Aloe Vera and Neem as

Antimicrobial Agents

By:
Ms. S. K Vyas,
Ms. S. V. Ingale,
Dr. S. Mukhopadhyay
& Mr. N. Saraf
 Aloe Vera and Neem as Antimicrobial Agents
By: Ms. S. K Vyas, Ms. S. V. Ingale, Dr. S. Mukhopadhyay & Mr. N. Saraf

Introduction

In the world of textile and garment, finishing plays a vital role for quality and value. In the abundance of
various finishes, importance is given to antimicrobial finish since people take much care about health and
hygiene. Hygiene has acquired more importance in recent years. Natural fibres are more susceptible to
bacterial attack than synthetic fibres due to their porous and hydrophilic nature. The structure of natural
fibers retains water and oxygen along with nutrients, thereby offering optimal enrichment culture for rapid
multiplication of micro-organisms. On the other hand, direct contact with human body supplies warmth,
humidity and nutrients, i.e., provides a perfect environment and optimal conditions for bacterial growth. The
most troublesome organisms are fungi and bacteria. Under very moist conditions algae can also grow on
textiles. Fungi creates multiple problems to textile including discoloration, stains and fiber damage,
unpleasant odor and a slick, slimy feel. Besides, structure of the substrate and the chemical processes may
induce growth of microbes. Humid and warm environment further aggravates the problem.

The present research work is aimed at developing an eco-friendly antimicrobial finish from neem and aloe
vera plant for textile application. The neem and aloe vera extracts are applied on cotton and silk fabric and a
study is conducted to assess the antimicrobial activity of the finished samples.

What are antimicrobial Finishes?

Antimicrobial agents destroy or inhibit the growth of microorganism. Antimicrobial finishes control, destroy or
suppress the growth of microorganisms and their negative effects of odour, staining and deterioration.
Antimicrobial agents migrate and chemically enter or react with micro-organism. Antimicrobial finishes acts
as a poison for them. There are different types antimicrobial finishes based on chemical and natural
sources.

In recent years, great attention has been devoted to biopolymers because of their biocompatibility and
biological functions and consequently, they are used in textile, biomedical and pharmaceutical fields. Natural
fibres i.e. cotton, silk etc., provide an ecologically friendly textile, but more than 75% of the natural fibers are
finished with synthetic chemicals. Unfortunately, these finishes are unfavorable from an ecological point of
view, because effluents generated are heavily colored, contain high concentrations of salts, and exhibit high
biological oxygen demand/chemical oxygen demand (BOD/COD) values.

Natural Antimicrobial Finishes

Natural finishes are those in which various materials from plant or animal kingdom are used. Some marine
animals such as prawns and fishes posses some compounds which exhibit antimicrobial activity. Chitosan is
an effective natural antimicrobial agent derived from Chitin. Natural herbal products such as neem, tulsi,
pomegranate, aloe vera, prickly chaff flower, turmeric, clove etc also exhibit antimicrobial activity. Studies
reveal that some specific species of herbs having antimicrobial activity are suitable for textile application.

Experimental Work:

Materials

The cotton and silk fabrics of following specifications (Table 1) were taken for the experimentation.

Table 1-Fabric Specification

Sr. No. Specifications Cotton Silk

1 EPI 52 100
2 PPI 48 52
3 GSM 190 85
4 Weave Plain Plain
Antimicrobial agents like neem and aloe vera were used. These are natural plants which have very good
antimicrobial effect together with other functional properties suitable for textiles.

Pretreatment of Fabrics:

• The cotton fabric was scoured by caustic boil method.

• The scoured fabric was bleached by calcium hypochlorite bleaching process.

Extraction Processes:

Aloe Gel Extraction

Careful processing of aloe gel is necessary to maintain activity. Aloe was

processed using hand filleted or whole leaf procedure. This hand filleted
processing carefully removes the inner gel while avoiding the yellow sap
(latex) found next to the rind. The latex contains aloin, which is removed for
internal and external gel products. Stabilizing the constituents within aloe is
another essential part of developing effective products. The gel must be
processed under cold conditions within 4 hours of harvesting. The gel was
smashed and converted into solvent form.

Neem Extraction

The fresh leaves of neem were taken and shadow dried. After complete drying they were made into a fine
powder by grinding and filtering. The extracted neem powder was treated with methanol at room
temperature, resulting in active substances which were subsequently dissolved in methanol. The herbal
materials were mixed with methanol in the ratio 1:5 in a beaker. The beaker was left closed for 3 days so
that the methanol takes in the active components of the leaves. After 3 days, the methanol was let to
evaporate, so that the residue gets settled at the bottom of the beaker.

Finish Application:

The antimicrobial agents can be applied to the textile substrates by exhaust, pad-dry-cure, coating, spray
and foam techniques. Here exhaust method was used. The processed gel and neem extracts were treated
acetic acid as mordant in the ratio 1:10. The liquor was finished onto the cotton and silk fabric in various
concentration and combination (50:50) at following process conditions:

Temperature - 800C
pH -5
Time - 30 min.
M : L Ratio - 1:20

The fabric was then dried at 80°C for 15 minutes to remove the moisture.

The various concentrations were as

NEEM - 3% and 5%
ALOE-VERA - 3% and 5%
ALOE-VERA + NEEM - 3% and 5%

Table 2 Nomenclature of samples

Sr. No. Ingredient and concentration Nomenclature

1 Untreated Sample Untreated
2 3% Neem Neem3
3 5% Neem Neem5
4 3% Aloe vera Aloe3
5 5% Aloe vera Aloe5
6 3% (Aloe vera + Neem) A3+N3
7 5% (Aloe vera + Neem) A5+N5
Agar diffusion Test:

This test is used for the qualitative and quantitative assessment of antimicrobial finishes on the textile
material. It measures the degree of antimicrobial activity on the textile material.

Quantitative Assessment of Antibacterial finishes on textiles measures the degree of antibacterial

activity (AATCC-100-1998):

Fabric samples of size 2x2 inches were taken from cotton and silk fabric. Fabric samples were treated with
neem, aloe vera and their combination (neem + aloe vera) with 3% and 5% concentrations. 500ml conical
flasks containing 50ml of nutrients broth were prepared and sterilized at 1210C for 15 minutes. It was then
allowed to cool. The fabric samples were subsequently transferred aseptically into conical flasks
respectively. These were incubated at 370C for 24 hours in shaker at 121rpm. After incubation samples were
tested for absorbency values on spectrophotometer at a wavelength of 600 nm and are used to know the
effectiveness of antimicrobial treatment.

Qualitative Antibacterial Assessment of diffusible antibacterial agents ("quick method") (AATCC-

147-1998):

In this test method 50ml of nutrient agar was prepared and sterilized at 1200C for 15 minutes. Petri plates
were autoclaved in hot air oven at 1210C for 30 minutes. 20ml of nutrient agar was poured into each of these
plates and were allowed to solidify. A series of 8 test tubes containing 4.5ml of sterile water was taken. 0.5ml
of culture from nutrient broth containing the 3% gel treated sample was transferred aseptically into the first
test tube (As shown in fig. 2). Serial dilution was carried out until its reduced dilution was 10(-8) micro liters.
10(-8) diluted culture was taken aseptically and poured onto the petri plates. This was spread by using L rod.
The plates were incubated at 370C for 16-18 hours. After incubation these Petri plates were observed for the
number of colonies were developed. Similar procedure was carried out for all untreated sample and treated
samples.

Fig. 2 Quick method

Assessment of Durability of Finish to Washing:

The durability of the finish to washing is analyzed by testing all finished samples on the 'Digi-wash' tester. In
this instrument fabric samples are subjected to a controlled and predetermined number of washing cycles.

At first the finished fabrics are cut into samples of 100mm × 25 mm and a soap solution is prepared by
taking the following materials-

Standard soap (powder form) - 4 gm/l

Sodium carbonate (Na2CO3) - 2gm/l

These are mixed with water by taking the M:L ratio 1:20. A homogeneous soap solution is obtained by
stirring the solution for few minutes. The fabric samples and soap solutions were taken together in the steel
cups of the 'Digi-wash' tester. The cups were fastened inside the instrument and the lid was closed. The time
was set at 10 minutes and temperature of 400C. After the completion of washing the samples were washed,
dried, and again subjected to a desired number of washing cycles (5, 10, 15, 20 and 25 cycles) in a similar
way explained above. The fabric samples were then subjected to microbial agar and the bacterial growth
was analyzed by serial dilution carried out after incubation.

Results and Disccusson:

Quantitative Assessment of Antibacterial finishes on textiles measures the degree of antibacterial

activity (Performed according to AATCC-100-1998):

Fig.3 shows two conical flasks which contain

treated and untreated fabric samples after
incubation (shown by arrows). The turbidity of
both the flasks is checked on spectrophotometer
and absorbance values are recorded.
Absorbance of the sample is directly proportional
to the concentration of the cells in the sample.
Hence the sample, which gives lower absorption
value, has more microbial resistance and vice-
versa.

Absorbance test (600nm)

The absorbance values for all 12 samples are

given in Table 3 and Table 4. The direct comparison of all the values can be easily seen in the graphs
plotted below. From these graphs we can easily understand the level of antimicrobial resistance of all the
finished samples.

Untreated Sample of cotton = 1.367

Table 3- Bacterial Absorbance values of treated samples for cotton

Sample Absorbance Value Sample Absorbance Value

Aloe 3 1.112 Aloe 5 1.020

Neem 3 1.022 Neem 5 0.687

A3+N 3 0.796 A5+N 5 0.334

Fig. 4 Absorbance values for cotton (treated and untreated)

Untreated sample of silk = 1.254

Table 4- Bacterial Absorbance values of treated samples for silk

Sample Absorbance Value Sample Absorbance Value

ALOE 3 1.164 ALOE 5 1.019

NEEM 3 1.096 NEEM 5 0.852

A3+N 3 0.895 A5+N 5 0.373

Fig. 5 Absorbance values for silk( treated and untreated)

The absorbance values for all 12 samples are given in Table 2 and Table 3. When cotton and silk samples
were treated with aloe vera i.e. 3% & 5% concentration and combination, the resultant fabric do not support
the growth of bacteria compared to untreated fabric. Samples were treated with 3%, 5% concentration of
neem not only gave low absorbance value as compared to untreated sample but also showed superior
values than aloe vera treated samples. This indicates that neem resist the growth of bacteria to a higher
extent. When samples were treated with combination of neem and aloe-vera (50:50) each with 5%
concentration gave lowest absorbent values which show that these samples resist growth of bacteria to the
highest extent. It also shows that cotton gave lower values than that of silk.

Qualitative Antibacterial Assessment of

diffusible antibacterial agents (“quick
method”) (Performed according to AATCC-
147-1998):

Figure 6 shows the view of petri plates after

incubation. After incubation, the petri plates
were observed for bacterial growth. Then
colonies were counted for each plate. Samples
treated with 3% and 5% concentration of neem
got less number of colonies than Aloe vera
treated samples. This indicates that neem
treated fabric do not support bacterial growth,
whereas aloe vera samples shows smaller
extent of resistance to microbial growth.
Samples treated with combination of aloe vera and neem with each 5% concentration had less number of
colonies when compared to other samples.
Bacterial observation of finished samples with respect to washing cycles:

The results obtained from the serial dilution of the samples from washing fastness test are as shown in the
Table 5.

Table No. 5 Washing fastness for different fabric/ Antimicrobial agent combination

Sample Cotton Cotton Cotton Silk Silk Silk

Antimicrobial Aloe Neem Combination Aloe Neem Combination
Agent vera (A+N) vera (A+N)

5 washings -- -- -- -- -- --
10 washings -- -- -- -- -- --
15 washings -- -- -- -- -- --
20 washings -- 2 1 -- 2 --
25 washings 1 3 2 1 3 2

It was observed that that there was no bacterial growth in the finished samples up to 15 washings. After 15
washings there was less bacteria observed in neem finished cotton and silk samples and in cotton sample
finished with both the antimicrobial agents in combination. The cotton and silk samples finished with aloe
vera shows no bacterial growth even after 20 washings. After 25 washing cycles all samples shows some
bacterial colonies. Aloe vera samples show lowest level of bacterial growth. Hence, it is obvious that aloe
vera finish has the best durability for washing. Second comes the combination finish and samples finished
with neem shows comparatively a lower level of durability for washing.

Conclusions:

The present study supports the potential and viability of natural agents as antimicrobial finish. The extracted
materials have good antimicrobial properties.

From the present study following conclusions are derived–

• Natural antimicrobial agents used viz., Neem and Aloe vera have been effective on cotton and silk
fabric.
• These natural antimicrobial agents provide good protection when used in combination.
• Neem shows more microbial resistance than aloe vera.
• For both cotton and silk a combination of neem and aloe vera in a ratio of 50:50 using a 5%
concentration for each ingredient shows the best antimicrobial property.
• The antimicrobial effect on the fabric samples is durable even after 15 washes. Aloe vera when used
as a single component fared the best even in comparison to the combination of aloe vera + neem in
term of durability.
• Since the active ingredients are extracted from nature, this methodology can serve as a suitable
alternative to conventional harmful chemicals in present use.

References:

1) G Thilagavathi & Rajendrakumar, "Development of eco-friendly antimicrobial textile finishes using

herbs" Indian Journal of Fibre & Textile Research Vol. 30, December 2005
2) G. Raj Kumar and V. D Saravanan "An Eco Friendly Herbal Antimicrobial Finish on Cotton Fabric
Using Aloe Barbadensis Miller (Aloe Vera)", The Indian Textile Journal, October 2005
3) Dr G Thilagavathi, K Rajendrakumar & R Rajendran "Development of antimicrobial textile finishes
from plant species" www.expresstextile.com
4) Barrie Clemo (TRA) "Antimicrobial concepts to enhance textile value chain", Indian Textile Journal,
July 2004
5) Dr Shanmugasundaram "Antimicrobial finish in textiles" www.indiantextilejournal.com
6) B. Ramesh Babu, A.K. Parande, S. Raghu, and T. Prem Kumar "Cotton Textile Processing: Waste
Generation and Effluent Treatment" The Journal of Cotton Science 11:141-153 (2007)
http://journal.cotton.org