Bioresource Technology 37 (1991) 211-214

Biogas Production from the Aquatic Weed Pistia

( Pistia stratiotes )
S. A. Abbasi, P. C. Nipaney
Salim Ali School of Ecology, Pondicherry (Central) University, Pondicherry 605 001, India

&
M. B. Panholzer
Institute for Environmental Research, Elisabethstrasse 11, A-8010 Graz, Austria
(Received 8 April 1990; revised version received 20 October 1990; accepted 30 October 1990)

Abstract
Pistia stratiotes, an aquatic weed, was investigated
as a substrate for biogas production in batch digestion. An inoculum was necessary to obtain biogas
production from the weed. With Pistia only, production of carbon dioxide alone was high during
the first five days of digestion but began to level off
thereafter With inoculated Pistia, a high rate of
biogas production was sustained for nearly 10 days
and the average methane content was 58-68%.
The digesters charged with Pistia alone had significant concentrations of propionic, butyric, isobutyric, valeric, and isovaleric acids. These acids
were not present in detectable concentrations, in
the digesters running with inoculated Pistia, except
during the first 4 days of the digestion when
propionic acid was formed.
When an inoculum was added to a 'soured'
digester the performance of the latter improved
dramatically.

Key words: Aquatic weed, anaerobic digestion,

biogas, batch digestion, Pistia, volatile and fatty
acids, methane.
INTRODUCTION

Pistia stratiotes L., commonly known as water

lettuce or water cabbage, is a pleustonic macrophyte, originally from the states of Florida and
Texas, USA (Bua-Ngam & Mercado, 1975;
Attionu, 1976; Rao & Reddy, 1984; Tadulingam

& Venkatanarayana, 1985). It is now widespread

in the lakes and ponds of the warmer parts of the
world. Pistia has a high growth-rate and causes
harm to water resources and agriculture in the
same manner as other major aquatic weeds. It
might be profitably used as a substrate for biogas
production.

METHODS
Sampling of P/st/a
Fresh Pistia (whole) plants were collected from
the Botanical Garden, University of Graz,
Austria. The plants were washed with water to
remove attached sediments, and then finely
ground in an 'Ultra Turrax' homogenizer (Janke &
Knkel, Staufen, Germany).
Source of inoculum
Digested slurry from a biogas plant (Harrer
Biogas Plant, Graz) running with cow manure at
35C for several years, was used as an inoculum
for the present study.
Chemical analysis of P/st/a and inoculum
Total solids (TS), volatile solids (VS), nitrogen (N)
and pH were determined as the standard methods
(Rand et al., 1985). Ammonium nitrogen and
chemical oxygen demand (COD) analyses were
carried out according to the German Standard
Methods of the examination of water, wastewater
and sludge (Frachgruppe Wasserchemie, 1985).

211
Bioresource Technology 0960-8524/91/S03.50 1991 Elsevier Science Publishers Ltd, England. Primed in
Great Britain

S.A. Abbasi, P. C. Nipaney, M. B. Panholzer

212

Volatile fatty acid (VFA) and methane analysis

Volatile fatty acids and methane were estimated
using a Hewlett Packard gas-liquid chromatograph.

Anaerobic digestion of Pistia

All-glass air-tight digesters of 3000 ml capacity
with gas sampling and measuring units were
employed. Each digester set consisted of a 3000ml round-bottom flask as digester, calibrated gas
collecting tube and a bottle to receive displaced
water. The digesters had two outlet connections;
one for gas and one to collect the slurry for
analysis. The gas-collecting tube was filled with
acidified water (pH N 2) to prevent absorption of
carbon dioxide and hydrogen sulfide gases from
the biogas.
The digesters were charged with Pistia or
Pistia-inoculum mixtures. The controls were with
inocula alone.

RESULTS AND DISCUSSION

Some characteristics of the feed are presented in
Table 1.

Biogas yield
The biogas yield per kg (fresh weight) of the feeds
over a 30-day retention time is presented in Table
2. The methane content of the biogas from
digesters A and C (inoculum/inoculated Pistia)
was in the range 52-54% during the first 2 days of
the digestion and remained in the range 61-68%
for the remaining period. The biogas from
digester B (Pistia without inoculum) consisted
only of carbon dioxide.
As might be expected the presence of inocula
has a very pronounced positive effect on the biogas production from Pistia.
The pattern of biogas yield -- HRT curves (not
shown here) for Pistia was similar to the patterns
Table 1. Some characteristics of the feed (wet weight basis)

Parameter

lnocula

Total solids (TS%)

Volatile solids (VS%)
Chemical oxygen demand
(g O2 kg -1)
Ammonium nitrogen
( g N k g -~)
Kjeldahl nitrogen
( g N k g -~)

6.9
5.4
67

Pistia
4.6
3'9
36

Mixture
5.6
4-4
62

2.2

0.05

1"9

3-7

0-6

3'2

observed with Ceratopteris but different from the

ones observed with Salvinia, Azolla, Hydrilla,
Nymphaea, Cyperus, Scirpus and Utricularia
(Abbasi & Nipaney, 1990). The nature of VS in
Pistia is such that the bulk of degradation takes
place during the initial phase of the digestion, suggesting that the digesters with Pistia should preferably be run at an HRT close to 10 days for
optimal energy yield.

pH
The pH of the digesters running on inocula or
Pistia-inoculum mixtures was in the mildly
alkaline range 7.4-8.0 whereas the digesters with
"Pistia alone remained significantly acidic; the pH
never rising above 5"3 and often remaining lower
than 5 (Tables 3 and 4).

VFA Composition
The nature and composition of volatile fatty acids
formed during the course of the digestions are
shown in Tables 3 and 4. It may be seen that in the
case of Pistia, the acetic acid concentrations shoot
up to 517 ppm by the 4th day of the digestion and
remain in the range 563-978 ppm thereafter,
indicating that while the plant is contaminated
with fermentative bacteria, methanogenic bacteria
are absent from Pistia plants. In contrast inoculated Pistia has a low concentration ( < 70 ppm) of
residual acetic acid from the 8th day onwards.

Table 2. Cumulative biogas production from inocula, Pistia

and inoculated Pistia

Number
of days
2
4
6
8
10
12
14
16
18
20
22
24
26
28
30

Biogas production, 1 kg- i (fresh weighO

Digester A"

Digester B b

Digester C"

1'10+0'00
1"95+0"05
2"80 + 0"00
3.55+0"05
4'15+0"05
4"70+0"10
5.35+0"05
5'75+0"05
6'25+0"05
6"65+0'05
7-05+0"05
7-35_+0"05
7"80+0"00
8.00+0.00
8'40+0"00

1'70+0'00
2"00+0"00
2"00 + 0.00
2"15+0"15
2.45+0.15
2"45+0"15
2'80+0"20
2'80+0"20
3'15+0"15
3"15+0"15
3-15+0'15
3-30_+0"00
3'45+0"15
3.45 +0"15
3"45-+0"15

7-70+0"10
14"7+0"30
17"95 + 0.65
19"25+0"65
20"20+0"60
20"65+0"65
20-80+0"70
21"25+1.15
21"35+1"25
21'50+1"20
21'90_+1"20
22"15+1.25
22"40+1"10
22"45 + 1"05
22"80+0.70

All values represent the averages of two digesters.

"Digester A: Inoculum = 2000 g.
bDigester B: Pistia = 300 g; water = 1500 ml.
'Digester C: Pistia = 300 g; inoculum = 1500 g. Temperature:
35C.

Biogasfrom Pistia

The digesters charged with Pistia alone have significant concentrations of higher fatty acids
including propionic, butyric, isobutyric, valeric,
and isovaleric acids. In the case of inoculated
Pistia propionic acid, only, was detected during
the first 4 days of the digestion. Inoculated Pistia
has even lower residual acetic acid levels than the
inoculum alone.
Effect of inocula on a 'soured' digestion
Inoculum (1000 ml) was added to the 'soured'
digesters (pH 5.0) which had already run on Pistia
as feed for 30 days. The gas production on the
30th day, and the cumulative gas production -consisting solely of CO: -- were 0 and 3.5 liters
respectively before the addition of inoculum. The
concentration of acetic, propionic, butyric, isobutyric, valeric and isovaleric acids at that time

213

were appreciable. Soon after the addition of the

inoculum this situation changed dramatically
(Table 5). Within 24 h the concentration of acetic
acid, propionic, isobutyric, and butyric acids
dropped considerably. By the second day the
acetic acid concentration had fallen to 323 ppm
and that of propionic acid to 11 ppm. The other
acids had been consumed to below detectable
levels. The gas production also picked up and
methane appeared in significant levels (60%) on
the first day, building up to 63% by the 7th day.
The cumulative gas production per kg VS of Pistia
in 25 days was 459 liters compared to 573 liters
obtained when Pistia was inoculated at the outset.

Table 5. pH and volatile fatty acids in an inoculated 'soured'

digester
Initial VFA concentrations:
lnocula
221 mg liter- i

Table 3. pH and volatile fatty acids in digesters with

inoculum or Pistia + inoculum

Number

pH

Pistia

978
109
113
273
47
31

Volatile fatty acids (rag liter- i)

of days
lnoculum
Acetic
2
4
6
8
10
15
20
25
30

7-5-7"7
7.5-7.7
7'5-7.6
7.6-7.7
7.5-7.6
7"5-7-7
7"5-8"0
7-6-7-7
7-4-7.5

154_+3
155_+5
131_+1
110+-2
130_+9
91_+5
92+-1
83_+2
102_+6

Pistia + inoculum
Acetic

acetic acid (A)

propionic acid (P)

3 mg liter J

acetic acid (A)

propionic acid (P)
isobutyric acid (iB)
butyric acid (B)
isovaleric acid
valeric acid

mg liter I
mg liter- ~
mg liter- ~
mg liter- L
mg liter- ~
mg liter- ~

Propionic

531_+35
209_+6
90_+8
65+-11
60_+1
44_+5
43+-2
39_+2
63_+3

75+-20
ND ~
ND
ND
ND
ND
ND
ND
ND

Number
of days a

lnoculum
pH

7.8
7.7
7.4
7.4
7.3

71+-3
50+-0
59_+0
39_+3
33_+2

1
2
4
10
25

Inoculated Pistia
pH

7'5 627_+3
7.5 323_+5
7.5
55_+9
7.4
27+-2
7.4
34_+2

51+-2
11_+1
ND
ND
ND

iB

9
ND
ND
ND
ND

ND
ND
ND
ND
ND

All values represent the average of two digesters.

"After inoculation.
ND = Not detectable.

All values represent the averages of two digesters.

aND = Not detectable.

Table 4. pH and volatile fatty acids in digesters with P&tia alone

Number
of days

pH

Volatile fatty acids (mg liter- 1)

Acetic

2
4
6
8
10
12
14
16
20
25
30

4-5-5"3
4.7-5"1
4-9-5"0
4'9-5'0
4'9-5"0
5-9-5"0
4"8-5"0
4"7-4"8
4"8-4"9
4'9-5"0
4'9-5'0

289 + 33
517 + 115
5 6 3 + 100
6 3 0 + 117
734 + 115
782_+ 125
833 + 119
851+_117
939 + 122
904-+94
978_+ 108

Propionic

lsobutyric

3+0
72 ___38
101 + 4
109+7
120 + 8
117+ 11
122 -+ 10
127+-8
142 _+9
94-+ 17
109+-4

ND
5+ 2
8+2
10+2
19 + 5
23+6
34 -+ 10
45_+12
74 +- 14
7 3 + 12
113_+4

All averages represent the averages of two digesters.

Butyric
150 + 34
245 + 1
249+0
267+ 8
282 + 6
282+2
281 _+ 12
297_+8
297 + 12
222_+ 13
273_+37

lsobutyric

Valeric

ND
ND
7+7
11 + 6
28 + 1
48+2
61 + 8
42+7
81 + 12
54_+26
47+7

ND
ND
ND
ND
14 + 5
19-+ 1
24_+ 9
31+-14
35 -+ 18
25+9
31 _+7

214

S. A. Abbasi, P. C. Nipaney, M. B. Panholzer

Pistia could, then, be used as a substrate for

biogas production provided that it was fed to a
working continuous-flow digester or that an
inoculum was used for a batch digester. Biogas
production could be an incentive to harvest the
weed, so reducing pollution of water resources.

ACKNOWLEDGEMENT

The work was financially supported in part by

'Bundesministerium for Auswatige Angelegenheiten', Vienna, Austria and in part by Council of
Scientific and Industrial Research, New Delhi; the
authors gratefully acknowledge this support.

REFERENCES
Abbasi, S. A. & Nipaney, E C. (1990). Bioenergy potential of
eight common aquatic weeds. Biol. Wastes, 34, 359-65.
Attionu, R. H. (1976). Some effects of water lettuce on its
habitat. Hydrobiologia, 50, 245-54.
Bua-Ngam, T. & Mercado, B. L. (1975). The life cycle of
water lettuce (Pistia stratiotes L.). Philippine Weed Science
Bulletin, 2, 11-15.
Fachgruppe Wasserchemie (1985). Deutsche Einheitsverfahren zur Wasseruntersuchung. Verlag Chemie,
Weinheim, Germany.
Rand, M. C., Greenberg, A. R. & Taras, M. J. (1985). Standard Methods for the Examination of Water and Wastewater,
16th edn. APHA, Washington DC.
Rao, E N. & Reddy, A. S. (1984). Studies on the population
biology of water lettuce: Pistia stratiotes L. Hydrobiologia,
l l 9 , 15-19.
Tadulingam, E L. S. & Venkatanarayana, G. (1985). A Handbook of Some South Indian Weeds. Periodical Expert
Book Agency, New Delhi, pp. 365.