The 1st IWWG-ARB Symposium, 18-21 March 2013, Hokkaido University, Japan

Failure Analysis on Co-digestion of Agricultural Residues and Food

Waste: Focusing on Heavy Metal, Trace Element, Methanogenic
Activity and Microbial Acclimation

Y. J. Lee and J.Y. Kim*

Department of Civil and Environmental Engineering, College of Engineering,

Seoul National University, Seoul, Republic of Korea
*Corresponding author: jaeykim@snu.ac.kr

1. INTRODUCTION

Anaerobic digestion producing methane-rich biogas has been used as a source of

renewable energy. The Korea government decided to establish bioenergy villages
where organic wastes generated in the villages are used as energy sources. In the
agro-type village, agricultural residues from plastic greenhouse and food waste have
been considered to be important substrates for biogas production.
Considering the fact that cultivation area of plastic greenhouse is expending every
year (Statistics Korea, 2009) it is expected that the amount of agricultural residue
production will increase continuously. However, limited data is available on
agricultural residue digestion produced in plastic greenhouses. Food waste is another
major organic waste. It is considered as desirable substrate for anaerobic digestion
due to its high biochemical methane potential of 390-490 mL-CH4/g-VS (Heo et al.,
2004; Moon et al., 2011). However, problem was often observed in the food waste
digestion due to accumulation of volatile fatty acids (VFAs) and free ammonia that
leads to deterioration of performance and possibly process failure (Heo et al., 2004).
Nowadays, anaerobic co-digestion has attracted more attentions because it can
offer benefits such as dilution of inhibitory compounds, improved nutrient balance,
increased biogas production and high cost efficiency. These practices suggest that
agricultural residues and food waste co-digestion could overcome process limitations
and low economic feasibility found in anaerobic digestion when they are digested
alone.
The objective of this study was to evaluate the feasibility of the co-digestion of
agricultural residues and food waste under mesophilic conditions. Additionally, in
order to identify the factors of unstable digestion, methanogenic activity, microbial
adaptation, heavy metal and trace elements were investigated.

2. MATERIALS AND METHOD

2.1. Feedstock

Stems and leaves of tomato and cucumber after harvesting were obtained from the
agricultural plastic greenhouses in South Korea. Artificial standard food waste was
prepared with 10 types of food by referring to the components of food waste
following the standard of Korea Environmental Industry & Technology Institute.
They were ground using electrical kitchen blender and thereafter stored frozen at -10
oC prior to thawing and used in experiments.
 The 1st IWWG-ARB Symposium, 18-21 March 2013, Hokkaido University, Japan

2.2. Operation of continuously stirred tank reactor (CSTR)

Semi-continuous digestion of food waste alone and co-digestion of were carried in

4 L CSTR of 2 L working capacity at 351oC. The digesters were fed once per day
and the reactor was constantly mixed using mechanical stirrers at 300 rpm to ensure
the homogeneity of the liquid phase. Table 1 describes the composition of the
feedstock. The ratios were chosen based on organic waste production amount from
sample bioenergy village.

Table 1 - Composition of feedstocks for anaerobic digestion reactors

Reactor Composition of feedstock (on VS basis)
CSTR 1 49% Cucumber residue / 51% Food waste
CSTR 2 52% Tomato residue / 48% Food waste
CSTR 3 100% Food waste
CSTR 4 100% Cucumber residue
CSTR 5 100% Tomato residue

Five series of experiments were conducted to evaluate the feasibility of anaerobic

co-digestion of food waste and agricultural residues and to identify the factors
responsible for unstable digestion. In experiment 1, food waste digestion and co-
digestion were operated at 1g-VS/L-day of organic loading rate (OLR) and 30 days of
hydraulic retention time (HRT). In experiment 2 for failure analysis of experiment 1,
methanogenic activity assays were carried out to check the activation of
methanogenesis in reactor. For this experiment, batch test were followed by Owen et
al. (1979) and the modified Gompertz equation was used to evaluate the rate of
methane production and lag time from experimentally observed methane production
(Lay et al., 1997). In experiment 3, to acclimate the microbes to operation condition,
OLR was raised stepwise from 0.1 to 1g-VS/L-day (Table 2). Once digester was
stabilized at OLR of 1g-VS/L-day, it was operated for long term. In experiment 4, for
the digesters still unstable after acclimation, possibility of heavy metal toxicity was
investigated. The heavy metal concentrations (Cu, Zn, Ni) in the effluent were
analyzed. Finally, the effect of trace elements on anaerobic digestion was investigated.
The medium of trace elements was followed by the description of Shelton and Tiedje
(1984) and it was injected once during the experiment.

Table 2 - Reactor operating conditions during microbial acclimation stage

OLR (g-VS/L-day) HRT (day)
Phase 1 0.1 300
Phase 2 0.25 120
Phase 3 0.5 60
Phase 4 0.75 40
Phase 5 1 30

2.3. Analytical methods

 The 1st IWWG-ARB Symposium, 18-21 March 2013, Hokkaido University, Japan

Total Solid (TS), Volatile Solid (VS) were analyzed according to Standard
Methods (APHA, 2005). The methane concentration in the biogas was analyzed using
gas chromatography (GC) (Younglin, Korea), equipped with a thermal conductivity
detector (TCD) operated at 120C, with injector and oven temperatures of 120 and
35C, respectively. VFAs (acetic acid, propionic acid, iso-butyric acid, butyric acid,
iso-valeric acid, valeric acid, hexanoic acid) were analyzed using GC (Younglin,
Korea) equipped with flame ionization detector (FID) and HP-INNOWAX (Aglient
Technologies, USA). The injection and FID temperature were 240 and 250oC,
respectively. The initial oven temperature was 120oC for 2 min, with a 10oC/min ramp
to 190oC. Trace metal analysis was performed using an ion coupled plasma-atomic
emission spectrometer (ICP-AES) (OPTIMA 4300DV, Perkin-Elmer, USA). Heavy
metals were assayed using Flame Atomic Absorption Spectrometer (AA-7000,
Shimadzu, Japan).

3. RESULT AND DISCUSSION

3.1. Anaerobic digestion of food waste and co-digestion

The semi-continuous digestion of food waste and co-digestion were operated and
the specific methane productions for all digesters are shown in Figure 1. Compared to
previous studies, the operation condition was set at low OLR of 1 g-VS/L-day in
order to avoid the overload (Ganaseelan, 1997; Heo et al., 2004; Moon et al., 2011).
Even though the conditions were not extreme, within 10 days of operation,
methane production stopped and pH had dropped below 6.5 for CSTR 1, 3 and steady
state was not reached. For CSTR 2, 50-100 mL/g-VS of methane was produced but
methane contents and pH were below 40% (by vol.), 7, respectively causing unstable
performance. At the end of the experiment, total VFA concentration of CSTR 1, 2, 3
were 9,000, 5,000, 8,300 mg/L, respectively. Compared to previous study of stable
standard food waste digestion (Moon et al., 2011), high propionic acid concentration
of 2,000-2,300 mg/L was observed. Propionic acid concentration above 900 mg/L can
cause significant inhibitory effect on methanogens (Wang et al., 2009). Therefore, in
this study, the reactor performances were failed due to propionic acid accumulation.

400

CSTR 1
Methane production (mL-CH4/g-VS)

CSTR 2
CSTR 3
300

200

100

0
0 5 10 15 20 25

Time (day)

Fig 1. Profiles of methane production of anaerobic digestion at OLR of 1 g-VS/L-day

and HRT of 30 day
 The 1st IWWG-ARB Symposium, 18-21 March 2013, Hokkaido University, Japan

3.2 Methanogenic activity assay

Methanogenic activities of the sludge from CSTR 1, 2, 3 were measured. The

initial lag phase in the production of methane was observed for all tests and its value
was in range of 5-6 days. Methanogens in all reactors could convert each substrate to
methane and methane production rate of CSTR 1, 2, 3 were 23.3, 12.3, 23.6 mL-
CH4/g-VS day, respectively. After 50 days of investigation, cumulative methane
productions of CSTR 1, 2, 3 were 321, 262, 374 mL-CH4/g-VS showing that methane
production of food waste was the highest and mixture of food waste and tomato
residue was the lowest.

3.3 Microbial acclimation

Changes in OLR and methane production of CSTR 2 during the acclimation period
are shown in Fig 2. After raising OLR to 1 g-VS/L-day, stable performance was
observed more than 70 days. In this condition, methane production ranged from 270
to 420 mL-CH4/g-VS and total VFA concentration was below 2,000 mg/L. Also,
methane contents and VS removal efficiency ranged 53-56% (by vol.), 70-75% (by
wt.), respectively.

Fig 2. Methane production of CSTR 2 during microbial acclimation stage

On the other hand, unstable digestion process leading low methane production or
VFA accumulation was observed in CSTR 1, 3, 4, 5 during microbial acclimation
process.

3.4 Heavy metal toxicity

Among microbial acclimation failed reactors, CSTR 1, 4 showed high

concentration of copper in effluent (Table 3). In these reactors, cucumber residue was
fed as feedstock and it was observed that raw sample of cucumber residue contained
relatively high copper concentration (349 mg/kg) compared to tomato residue (14
mg/kg) and food waste (11 mg/kg).
Table 3 - Concentrations of heavy metal (Cu, Zn, Ni) in effluent (mg/L)
 The 1st IWWG-ARB Symposium, 18-21 March 2013, Hokkaido University, Japan

CSTR 1 CSTR 2 CSTR 3 CSTR 4 CSTR 5

Cu 1.3 nd nd 2.7 0.8
Zn 3.6 1.2 0.7 1.1 1.0
Ni 0.1 nd nd nd 0.1
nd: not detected (detection limit : Cu > 0.2 mg/L, Ni > 0.05 mg/L)

The significant difference in inhibiting heavy metal concentration can be attributed

to the differences in sludge, environmental conditions and acclimation period (Lin,
1993; Shin et al., 1999; Ahring et al., 1983). Heavy metal concentrations that caused
50% inhibition of VFA degradation during glucose digestion were 0.9, 3.5, 440 mg/L
for Cu, Zn, Ni, respectively and the results indicated that toxicity decreased in the
order of Cu > Zn > Ni (Lin, 1993). Considering the previous study, it is assumed that
high copper concentration in cucumber residue might have caused inhibition in
digestion performance.

3.5 Effects of trace metal supplement

After supplementing trace metal medium, stable performance was observed in

CSTR 3 for more than 40 days. At OLR of 1 g-VS/L-day, methane production ranged
480-600 mL-CH4/g-VS (Fig. 3). Methane contents and VS removal efficiency ranged
54-57% (by vol.), 83-85% (by wt.), respectively. Inhibition due to VFA accumulation
was not observed throughout the experiment.

Fig 3. Methane production of CSTR 3 after trace element addition

Similar results were obtained by Zhang et al. (2011) which demonstrated that trace
metals (Co, Mo, Ni, Fe) played an essential role for stable food waste digestion. This
implies that food waste was deficient in trace elements required for robust and stable
anaerobic digestion.
 The 1st IWWG-ARB Symposium, 18-21 March 2013, Hokkaido University, Japan

4. CONCLUSION

The co-digestion of tomato residue and food waste was stable in operation at OLR
of 1 g-VS/L-day and HRT of 30 days. In this condition, methane yields ranged 270-
420 mL-CH4/g-VS and VS removal efficiency showed 70-75% (by wt.). The results
of failure analysis suggested that heavy metal, trace metal and microbial acclimation
could affect anaerobic digestion performance when treating agricultural residues and
food waste. It is expected that the results will provide important information to
achieve stable and efficient anaerobic digestion process in bioenergy villages.

ACKNOWLEDGEMENT

This study was supported by SNU SIR Group of the BK21 Research Program
funded by Ministry of Education, Science and Technology.

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