30], gold [31], zirconia [32], silica and
titania [33], CdS [36].
*Address correspondence to this author at the Department of Biotechnology, SGB
Amravati University, Amravati-444602, Maharashtra, India; Tel: 91-721-2667380,
Fax: 91-721-2660949; Cell - 9422857196; E-mail: mkrai123@rediffmail.com
were inoculated, incubated on orbital shaker at 28±20 C, and then
agitated at 120 rpm. We harvest the biomass after 72 h of growth by
sieving through a plastic sieve and filter paper, followed by two to
three times washing with distilled water to remove any medium
component from the biomass. Approximately 20 g (wet weight) of
biomass was mixed with 100 ml of double distilled water for 48-72
h at 28±2 °C in an Erlenmeyer flask and agitated in the same condi-
tions as described earlier. After the incubation, the cell filtrate was
obtained by passing it through Whatman filter paper no. 1. The
resultant filtrate was treated with aqueous AgNO3 in such way that
the final concentration of the solution was 1mM. Experiment was
set up in triplicate. After incubation of 2 hours, the sample were
removed from the reaction solution and subjected to UV–Vis spec-
troscopy measurements on a Perkin-Elmer (lambda-25) spectropho-
tometer at a resolution of 1 nm from 280 to 800 nm. For the subse-
quent tests, we separate and concentrate the silver nanoparticles by
repeated centrifugation at 10000g replacing the supernatant with
distilled water. The presence of un-reacted silver ions leads to the
formation of a white precipitate upon addition of sodium chloride.
No white precipitate was formed after addition of sodium chloride
indicating the absence of unreacted silver ions in the nanoparticle
solution.
The silver nanoparticles were characterized by Transmission
Electron Microscopy (TEM). Conventional carbon coated copper
TEM grids (400 meshes, Plano Gmbh, Germany) were cleaned

1573-4137/08 $55.00+.00 © 2008 Bentham Science Publishers Ltd.

 142 Current Nanoscience, 2008, Vol. 4, No. 2 Ingle et al.

A B C

D F

1 2

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Fig. (1). Fusarium acuminatum isolated from infected ginger A. Dorsal side B. Reverse side C. Macroconidia of F. acuminatum D. Infected ginger E. Fungal

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filtrate: control (left) and treated with 1mM silver nitrate (right) F. Specific substrate disc for nitrate reductase: 1. Fungal filtrate + Substrate disc 2. Distilled
water + disc.

u
A B

rib
0.6
Extinction (a.u.)

15
1.5

t
Extinction (a.u.)

s
0.4

i
1.0

D
0.2

r
0.5

o
E

0.0

F
0.0
400 500 600 700 800 900
300 400 500 600 700 800

t
Wavelength (nm) Wavelength (nm)

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Fig. (2). A UV-Visible optical extinction spectrums of fungal filtrate containing silver nanoparticles: Blue – cell filtrate + silver nitrate, Red – biomass +
silver nitrate, Pink – filtrate B spectrum of silver nanoparticles after centrifugation.

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using plasma treatment under oxygen atmosphere for 45s. A 5
l
drop of the sample was then deposited on the grid and dried at room
temperature for 1h. The samples were inspected in a TEM (Phillips,
CM12) operating at 120KV. At least 3 images of each sample were
taken to have a clear representation of its composition.
EVALUATION OF ANTIBACTERIAL ACTIVITY
Test Bacteria
duced in the agar and 20
l of each cleaned filtrate of nanoparticles
poured into the wells. Untreated fungal filtrate was used as control.
The plates were incubated at 37 °C overnight. The experiment was
carried out in triplicate. The antibacterial activity was evaluated by
measuring the size of the clear zone around each well and compared
to the antibacterial activity of a pure silver nitrate solution.
RESULTS AND DISCUSSION

Escherichia coli (JM-103, ATCC-39403), Salmonella typhi
(MTCC-733), Staphylococcus epidermidis (ATCC-12228) and S.
aureus (ATCC-25923) were selected to assess the activity of silver
nanoparticles against bacteria because these organisms are fre-
quently involved in hospital acquired infections [39].
Assay for Antibacterial Activity
The cultures of the bacteria were maintained in nutrient agar at
370C. The antibacterial activity of nanoparticles was evaluated by
the well diffusion method [40]. We prepare the nutrient agar by
dissolving 5g peptone, 5g NaCl, 3g beef extract and 20g of agar
powder in 1 liter distilled water and pour the cooled medium into
sterile petri plates to a uniform depth of 4 mm. Once the medium
had solidified, the culture (inoculum contains 105 CFU) was inocu-
lated on the medium by treating the surface with an L-spreader (Hi-
media, Mumbai). With a sterilized 5mm cork borer, wells are intro-
After mixing the F. acuminatum filtrate with an aqueous
AgNO3 solution, we observed a color change from light yellow to
brown. The color change started about 15-20 min after mixing and
continued to intensify up to approximately 2 hours of incubation
(Fig. (1E)). The color-change indicates the formation of silver
nanoparticles, which show a yellowish-brown color in water due to
the excitation of surface plasmons in the metal nanoparticles [41].
The formation of silver nanoparticles was confirmed both by optical
spectroscopy and direct electron microscopic visualization. Extinc-
tion spectroscopy of ultraviolet (UV) and visible (Vis) light (UV–
Vis spectrum) allows to confirm the presence of noble metal nano-
particles due to the characteristic plasmon resonance. UV–Vis
spectra recorded after 2 hours of incubation are shown in “Fig. (2)”.
They show the typical plasmon resonance at 420 nm expected for
spherical silver nanoparticles [25, 26].
In order to verify the results of the UV–Vis spectral analysis,
some samples were inspected with a transmission electron micro-
 Mycosynthesis of Silver Nanoparticles Using the Fungus Current Nanoscience, 2008, Vol. 4, No. 2 143

though it was thought to be specific for F. oxysporum. The reported

mechanism, shown in “Fig. (3)”, suggests a reduction of Ag+ to Ag0
due to a conjugation between the electron shuttle with the NADH
dependent reductase [25, 26]. The actual reduction mechanism may
be due to the transfer of an electron from NADH and NADH-
dependent enzyme acting as electron carrier [23, 24]. We find ni-
trate reductase in the fungal cell filtrate using a specific substrate
disc for nitrate reductase (Nitrate Reagent Discs DD 041, Hi-media,
Mumbai, India). The color of the disc turned reddish from the white
when treated with the fungal cell filtrate (Fig. (1F)) indicating the
presence of nitrate reductase. The present study of silver nanoparti-
cle formation utilizing F. acuminatum is to our knowledge the first
such report.
Fig. (3). Possible mechanism of enzymatic reduction of the silver ions.

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rib u
is t
Fig. (4). TEM bright field image of the silver nanoparticles. (Scalebar 100 nm).

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scope. The direct electron microscopic visualization allows measur- Silver and silver-ions are known to possess antibacterial activity

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ing the size and shape of the silver nanoparticles formed. Typical [42-45]. Sondi and Salopek-Sondi [46] reported antimicrobial activ-
bright-field TEM images of the synthesized silver nanoparticles are ity of silver nanoparticles against E. coli and recommended them

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shown in “Fig. (4)”. The silver nanoparticles were more or less for the formulation of new bactericidal agents. Kim et al. [47] in-

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spherical in shape and showed a large distribution of sizes in the vestigated antimicrobial activity of silver nanoparticles against E.
range of about 5-40 nm. A particle size distribution histogram de- coli and Staphylococcus aureus. We study the efficiency of the

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termined from the TEM images shows the large variation in the silver nanoparticles produced as described above against four hu-

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particle size with about 30% of the particles in 0-5 nm range, and man pathogenic bacteria: E. coli, Salmonella typhi, Staphylococcus
7% in 40-45 nm ranges (Fig. (5)). epidermidis and S. aureus (Fig. (6)). The silver nanoparticles

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proved to be toxic to each of the above species and the effect is 1.4–
60
1.9x stronger than that of pure silver ions. Interestingly, the maxi-
mum antibacterial activity of silver nanoparticles was shown
50 against S. aureus (17mm), followed by S. epidermidis, Salmonella
typhi and the minimum by E. coli (10mm). Our findings corrobo-
40 rate the report of Shahverdi and his co-workers [48], who found
that S. aureus showed the maximum sensitivity to silver nanoparti-
Count

30 cles, whereas E. coli was less sensitive, but the contrary was re-
ported in [47]. The reason for the species-specific efficiency of
C

silver nanoparticles and possible differences from strain to strain

20 are presently unknown and the mechanism of the antibacterial ac-
tion is a matter for further investigation. It is known that the struc-
10 ture of the membranes of Gram-positive and Gram-negative bacte-
ria is different and that nanoparticles accumulate in or near the
0 membrane with some particles penetrating the membrane. The af-
5 10 15 20 25 30 35 40 finity to those membranes and the ability of releasing silver ions
into the intracellular medium may be factors influencing the degree
Particle diameter (nm) of toxicity. The molecular mechanism may be a reaction of silver
Fig. (5). Histogram of the nanoparticle size distribution extracted from with –SH groups of proteins in the cell inactivating the proteins
TEM images (183 particles counted). [49], but it remains to clarify whether and how the silver nanoparti-
cles upset the function of membranes. The effect of silver particles
In order to clarify the mechanism of silver reduction, we test for on important cellular mechanism like the synthesis of DNA, RNA
the release of a reductase enzyme into the solution by F. acumi- and proteins need to be addressed.
natum. The better-studied fungus Fusarium oxysporum is known
for the synthesis of metal nanoparticles [25-29] and a NADH de-
pendent reductase enzyme has been associated with silver reduction CONCLUSION
[25, 26]. We believe this enzyme is also responsible for the reduc- We suggest F. acuminatum as an alternative myco-biosystem
tion of Ag+ to Ag0 in the case of fungus F. acuminatum, even for the synthesis of silver nanoparticles, which may be optimized
 144 Current Nanoscience, 2008, Vol. 4, No. 2 Ingle et al.

1 2 Cell filtrate 1mM AgNO3 Filtrate + AgNO3

a a

adius of the Inhiibition

b 15
b
c c

zone (mm)
10

3 a 4 b c 5
b
0
c a E coli SS.
E. Styphi
typhi SS
S.epider
epider.SS
epider S.aureus
aureus

Ra
Tested bacteria
A B
Fig. (6). A. Antibacterial activity of silver nanoparticles against 1. Escherichia coli 2. Salmonella typhi 3. Staphylococcus epidermidis 4. S. aureus, In each
image: a. Cell filtrate, b. AgNO 3 (1mM) and c. silver nanoparticles B. Graphical representation of the size of the zone of inhibition for the four tested bacteria

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cultures.

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for better nanoparticle size and shape distribution. We demonstrated [22] Singaravelu, G.; Arockiamary, J. S.; Kumar, V. G.; Govindaraju, K. Colloids
high antibacterial activity of the silver nanoparticles produced by F. Surf. B Biointerfaces, 2007, 57, 97.
[23] Shiying, H.; Zhirui, G.; Zhang, Y.; Zhang, S.; Wang, J.; Gu, N. Mater. Lett.,

u
acuminatum that may suggest their future use in potent broadband 2007, 61, 3984.
antibacterial agents/drugs. [24] Ahmad, A.; Mukherjee, P.; Mandal, D.; Senapati, S.; Khan, M. I.; Kumar,

rib
R.; Sastry, M. J. Am. Chem. Soc., 2002, 124, 12108.
[25] Ahmad, A.; Mukherjee, P.; Mandal, D.; Senapati, S.; Khan, M. I.; Kumar,
ACKNOWLEDGEMENTS R.; Sastry, M. Colloids Surf. B Biointerfaces, 2003a, 28, 313.

t
[26] Ahmad, A.; Senapati, S.; Khan, M. I.; Kumar, R.; Sastry, M. Langmuir,
We thank Prof. Baharuddin Salleh, University Sains Malaysia,

s
2003b, 19, 3550.

i
Malaysia for confirming the identification of F. acuminatum. S.P. [27] Anilkumar, S.; Abyaneh, M. K.; Gosavi, S. W.; Kulkarni, S. K.; Pasricha, R.;
acknowledges financial support by the German science foundation Ahmad, A.; Khan, M. I., Biotechnol. Lett., 2007, 29, 439.

D
(DFG) for a visiting grand [50]. [28] Bhainsa, K.C.; D’Souza, S.F. Colloids Surf. B Biointerfaces, 2007, 47, 160.
[29] Duran, N.; Marcato, P.D.; Alves, O.L.; De Souza, G.; Esposito, E. J. Nano-

r
biotechnol., 2005, 3, 8.
REFERENCES [30] Riddin, T.L.; Gericke, M.; Whiteley, C.G. Nanotechnology, 2006, 17, 3482.

o
[31] Mukherjee, P.; Senapati, S.; Mandal, D.; Ahmad, A.; Khan, M. I.; Kumar,
[1] Wright, G.D. Chem. Biol., 2000, 7, R127. R.; Sastry, M. Chembiochem, 2002, 3, 461.

F
[2] Wright, G.D. Adv. Drug. Deliv. Rev., 2005, 57, 1451. [32] Bansal, V.; Rautaray, D.; Ahmad, A.; Sastry, M. J. Mater. Chem., 2004, 14,
[3] Jones, S. A.; Bowler, P. G.; Walker, M.; Parsons, D. Wound Repair Regen., 3303.

t
2004, 12, 288. [33] Bansal, V.; Rautaray, D.; Bharde, A.; Ahire, K.; Sanyal, A.; Ahmad, A.;
[4] Cho, K.H.; Park, J.E.; Osaka, T.; Park, S.G. Electro-chimica Acta, 2005, 51, Sastry, M. J. Mater. Chem., 2005, 15, 2583.

o
956. [34] Ahmad, A.; Senapati, S.; Khan, M. I.; Ramani, R. R.; Srinivas, F. v.; Sastry,
[5] Ip, M.; Lui, S.L.; Poon, V.K.M.; Lung, I.; Burd, A. J. Méd. Micróbiol., 2006, M. F. Nanotechnology, 2003, 14, 824.

N
55, 59. [35] Vigneshwaram, N.; Kathe, A. A.; Varadarajan, P. V.; Nachane, R. P.; Bala
[6] Cao, G (Ed). Nanostructure and Nanomaterials: Synthesis, Properties and subramanya, R. H. Colloids Surf. B Biointerf., 2006, 53, 55.
applications,Imperial college press, London, 2004. [36] Pandey, G.; Srivastava, S. K. Synth. React. Inorg. Metal Org. Nano Metal
[7] Sastry, M.; Ahmad, A.; Khan, M. I.; Kumar, R. Curr. Sci., 2003, 85, 162. Chem., 2006, 36, 663.
[8] Perez-Juste J.; Pastoriza-Santos I.; Liz-Marzan L.M.; Mulvaney P. Coord. [37] Hocking, A. D.; Andrews, S. Trans. Br. Mycol. Soc., 1987, 89, 239.
Chem. Rev., 2005, 249, 1870. [38] Seifert, K. Agriculture Agri. Food Canada, 1996, 1.
[9] Gardea-Torresdey, J.L.; Parsons, J.G.; Gomez, E.; Peralta-Videa, J.; Troiani, [39] Li, Y.; Leung, P.; Yao, L.; Song, Q. W.; Newton, E. J. Hosp. Infect., 2006,
H. E.; Santiago, P.; Jose –Yacaman, M. Nano lett., 2002, 2 , 397. 62, 58.
[10] Gardea-Torresdey, J.L.; Gomez, E.; Peralta-Videa, J. R.; Parsons, J.G.; [40] Magaldi, S.; Mata-Essayag, C.; de Capriles, H.; Perez, C.; Collela, M.T.;
Troiani, H.; Jose-Yacaman, M. Langmuir, 2003, 19, 1357. Olaizola, C. Int. J. Infect. Dis., 2004, 8, 39.
[11] Shivshankar, S.S.; Absar, A.; Pasrichaa, R.; Sastry, M. J. Mater. Chem., [41] Mulvaney, P. Langmuir, 1996, 12, 788.
2003, 13, 1822. [42] Slawson, R. M.; Van Dyke, M. I.; Lee, H.; Trevors, J. T. Plasmid, 1992, 27,
[12] Chandran, S. P.; Chaudhary, M.; Pasricha, R.; Ahmad, A.; Sastry, M. Bio- 72.
technol. Prog., 2006, 22, 577. [43] Zhao, G. J.; Stevens, S. E. Biometals, 1998, 11, 27.
[13] Huang, J.; Li, Q.; Sun, D.; Lu, Y.; Su, Y.; Yang, X.; Wang, H.; Wang, Y.; [44] Herrera, M.; Carrion, P.; Baca, P.; Liebana, J.; Castillo, A. Microbios, 2001,
Shao, W.; He, N.; Hong, J.; Chen, C. Nanotechnology, 2007, 18, 104. 104, 141.
[45] Yoshida, K.; Tanagawa, M.; Mastumoto, S.; Yamada, T.; Atsuta, M. Eur. J.
[14] Mandal, D.; Mark E. Bolander, M. E.; Mukhopadhyay, D.; Sarkar, G.; Muk-
Oral Sci., 1999, 107, 290.
herjee, P. Appl. Microbiol. Biotechnol., 2006, 69, 485.
[46] Sondi, I.; Sondi, B. S. Colloids Interface Sci., 2004, 275, 177.
[15] Klaus, T.; Joerger, R.; Olsson, E.; Granqvist, C. G. Proc. Natl. Acad. Sci.
[47] Kim, J. S.; Kuk, E.; Yu, K. N.; Kim, J. H.; Park, S. J.; Lee, H. J.; Kim, S. H.;
USA, 1995, 96, 13611–13614.
Park, Y. K.; Park, Y. H.; Hwang, C. Y.; Kim, Y. K.; Lee, Y. S.; Jeong, D. H.;
[16] Frankel, R. B.; Blakemore, R. P. Iron Biominerals. Plenum: New York, Cho, M. H. Nanomedicine, 2007, 3, 95.
1991. [48] Shahverdi, A.H.; Minaeian, S.; Shahverdi, H.R.; Jamalifar, H.; Nohi, A.A.
[17] Holmes, J. D.; Smith, P. R.; Evans-Gowing, R.; Richardson, D. J.; Russell, Process Chem., 2007, 42, 919.
D. A.; Sodeau, J. R. Arch. Microbiol., 1995, 163, 143. [49] Jeon, H. J.; Yi, S. C.; Oh, S. G. Biomaterials, 2003, 24, 4921.
[18] Husseiny, M.I.; Ei-Aziz, M.A.; Badr, Y.; mahmoud, M.A. Spectrochim. Acta [50] M.R. initiated, designed, and coordinated the project. A.I. and A.G carried
A, 2007, 67, 1003. out most experiments and the data analysis in the lab and under the guidance
[19] Mann, S. Nature, 1995, 365, 499. of M.R. Electron microscopy was performed by S.P. in the lab of C.S. The
[20] Nair, B.; Pradeep, T. Cryst. Growth Des., 2002, 2, 293. manuscript was prepared by M.R with minor input from C.S and S.P.
[21] Sushil, K. S.; Mamta, P. Rev. Adv. Mater. Sci., 2003, 5, 501.

Received: November 11, 2007 Revised: December 15, 2007 Accepted: January 18, 2008

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