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Environmental Pollution 69 (1991) 69-78 • .~

Biological Treatment of Wastewater by Selected Aquatic

Plants

B. D . T r i p a t h i * & S u r e s h C. S h u k l a

Pollution Ecology Research Laboratory, Centre of Advanced Study in Botany,

Banaras Hindu University, Varanasi-221005, India

(Received 8 September 1989; revised version received 23 July 1990;

accepted 26 July 1990)

ABSTRACT

Pollutant-removal efficiency of certain macrophytes and algae, such as

Eichhornia crassipes, Microcystis aeruginosa, Scenedesmus falcatus,
Chlorella vulgaris and Chlamydomonas mirabilis, has been tested in
laboratory conditions to evaluate their potential role in wastewater treatment.
Sewage of Varanasi city, mixed with the effluents of about 1200 small-scale
industries, was used for the tests. The investigation was performed in three
stages i.e. a water hyacinth culture Jbllowed by an algal culture, and finally a
second water hyacinth culture. For the first water hyacinth culture, 10 water
hyacinth plants were grown in a tank of wastewater with 15 days' retention
time. In the second stage, algal species were cultured in the treated wastewater
.for 5 days, whilst in the third stage, water hyacinth plants were again grown
./br further treatment of the wastewater .for 9 days. This three-stage
aquaculture resulted in very high reductions of BOD (96"9%), suspended
solids (78"1%), total alkalinity (74.6%), PO4--P (89.2%), N O 3 - - N
(81.7%), acidity (73.3%), N H 4 - - N (95.1%), COD (77.9%), hardness
(68"6%) and coliform bacteria (99"2%). An increase in the concentration of
dissolved oxygen ( 7 0 % ) was also observed.

INTRODUCTION

In recent years, d o m e s t i c sewage a n d industrial effluents h a v e been r e g a r d e d

as a c o m m o n s o u r c e o f pollution. A t Varanasi, a b o u t 126 m 3 d - 1 u n t r e a t e d
* To whom correspondence should be addressed.
69
Environ. Pollut. 0269-7491/91/£03"50 © 1991 Elsevier Science Publishers Ltd, England.
Printed in Great Britain
70 B. D. Tripathi, Suresh C. Shukla

domestic sewage mixed with industrial effluents is discharged into the river
Ganga. In the discharge areas, microbial decomposition of these substances
exerts high BOD, creates anaerobic conditions and the receiving aquatic
bodies become septic and emit foul smells. Under these conditions aquatic
life suffers, resulting in a loss of productivity of the natural waters and a
deterioration of water quality to such an extent that the water becomes
unusable. Therefore, proper treatment of the sewage is urgently needed.
The use of aquatic macrophytes to remove nutrients from sewage and
industrial effluents has been suggested by Boyd (1968), Wolverton &
McDonald (1979), Paverly (1983), Oron et al. (1986), Sutton & Ornes (1975),
Reddy (1983), Shukla & Tripathi (1989) and Tripathi et al. (1990). Also, in
recent years algae have been used extensively in treatment systems
(Christensen et al., 1979; Shelef et al., 1980; Rodrignes & Oliveira, 1987;
Tam & Wong, 1989).
Aquatic macrophytes can be effectively used to reduce pollutant levels in
water bodies (Boyd, 1968; Lakshman, 1979; Stowell et al., 1981; Reddy et al.,
1983; Hauser, 1984; Reddy & De Busk, 1985; De Busk et al., 1989) and their
biomass may be used for biogas production (Shialipour & Smith, 1984)
animal feed (Bagnall et al., 1974), fibre (Nolan & Kirmse, 1974) and compost
(Parra & Hortenstein, 1974). The use of water hyacinth not only helps in
BOD depletion, but also reduces suspended solids, NO3--N, PO4--P, Na,
K, Ca, Mg and other minerals (Tripathi et al., 1990). This plant can absorb toxic
substances, such as cadmium, lead, nickel, mercury, copper, chromium,
silver, phenol and some potential carcinogens (Wolverton & McDonald,
1979). These aquatic plants can absorb these elements at concentrations
between 4 000 and 20 000 times those in water (Wolverton & McDonald,
1979). The effect of nutrient availability on water hyacinth standing crop and
detritus deposition have been studied by Thomas et aL, 1989.
However, most of the past studies were restricted to one kind of aquatic
macrophyte or alga for wastewater treatment. Therefore, a systematic
sewage treatment technique was designed to evaluate the potential of
combining water hyacinth and algae (Microcystis aeruginosa, Seenedesmus
quadricauda, Chlorella vulgaris, Euglena viridis) in improving water quality.

MATERIALS AND METHODS

Wastewater of Varanasi city (city sewage mixed with industrial effluents)

was collected every day between 06.00 h and 07.00 h from Rajghat nala,
brought to the laboratory and mixed by mechanical agitators. After
screening, the homogenised wastewater was transferred to a 35-1itre plastic
 Biological treatment of wastewater by selected aquatic plants 71

STAGE-I STAGE-II STAGE-III

-_--_-- -- -- --_~ ~
-_- --_- -_- -_-
-_--___=--__--__. _-_~ I
-----___-_--_.
7

TREATED
WATER
]--_---_----:_-S-~_-
t -J
ALGAL C U L T U R E EICHHORNIA,-
CRA URE-I CRASSIPES CULTURE-II

Fig. 1. Biological treatment of wastewater by Eichhornia crassipes and algal culture.

container and allowed to settle for 2 h. After sedimentation, the wastewater

was transferred to a 20-1itre aspirator bottle and allowed to flow through the
experimental set-up shown in Fig. 1.

Stage I treatment

The wastewater from the aspirator bottle was allowed to flow into the
bottom of the first water hyacinth culture tank through a 1"25-cm pipe
regulated by an adjustable valve. The water-holding capacity of tank was
200 litres. Ten water hyacinth plants were grown in the wastewater as cover
crop and kept exposed to sunlight. Regular harvesting of the older plants
was done once a week, so that 15-20% of the water surface in the tank
remained free from the plants. The retention period for this water hyacinth
culture was 15d.

Stage II treatment

Treated wastewater from the first water hyacinth culture tank was allowed
to overflow, through an outlet pipe fixed 8 cm below the top of the tank, into
an algal culture tank. The capacity of this algal tank was about 60 litres.
Algae were allowed to grow for 5 days in the partly treated wastewater
received from the first water hyacinth culture.

Stage III treatment

After 5 days, water from the algal culture tank was again allowed to overflow
into the second water hyacinth culture tank-II with a capacity of 200 litres.
72 B. D. Tripathi, Suresh C. Shukla

At this stage, water hyacinth was again grown as a cover crop for a further 9
days. The treated water from this second water hyacinth culture was
discharged as purified water.
Water samples were collected every morning from the outlet of each
treatment tank. All the determinations, i.e. suspended solids, total alkalinity,
BOD, COD, EC, acidity, NO3--N, PO4--P, total hardness, cadmium,
coliform bacteria, phytoplankton and zooplankton density were done
following the methods descrived in APHA (1985). Due to high variation, the
pH and dissolved oxygen (DO) were determined twice a day at 09.00 h and
13.00 h. DO was determined by a Century automatic 0 2 analyser. An atomic
absorption spectrophotometer (model IL 751) was used to determine the
cadmium, copper, iron, nickel, zinc and lead concentrations in the water
samples.
Phytoplankton and zooplankton populations in the water were analysed
at three different stages in the aquaculture. The phytoplankton density was
estimated by the microtransect method described by Lackey (1938) and
subsequently modified by Edmondson (1974). Zooplankton were counted
by the method described by Jhingran et aL (1969), and coliform bacteria were
estimated by the presumptive and confirmatory test using MacConkey's
broth (APHA, 1985). The statistical significance of variation in the data with
respect to days for different parameters was judged with the help of the 'F'
(Variance ratio) test.

RESULTS A N D DISCUSSION

The average data obtained over a 6-month experimental period are given in
Table 1.

Stage I (water hyacinth culture-l)

All the physicochemical and bacteriological parameters, such as suspended

solids, pH, total alkalinity, biochemical oxygen demand (BOD), electrical
conductance, chemical oxygen demand (COD), acidity, NO3--N, PO4--P,
total hardness and coliform bacteria count were reduced and water quality
improved in the first water hyacinth culture. During a 15 days' retention
period, suspended solids were reduced from 320 mg litre- 1 to 135 mg litre-
and BOD from 310 mg litre- ~ to 70 mg litre- 1. Significant reductions in pH,
total alkalinity, electrical conductance, COD, acidity, NO3--N, NH4--N,
PO4--P, total hardness and coliform bacteria were observed. The pH and
DO concentrations were recorded at 09.00 h and 13.00 h. The pH varied
from 6.45 to 7.69, and the DO concentrations varied from 0.64 to 1.4 mg
 Biological treatment of wastewater by selected aquatic plants 73

v~

+1+1+1+1+1 +1+1+1+1+1+1+1+1 +1+1 x ~

=_..

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~ ° ° ° ~ ~ ~ ° ~ ° ~ ~ +1
+1+1+1+1+1 +1+1+1+1+1+1+1+t ~1+1 x - x~
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I

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74 B. D. Tripathi, Suresh C. Shukla

litre- 1. A low density ofphytoplankton was recorded (775 x 10 3 individuals

litre-1), due probably to the water hyacinth restricting the penetration of
sunlight, which inhibited the growth of these algae. Z o o p l a n k t o n
populations were only 290 individuals litre -1 in the first water hyacinth
culture. A depletion in the cadmium concentration, from 1.6 mg litre-1 to
0-96mg litre -1, was also observed in this treatment. The wastewater
obtained after the first stage treatment was free from any odours.

Stage II (algal culture)

The water quality from the first water hyacinth culture was quite good with
respect to BOD, suspended solids, PO4--P, NO3--N, N H 4 - - N , COD
and coliform bacteria count. However, the DO concentration was not high
enough to constitute good quality water. For this reason, algae, such as
Chlorella vulgar&, Microcyst& aerug&osa, Scenedesmus falcatus and
Chlamydomonas mirabilis, were cultured in this second stage in order to
increase the concentration of DO through their photosynthetic activities.
During 5 days' retention time of the algal culture, BOD was further
reduced from 70-0 mg litre- 1 to 40.5 mg litre - 1, total alkalinity from 470 mg
litre-1 to 290 mg litre-1, PO4__ P from 4.65 to 3"05 mg litre-1, NH4__ N
from 39-4 to 16.5 mg litre- 1, COD from 567 to 450mg litre- 1, EC from 850
to 775#S, total hardness from 64 to 49.5mg litre -1 and total coliform
bacterial count from 2.14 x 105 to 0-76 x 155 cells 100m1-1. Calcium
concentration was further reduced from 0.96 to 0.68 mg litre-1
Thus the incorporation of the algal culture after the first water hyacinth
culture resulted in an overall improvement of water quality. However, the
growth and multiplication of algae increased the suspended solids from
135 mg litre- 1 to 205 mg litre- 1. The pH was 7.25 at 09"00 h, but it was 9.04
at 13"00h. This increase in the pH at 13-00h was possibly due to active
photosynthetic activity during the day, resulting in a reduction of CO 2 in the
water. The DO content of the water at 09.00h was 3"64mglitre-~, but it
increased at 13.00h to 14-5mglitre-1 due to photosynthetic activity of
algae. Oxygen remains in the water in supersaturation state. The
p h y t o p l a n k t o n population increased from 775 × 10 3 to 1598 × 104
individuals litre-1. Similarly, the zooplankton population increased from
210 to 6759 individuals litre-1.
Thus, the outlet water from the algal culture was much improved with
respect to BOD, NO3--N, PO4--P, N H 4 - - N , pH, DO, COD, hardness,
electrical conductance, coliform bacterial count and zooplankton numbers.
However, the concentration of suspended solids increased due to the
presence of algal cells and, as a result, the water was unfit for discharge,
Hence, a further water hyacinth culture-II was used to reduce the suspended
solids before discharge of treated wastewater.
 Biological treatment of wastewater by selected aquatic plants 75

Stage-Ill (water hyacinth culture-lI)

The separation of suspended algal cells from treated water is very difficult.
The processes by which algae may be removed include dissolved air
flotation, centrifugation, and pressure filtration through microstrainers. All
of these techniques are expensive and have seldom been used on a
commercial scale.
The outlet water of stage II was allowed to flow through the second water
hyacinth culture. The water hyacinth plants growing in this culture were
luxuriant, dark green and healthy, and during this stage all of the pollutants
were further reduced, yielding good water quality at discharge.
Quantitative analysis at this stage, revealed 9.65mglitre-1 of BOD,
70.5 mg litre- 1 suspended solids, 170.5 mg litre- 1 total alkalinity, l ' 1 4 m g
litre- 1 PO4--P, 0"30 mg litre - 1 NOa__N, 3.14 mg litre- ~ N H 4 - - N , 36-4 mg
litre -1 total hardness, 0"114 × 105 cells 100ml -~ coliform bacteria and
0.4mglitre-1 to 12.1mglitre-I dissolved oxygen. The cadmium con-
centration was reduced to 0-27 mg litre -1.
A satisfactory reduction in phytoplankton density from 1598 × 104 to
1235 × 1 0 3 individuals litre-~ was also observed. However, an increase in
the zooplankton population from 6759 to 10946 individuals litre -~ was
found. Coliform bacteria were further reduced from 0.76 x 105 to 0.114 x
105 cells 100ml- 1. Analysis of variance indicated significant (p < 0-001)
differences in all the physicochemical and biological parameters with respect
to time (Table 1).
During the present investigation it was observed that a significant re-
duction in sewage N O 3 - - N was recorded only after a reduction of NH4--N.
This is possibly due to utilization of N H 4 - - N by the algae, so leaving nitrite
and nitrate in the wastewater. Chevalier and Noue (1985) observed that
more nutrients were utilized by algal cells in hyperconcentration conditions,
which indicates effective removal of nutrients during wastewater treatment.
Water hyacinth removes many of the constituents of wastes by physical
absorption into its tissues, where bioaccumulation can be as high as 20 000
times (Aowal & Singh, 1981). During the absorption of water, colloidal
particles in the wastes continuously impinge on the surface of the hairy roots
of water hyacinth and become agglomerated after losing their electrical
charges. The agglomerated particles carry with them more colloidal
particles, as they settle, and the phenomenon seems to be more physical than
chemical. Reduction in the coliform count by water hyacinth is also
significant, and it may be that the agglomeration of colloidal particles in the
wastewater carries the bacteria to the bottom.
Coagulation of colloidal and suspended particles in the wastewater can
occur by various means, such as addition of electrolyte, evaporation or
cooling (Glasstone, 1965), so the coagulation of colloidal particles may be
76 B. D. Tripathi, Suresh C. Shukla

due to absorption of the liquid through the surface of roots of the plant. It is
also possible that the roots may be absorbing some materials which act as
peptising agents for the colloids. The removal of biodegradable organics is
also accomplished by the symbiotic activity of a large number of aerobic
bacteria that grow on the roots of the plant. These aerobic bacteria get their
required oxygen through the vascular stem of the plant.
The present experiment, conducted with water hyacinth and a few algal
species (Chlorella vulgaris, Scenedesmus quadricauda, Microcystis aerugi-
nosa, Euglena viridis) revealed that the use of aquatic plants for the treatment
of sewage is a cheap and convenient technology. Controlled growth of plants
can produce cost-effective, advanced wastewater treatment systems in warm
and temperate climates. It is suggested that it will be more economic and
desirable to employ such techniques on commercial scale. Plant material
harvested from this type of system can be further processed into usable
products like compost, paper and other products. Anaerobic digestion of the
harvested biomass produced (60-80%), 374 litres methane kg-1 dry plant.
One hectare can produce 600 kg of dry plant material per day, which can
produce 224 400 litres of biogas hectare-1 d-1. With metal-contaminated
plants, the rate of production of gas is even faster (Haque & Sharma, 1986).
When the water hyacinth plants contain heavy metals, such as Ni and Cd,
the methane content in the biogas goes up by 91.9%.
The three-stage system of wastewater treatment described is probably the
cheapest and most economic method which can be adopted throughout
warmer and temperate climates.

ACKNOWLEDGEMENTS

The authors are grateful to Professor R.S. Ambasht, Head, Botany

Department, Banaras Hindu University, Varanasi for providing the
necessary facilities. Thanks are also due to Professors H. D. Kumar and J. S.
Singh for their valuable suggestions and the Ministry of Environment and
Forest, Department of Environment, New Delhi, Govt of India for financial
support.

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