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Bioresource Technology 4 1 (1992)259-264

Methanogenic Fermentation of Cassava Peel Using a

Pilot Plug Flow Digester
N. Cuzin,"" J. L. Farinet,bC. Segretain" & M. Labat
"ORSTOM,Laboratoire de Microbiologie,BP 181,Brazzaville, Congo
*LRAT/CIRAD,BP 5045,34032 Montpellier cédex, France
(Received 10 October 1991; revised version received 9 November 1991; accepted 12 November 1991)

Abstract cassava peel that could be used to produce biogas

and to generate part of the energy needed for the
During the methanogenic fermentation of cassava mechanical processing of cassava (about 1200
peel, its composition (high starch content, high kWh required to dry 1 ton of cassava meal). How-
carbon :nitrogen ratio, presence of cyanogenic ever, the high starch content (85% of dry matter),
glucosides) usually results in excess acid produc- the high) C:N ratio (76) and the presence of
tion, nitrogen deficiency and the release of cyanide, cyanogenic glucosides in cassava peel can induce
which is highly toxic to methanogenic bacteria. excess acid production, nitrogen deficiency and j!,

The utilization of a plug flow digester ('Trans- the release of cyanide, which is highly toxic to
paille') solved the problem of acidification through methanogenic bacteria (Eikmanns 9r Thauer, *.

localization of the acidogenic phase in the first half 1984; Smith et al., 1985).An inhibitory effect of
of the fermenter and the problem of nitrogen high acid production and N deficiency on cassava
deficiency because a permanent liquid phase waste methanogenesis in fermenters was reported
allowed nitrogen accumulation. No perturbation by Wurster (1985) and Segretain and Bories
due to cyanide (5-6 mgIl) was observed in the fer- (1987).
menter. We studied the metb.ane production potential
A fermentation yield of 0.661..m3biogaslkg vola- of raw cassava peels i n a 128-liter plug flow
tile solids (VS) was obtar'yed with a loading rate of digester. The aim of the study was to define an
3.6 kg VS/in3 day. Energy-saving calculations appropriate fermentation technology so that bio-
showed that a fermenter of 88m3 is needed to gas can be used instead of' electrical energy to dry
produce the methane necessary for drying one ton cassava meal.
of cassava meal.

Key words: Cassava peel, anaerobic digestion, METHODS

biogas, fermentation yield. m
m
5 3
'Transpaille' digester =F--

The digester used a continuous process patented

INTRODUCTION by IRAT/CIRAD (France). It is based on the
transfer of a heterogeneous substrate immersed in
The processing of cassava roots to produce cas- water in a horizontal cylindrical tank (Fari.net et I
p

sava meal (fufu) in the Congo includes peeling, al., 1987). In the fermenter used for the experi-
retting by soaking roots in water for three days ment the mobile solid phase undergoes a plug- 2 y
flow pattern movement (Fadlalla, 1989) and is 2 s
.I

and drying. During this process, large quantities of

solid wastes (cassava peel) are produced. In the fermented in a nonrenewed liquid phase estab- -f
Congo, the annual production of 700 O00 tons of lished once at the beginning of the process ZQ
cassava roots leads to 175 O00 tons of unutilized (Fadlalla, 1989). s31-
The fermenter (Fig. 1)includes three parts: gT
*To whom correspondence should be addressed at:
ORSTOM, Université de Provence, 3 Place Victor Hugo, - A feeding box in which the substrate is
szF'si2
.3
13331 Marseille Cédex 3, France. loaded. The feeding box is equipped with a IB 0

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259 4-

Bioresource Technology 0960-8524/92/$05.00 O 1992 Elsevier Science Publishers Ltd, England. Printed in %z
Great Britain
 260 N. Cuzin, J. L. Farinet, C.Segretain, M. Labat

\,,,,i,?&
Feeding box(30litres) Fermentation tank (128 litres) Effluent box (220litres)

Mosquito s c r e e n
Wooden box
' G2

Mobile plate Stroke of t h e plate älades Heating coil

during loading
Fig. 1. 'Transpaille' fermenter - longitudinal cross-section. G1, G2, G3, Gas sampling points; C1, C2, C3, liquid sampling
points; 1,position for loading; 2, position during fermentation. See text for detailed explanation.

mobile plate connected to a jack which studied for 600 days. Gas production was
pushes the solid substrate and ensures the measured daily, pH and temperature were
closure of the fermentation tank after load- measured twice a week, chemical oxygen demand
ing, Blades attached to the jack axis help to (COD), a l k a h t y and volatile fatty acid (VFA) I

move forwards the solid phase in the fer- were measured twice a month, and gas composi-
% mentation tank during loading. When the tion was measured once a month.
,fermentation tank is full of substrate, each
new loading induces the evacuation of Substrate
digested substrate in the effluent box. Cassava peels were collected from an industrial
- A fermentation tank. Three gas sampling cassava farm, located in an equatorial savannah
points (Gl, G2, G3) and three liquid sampl- region (Mantsoumba, Congo). Fresh peels were
ing points (Cl, C2, C3) are installed along stored at 4°C until use.
the tank. Temperature (35-39°C) was main-
tained by circulating hot water in a coil sur- Analytical methods
rounding the tank. A 4% slope-of the tank The total; solid (TS) content of peels was deter-
allowed biogas collection in an exhaust miaed by drying at 105°Cfor 24 h and the volatile
equipped with a flowmeter. solid (VS) content by calcination at 500°C. I

- An effluent box open to the air. *' Nitrogen was measured using a Kjeldahl method.
Chemical oxygen demand (COD),alkalinity to pH
The digester was completely filled with water. 4-2 and potassium and phosphorus contents were
Straw (87-90% TS (total solid); 90-9570 VS estimated using standard methods (American
t, (volatile solid)) at twice the feeding box capacity Public Health Association, 1985). Cyanide was
was introduced into the fermenter to form a plug. determined as described by Cooke (1979).Biogas
Bovine rumen content ( 10% of the useful volume) composition was monitored by gas chromato-
was loaded for inoculation. No additional inocu- graphy on a DELSI 30-E chromatograph (Pora-
lum was needed. pak Q column, 2 m, in, 80°C, carrier gas N, 25
During the four first months of incubation the ml/min).The same apparatus was used for volatile
fermenter was loaded every three days with a fatty acid ( V A )analysis (Haye Sep Deb column,
mixture (2.2kg VS/m3 day) of raw cassava peel - 1 my in, 18O'C, carrier gas N, 2 5 &/min).
90% TS - and straw - 10% TS. The retention
time of the solid phase was about 45 days. Tech- Enumeration of digester microflora
nical problems delayed the reaching of the steady Fermentation liquid (100 ml) was collected
state (deficiency of substrate, gas leak, etc.). anaerobically from sample points of the digester.
At steady state, the fermenter was loaded Media and inoculation techniques were as des-
manually once every two .days with cassava peel cribed by Hungate (1969) and Balch et al. (1979).
only. The criterion for determining that steady Amylolytic, fermentative, sulfate-reducing and
state had been reached was the stability of gas methanogenic bacteria were counted by the most-
production. Cassava peel fermentation was probable-number (MPN) technique with five
 “l I

8‘ J

Methanogenic fermeiztatioiz of cassava peel 26 1

I

tubes per dilution. The organisms were grown in

the medium of Balch et al. (1979). Starch ( 5 g/l) 7.0
was used as substrate for amylolytic bacteria,
glucose (5 g/l) for fermentative bacteria and 5 6.0
sodium acetate ( 5 g/l)+methanol ( 0 5 g/l) for
acetoclastic bacteria. For the growth of hydro- 5-0
genotrophic bacteria, the N,/CO, (4:1) atmos-
4.0
1 ‘Addition of NapCO,
. . , . .
2.2 kg VSlm3 day
phere was replaced by H2/C02 (4:l).
I . 1 I

O 20 40 60 80 100 120
Sulfate-reducing bacteria were grown in the Days
medium of Widdel (1980). The substrate was (4
either sodium lactate (2-5g/l) or sodium acetate ( 2
g/l). Incubations were performed at 37°C. The 8.0 ~

rhodanese activity was determined as described

by Singleton and Smith (1988).

RESULTS
3.6 kg VS/m3 day
5.0
’ 295 300 305 310 315 320 325
The composition of cassava peel was 25-35% Days
TS, VS was 90-97% of TS, starch 65-85% of TS,
(b)
carbon 33-46% of TS, nitrogen 0.6% of TS, phos-
phorus 0.7% of TS, potassium 0.9% of TS and
cyanide up to 400 mg/kg fresh matter.
Three levels of loading were studied during the
methanogenic fermentation: the start-up loading
rate averaging 2-2 kg VS/m3 day with 10% straw
and 90% cassava peel, and two steady states of 3.6
and 4.2 kg VS/m3 day with 100% cassava peel. 4.2kg VS / m 3 day
The average biogas production at 37°C was 1.40
515 520 525 530 535 540 545
m3/m3day with a load of 2.2 kg VS/m3 day, 2-38 Days
with 3.6, and 2.57 with 4*2..Themean yield was (cl
I

0.629 m3 biogaslkg VS (0.661 from 3.6 kg VS/m3

Fig. 2. pH measured at the liquid sampling points along the
day), which amounts to 0.217 m?/kg fresh cassava tank ( 0 C1, 0 C2, A C3) during (a) the start-up of the fer-
peel (or 0.065 m3/kg cassava roots, with a peeling mentation and at stabilized loading rategof (b)3-6 and (c)4-2
yield of 3Oo/O).The mean methane content of the kg VS/m’ day.
biogas was 57%, but the level varied from one
sampling point to another. The highest percentage .”
was recorded at point G3 (75%). The COD in the liquid phase was 5 g/1 in the
During the start-up of the fermentation, the pH fsding box and 2-5 g/l in the pit at steady states.
dropped to 5 in the-firstpart of the reactor (feed- The total alkalinity varied around 3.8 g CaCO,/l,
ing box, C1 and C2) and was corrected by adding when no VFA was measured.
Na2C03. The pH was around 6.5 at sampling Some acetate (0.3 g/l) and propionate (2 g/l)
point C3 and in the pit (Fig. 2(a)). After 10 accumulated in the first part of the reactor when
months of operation, when the loading rate was the loading rate was 3-6 kg VS/m3 day. After one
3-6 kg VS/m3 day, pH increased to about 7.0-7.6 month, these VFA concentrations decreased and
in all parts of the fermenter (Fig. 2(b)).After 17 VFA were then no longer detectable.
months when the loading rate was 4.2 kg VS/m3 During the longest period of stable loading rate
day, pH remained around 7.1-7.3 in the first part obtained in this experiment (3.6 kg VS/m3 day
of the fermenter (feeding box, C1, C2) and between 294 and 500 days) the NH,f-N increased
reached 8 in the second part (C3 and pit) without slightly (0-05-0-2 g/l), while total-N markedly
any further pH correction (Fig. 2(c)).Dynamics of increased (0.8 to 2 g/l), without any addition of
pH measured at the feeding box and C1, and at nitrogen (Fig. 3(a)and (b)).
C3 and the pit were very similar. Therefore only The results of the MPN counts performed at
values for C 1,C2 and C3 are presented in Fig. 2. the three liquid sampling points (Cl, C2 and C3)
 262 N. Cuzin,J. L. Farinet, C.Segretain, M. Labat

at stabilized loading rates (3.6 and 4.2 kg VS/m3 scent of Methanobacterium and rods combined
day) showed that bacterial density increased with end to end in long filaments as in Methanusaeta.
the loading rate (Table 1).In the first half of the The methanogenic bacteria isolated were related
fermenter (C1 and C 2 ) , glucose degraders pre- to Methanubacterium and Methanosarcina and
dominated over the methanogenic bacteria, which were highly sensitive to cyanide in pure culture
were the main group in the second half (C3). As ( < 1 mg/l). The cyanide levels in the digester
compared with other bacterial groups, sulfate- reached 5-6 mg/l, without perturbing biogas pro-
reducing populations were low and their number duction. We isolated a sulfate-reducing bacterium
increased with the loading rate, mostly in the first belonging to the genus Desulfovibriu that toler-
part of the fermenter. The methanogenic micro- ated up to 25 mg CN/1 and exhibited a rhodanese
flora contained a variety of organisms including activity known to detoxify the cyanide-producing
large cocci and aggregates resembling those of thiocyanate.
Methanusarcina, thin filaments similar to those in Feeding was stopped for several weeks to
Methanuspirillum, a large number of rods remini- improve the gas exhaust of the digester. At restart,
a too-high loading rate resulted in acidification at
the entrance of the fermenter. This allowed us to
h

.
p1
P
200-

150
P study how the digester responded to an accidental
acidification. Chemical oxygen demand increased
to 30 g/1 in the feeding box, whereas pH
100 decreased to Sk and 10 g/1 acetate accumulated.
o,
v
E 50 This acidification increased along the cylindrical
tank between points C1 and C2. The bacterial
O
300 320 340 360 380 400 420 440 460 480 5 count showed 10 times less fermentative bacteria
Days and 100 times less methanogenic bacteria than at
(4 the previous steady state. Biogas production was
-
Q,
L
2500)
*
reduced to 20% of the normal production at the
same loading rate before acidification. To correct
s 2000- this failure, loading was stopped and inoculum
? 1500- (rumen content) was added in the feeding box.
+A
O
;1000- After several days, the conditions (COD, pH, etc.)
E became optimal for methanogenic fermentation;
500-
only a marked increase in NHZ-N and total-N
ir
O
300 320 340 360 380 400 420 440 460 480 5
was recorded. \r-
Days
(b)
DISCUSSION
Fig. 3. Nitrogen levels, (a) NHZ-N and (b) total-N in the
liquid phase ( O feeding box, C1, 0 C2, A C3, A effluent
box) during the anaerobic fermentation of cassava peelat the The COD and pH values of the liquid phase
stable loading rate (3.6 kg VS/m3day). showed that most of the organic matter seemed to

Table 1. Enumeration of bacteria in fermenter juice from three liquid sampling points along the reactor (Cl, C2, C3)
~

Metabolic group Substrate Loading rate:

statistical MPN evaluation of organisms (X 107/ml)
3.6 kg VS/m-3day 4.2 kg VS/m3day
CI C2 c3 CI c2 c3
Amyiolytics Starch nd nd nd 80 130 3.5
Acidogens Glucose 5.0 35 0.80 50 50 3.5
Sulfate-reducers Lactate 0.35 0.35 0.80 2.5 2.0 0.80
Acetate 035 0.25 0.50 9.5 0.70 0.45
Methanogens H2ICO2 8.0 25 35 25 35 250
Acetate 0.13 0.17 0.25 3.5 0.40 25
nd, Not determined.
a‘ J

Metlianogenic fermentation of cassava peel 263

be degraded in the first part of the fermenter, tons) to produce methane providing 100% of the
whereas production of biogas occurred in the energy necessary for drying one ton of cassava
second part. Increasing CH4 concentrations from meal. The volume of the ‘Transpaille’ digester
G1 to G3 sampling points agree with this hypo- needed to produce this 121 m3 CH, would be
thesis. Bacterial counts showed that acidification 88 m3.
occurred early on in the first part of the fermenter, The production of biogas from anaerobic
where amylolytic and fermentative bacteria were digestion of cassava peel in a ‘Transpaille’digester
predominant. Methane production was higher in provides an important energetic potential, which
the second part of the fermenter, where methano- should be of value in improving the economics of
gens were predominant. cassava processing.
The piston flow of the solid fraction (Fadlalla,
1989) may favor the progressive elimination of
intermediate acid compounds during substrate
transit and prevent drastic acidification. The ACKNOWLEDGEMENTS
‘Transpaille’ system showed a satisfactory
capacity to cope with acidification. Nitrogen The authors are grateful to Edouard Miambi for
analyses showed a progressive nitrogen enrich- stimulating discussions, Guy Eboungabeka for
ment in the liquid phase. The cassava peel was technical assistance and Hervé Macarie and
loaded undiluted and the solid effluents were dis- Pierre Roger for reviewing the paper.
charged after being drained; therefore the reactor
liquid volume did not vary significantly. In the
‘Transpaille’ digester, a progressive nitrogen REFERENCES
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