Volume 7, Issue 2, February – 2022 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Evaluation of Antibacterial and Antifungal

Susceptibility of Water Hyacinth
Dr A Vaidya Soocheta; Sr. Lecturer, Faculty of Engineering, University of Mauritius, Réduit, Mauritius; [email protected]
J Jadheea; MPhil/PhD Student, Faculty of Engineering, University of Mauritius, Réduit, Mauritius;
Dr M Nerurkar; Managing Director, Calantha Biotech Pvt. Ltd. Saki Vihar Rd, Mumbai 400072;

Abstract:- Water hyacinth (Eichhornia crassipes) is an may lead to the development of antimicrobial drugs,
aquatic weed which proliferates rapidly in lakes and cosmetics products including soaps (Cowan, 1999). Several
rivers. The rapid growth and spread of water hyacinth medicinal plants have been studied in the field of
has been a great concern worldwide. In Mauritius, it pharmaceutical and scientific community (Baral and Vaidya,
covers an approximate area of 13.67 km² throughout the 2011). Baral and Vaidya, 2011 also state that many
island. The biodegradability, antibacterial and antifungal researchers are exploring the potential of plants for their
activity of water hyacinth’s leaves, stalks and roots were antimicrobial usefulness. Since centuries, herbs have been
studied to explore it’s potential as a raw material for used as the main source of medicinal drugs (Sa’ad et al.,
product development. The inoculums Trichoderma viride, 2005). According Shanab et al., 2010, water hyacinth has a
Staphylococcus aureus, Klebsiella pneumoniae and high chemical potential for medicinal functions. The leaves
Aspergillus niger were used for testing. The of the weed contain phenols, alkaloids, tannins, flavonoids
biodegradability test was conducted with enzyme which contribute as antibacterial agents and to antiviral,
cellulase from Trichoderma viride using the enzymatic antifungal, antitumor activities. According to Haggag et al.
method. The biodegradation levels were determined by (2017), the leaf extracts and compounds of water hyacinth
computing the percentage weight loss after thirty days. show anti-microbial and antifungal activities. Phenolics and
The antibacterial test was performed with the test antioxidants are secondary metabolites that are considered as
inoculums Staphylococcus aureus and Klebsiella chemical defence of plants against plant pathogens (Haggag
pneumoniae. Aspergillus niger derived from ATCC 6275 et al., 2017). The weed contains a substantial amount of
was used for the antifungal test. The results of the tests oxidative enzymes and non-enzymatic antioxidant systems
provide promising prospect for advancing into the (Thamaraiselvi et al., 2012). Researchers need to show more
development of ecologically friendly products. interest in common weeds which are rich in phytochemicals
(Jayanthi and Lalitha, 2013). Antibacterial activity against
Keywords:- Water hyacinth, Biodegradability, Antibacterial, Staphylococcus albus and antifungal activity against Mucor
Antifungal. sp that has been studied by Jayanthi and Lalitha (2013),
demonstrates that water hyacinth can be a potential source for
I. INTRODUCTION developing antimicrobial agent that can be used in cosmetic
products.
Water hyacinth also known as Eichhornia crassipes is a
highly invasive aquatic plant that propagates mostly in II. MATERIALS AND METHODS
tropical and sub-tropical regions. Water hyacinth grows into
dense mats causing severe obstructions in rivers, lakes and A. Sampling:
dams which result in floods, blockages to marine Water hyacinth leaves, stalks and roots used for testing
transportations and turbines. Water hyacinth propagation also were harvested from Haut Rive Lake, at Riviere Du Rempart,
increases eutrophication and cause algal blooms. The weed Mauritius. They were separated and washed thoroughly under
serves as a habitat for many disease vectors such as malaria. running water. They were then dried under the sun. The dried
In several developing countries, the spread of water hyacinth leaves, stalk and leaves were used for the study. These were
is triggering socio-economic problems to communities that finely ground and tested for the biodegradability test and
depend on freshwater bodies. However, literature and studies antibacterial test. A circular fibre swatch of 38 mm was made
also reveals this weed to be a productive plant (Kayathri et with the leaves, stalk and roots, and used for the antifungal
al., 2015). Recently, considerable attention has been given to test.
harvest the plant and put to use to partially defray the cost of
removing it (Lata and Dubey, 2010). This study focuses on B. Biodegradability Test:
the biodegradability, antibacterial properties and antifungal Enzymatic method ASTM D 5988-03 standard was used
activity of water hyacinth to investigate its potential for the to test the biodegradability of the ground samples (leaves,
development of sustainable products. Antimicrobials are stalk, roots). 1 gram of the sample was tested using cellulase
substances that neutralize or inhibit the growth of enzyme Trichoderma viride for a period of 30 days. The
microorganisms such as bacteria, fungi or protozoans through percentage of weight loss was calculated as follows:
micro biocidal or micro biostatic action. Many plant extracts
have a very high antimicrobial potential. The phytochemicals
present in the plants have a lot of therapeutic properties that

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Volume 7, Issue 2, February – 2022 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
Percentage Weight Loss (%): 100 x (W1 – W2) / W1 Y: Number of colonies at 24 hrs

Where, D. Antifungal Test:

W1: Weight of original sample The antifungal test was carried out using the AATCC
W2: Weight of sample after 30 days 30:III-2013 standard method. Each sample was combined to
make a circular fibre swatch of 38 mm. The samples were
C. Antibacterial Test: placed in a humidity chamber (>90% RH) at 28°C and
Water hyacinth’s leaves, stalks and roots were analysed incubated for 6 days. Aspergillus niger (ATCC 6275) was
for their antibacterial activities using ASTM E 2149-13a used as the test inoculation. The number of colonies were
Method. The samples were ground and 1 gram of each sample counted (X) at ‘0’ hours and (Y) after a period of 24 hours.
was placed into a 100ml of inoculating petri dish.
Staphylococcus aureus ATCC 6538 (Gram Positive Bacteria) III. RESULTS AND DISCUSSION
2.00 x 10^5 and Klebsiella pneumoniae ATCC 4352 (Gram
Negative Bacteria) 2.00 x 10^5 were used for inoculation. To A. Biodegradability Test:
assess the number of viable bacteria for each petri dish, a The biodegradability test was conducted by ASTM D
colony forming unit (cfu) was used. Sterilization was 5988-03 standard test using cellulase enzyme Trichoderma
conducted in an autoclave. viride. The percentage weigh loss of each sample was
recorded. The results in Table 1 show the susceptibility of
Percentage Reduction of Microorganisms (M): 100 x (Y – X) water hyacinth biomass to degradation. It was observed that,
/Y for every 1 gram of leaf, stalk and root samples, it takes 30
Where, days to degrade with a weight loss of 99.6%, 99.7% and
M: Reduction of Microorganisms (%) 99.5% respectively.
X: Number of colonies at 0 hr

Table 1: Biodegradability test

Weight of sample after 30 day Percentage of weight loss

Sample Weight of sample (W1)
(W2) (%)

Water hyacinth leaves 1 gm 0.005 gm 99.6

Water hyacinth stalks 1 gm 0.003 gm 99.7
Water hyacinth roots 1 gm 0.004 gm 99.5

B. Antibacterial Test:
Table 2 shows the results of the antibacterial activity of leaves, stalks and roots with ASTM E 2149 standard test. It records
the number of colonies that are seen at ‘0’ hours (X) and the number after ‘24’ hours (Y). The percentage reduction of
microorganisms (M) was calculated. The antibacterial activity of the leaves with test cultures S. aureus and K. pneumoniae was
99.23% and 99.03% respectively. The stalks show an antibacterial activity of 99.87% with S. aureus and 99.70% with K.
pneumoniae. The roots showed 99.88% antibacterial activity with S. aureus and 99.85% K. pneumonia. From the results it can be
inferred that the plant displays good antibacterial properties against S. aureus and K. pneumonia.

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Volume 7, Issue 2, February – 2022 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
Table 2: Antibacterial test
No. of Reduction of
No. of colonies
Sample Test Culture colonies‘24’ Microorganisms (M)
‘0’ hour (X)
hours (Y) %
S.aureus 2.08 X 10^5 1.6 X 10^3 99.23%

K. pneumoniae 2.17 X 10^5 2.1 X 10^3 99.03%

Leaves
S.aureus 2.09 X 10^5 3.7 X 10^2 99.87%

K. pneumoniae 2.11 X 10^5 6.3 X 10^2 99.70%

Stalks
S.aureus 2.05 X 10^5 2.3 X 10^2 99.88%

K. pneumoniae 2.06 X 10^5 2.9 X 10^2 99.85%

Roots

C. Antifungal Test:

Table 3: Material resistance rating

Growth on specimen Rating
No growth 0
Trace of Growth (<10%) 1
Light Growth (10 to 30%) 2
Medium Growth (30 to 60%) 3
Heavy Growth (60% to complete coverage) 4

The percentage of surface growth of Aspergillus niger was rated on a scale of ‘0’ to ‘4’, as seen in Table 3 for the testing the
material resistance rating. ‘No growth’ is indicated by ‘0’ and a heavy growth of 60% to complete coverage is given a rating of ‘4’.

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Volume 7, Issue 2, February – 2022 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
Table 4: Antifungal test
Sample Zone of Inhibition Rating Interpretation
A zone of inhibition was identified around the fibre. There was
Leaves 4.2 mm zone of inhibition 0
no trace of fungal growth on the sample.
A zone of inhibition was identified around the fibre. No trace of
Stalks 4.4 mm zone of inhibition 0
fungal growth was spotted on the sample.
A zone of inhibition was identified around the fibre. No growth
Roots 4.2 mm zone of inhibition 0
of fungal was detected.

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