Bioresource Technology 126 (2012) 425–430

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Effect of inoculum to substrate ratio on the hydrolysis and acidification of

food waste in leach bed reactor
Su Yun Xu, Obuli P. Karthikeyan, Ammaiyappan Selvam, Jonathan W.C. Wong ⇑
Sino-Forest Applied Research Center for Pearl River Delta Environment and Department of Biology, Hong Kong Baptist University, Hong Kong,
Hong Kong Special Administrative Region

a r t i c l e i n f o a b s t r a c t

Article history: The aim of present study was to determine an appropriate ISR (inoculum to substrate ratio) to enhance
Received 8 October 2011 the hydrolysis rate and reduce the solid retention time of food waste in hydrolytic-acidogenesis leach bed
Received in revised form 11 December 2011 reactor (LBR). LBR 1–4 were inoculated with 0%, 5%, 20% and 80% (w/w basis) of anaerobically digested
Accepted 12 December 2011
sludge, respectively, using artificial food waste as substrate. Experiments were conducted in batch mode
Available online 17 December 2011
at mesophilic condition (35 °C) for 17 days. Higher ISR resulted in 4.3-fold increase in protein hydrolysis;
whereas, only a modest increase in the decomposition of carbohydrate. Two kinetic models for carbohy-
Keywords:
drate and protein degradation were proposed and evaluated. The differences among four ISRs in volatile
Acidogenesis
Anaerobic digestion
solids removal efficiencies were marginal, i.e. 52.4%, 62.8%, 63.2% and 71.7% for LBR 1–4, respectively;
Inoculum to substrate ratio indicating that higher ISR was insignificant in enhancing the overall hydrolysis rate in LBR. Therefore,
Kinetics study a lower ISR of 20% was recommended in the hydrolytic-acidogenic process.
Proteolytic bacteria Ó 2011 Elsevier Ltd. All rights reserved.

1. Introduction
The demands for reduction and effective utilization of munici-
pal solid waste (MSW) have increased immensely for Hong Kong
in recent years due to the limited landfill space. Anaerobic diges-
tion of sorted organic fraction of MSW, especially food wastes, is
the utmost attractive alternative and one of the most cost-effective
technologies. Anaerobic digestion (AD) of organic matter is gener-
ally considered to be a two-step process in which the acidogenesis
and methanogenesis are in dynamic equilibrium (Vavilin et al.,
2001). Therefore, it is logical to develop a two-stage anaerobic
digestion system, consisted of leach bed reactor (LBR) and upflow
anaerobic sludge blanket (UASB) that can separate acidogens and
methanogens spatially for the regulation of the two groups of
microbial communities.
In general, the growth rates of acidogens ranged from 0.05 to
1.79 h1, which is about ten times higher than that of acetogens
and methanogens ranging from 0.008 to 0.173 h1 (Kalyuzhnyi,
1997; Vavilin et al., 2001). The imbalance between acid production
by the acidogens and the acid consumption by the methanogens
resulting from the different growth rate is the principle incentive
and momentum of the stage separation. In the start-up of an anaer-
obic digester, an appropriate ratio of inoculum to substrate (ISR)
should be added to enhance the hydrolysis of particular organic
matter, accelerate biogas production rate, and consequently reduce
the operation time (Raposo et al., 2009). About 80% of total biogas
yield was achieved after the first 8 days of digestion for an ISR of
0.35 (vs/vs on the basis of volatile solid) with a single-stage batch
AD of food and green wastes, where it was only 47% for ISR of
0.25 (vs/vs) (Liu et al., 2009).
However, due to the higher growth rate of acidogens than
methanogens (Qi et al., 2003), it is reasonable to expect a lower
ISR in the separated acidogenic reactor than methanogenic reactor.
Nevertheless, previous studies on ISR were mostly investigated in
the single-stage methanogenic reactor (Neves et al., 2004; Liu
et al., 2009), and rarely on the startup of an acidogenic reactor. Fur-
thermore, a wide range of ISR values were employed in existing
studies and it is hard to make a comparison among the different
substrates used. For example, an ISR of 1.0 (vs/vs) was suggested
for the acidogenic fermentation of grass (Jagadabhi et al., 2010),
whereas 0.02 (vs/vs) for manure (Demirer and Chen, 2008) and
0.13 (vs/vs) for food waste (Stabnikova et al., 2008) were reported.
Therefore, the objective of the present study was to determine
the hydrolysis and acidogenesis rate of food waste under different
ISR values, i.e. 0%, 5%, 20% and 80% (w/w) within lab scale LBR at
35 °C. The kinetics of hydrolysis rates of the particulate matters were
then evaluated by the first-order equation and Contois equation.
2. Methods
2.1. Food waste and inoculum

⇑ Corresponding author. Tel.: +852 34117056; fax: +852 34112095. Selected properties of the food waste and inoculum used in the
E-mail address: jwcwong@hkbu.edu.hk (J.W.C. Wong). study are described in Table 1. Synthetic food waste with a total

0960-8524/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2011.12.059
426 S.Y. Xu et al. / Bioresource Technology 126 (2012) 425–430
Table 1
Characteristics of food waste and inoculum.
Bread Boiled rice Cabbage
Proportion in food waste (%) 35 25 25
TS (%) 71.1 29.2 4.6
VS/TS (%) 97.3 99.3 88.1
Total organic carbon (%) 56.2 52.9 41.7
Total nitrogen (%) 2.49 1.16 3.99
solid (TS) content of 38.5% with volatile solid (VS) content of 97.1%
(VS/TS) was used as the substrate. Freshly collected anaerobically
digested sludge (ADS) from the digestion tank of Shek Wu Hui sew-
age treatment plant in Hong Kong was used as the inoculum, with
TS of 3.8% and VS/TS of 85.2%.
2.2. Experimental set-up
Four LBRs with working volume of 4.6 L were used in this study.
Each LBR was provided with a percolating plate at the bottom of
reactor for leachate collection. Four volumes of inoculum, i.e. 0,
50, 200 and 800 g, were tested and each was mixed with 75 g wood
chips (as bulking agent) and 1.0 kg food waste and loaded into each
LBR. In this way, the initial ISR achieved were 0%, 5%, 20% and 80%
(w/w basis), representing LBR1, LBR2, LBR3 and LBR4, respectively,
excluding the weight of wood chips (Table 2). On vs/vs basis, these
ISR ratios correspond to 0, 0.004, 0.017 and 0.069, respectively.
Then 1.0 L of tap water was added into each LBR on day 0. Leaching
occurred naturally and the leachate was collected at the bottom
chamber of the LBR daily. Exactly 50% of the collected leachate
was adjusted to pH 6.0 using sodium carbonate and recycled back
to LBR as the pH adjustment enhanced the chemical oxygen de-
mand (COD) yield in our previous study (Xu et al., 2011). Remain-
ing 50% of the leachates were taken out from LBRs and used for
analysis. The pH, COD, volatile fatty acids (VFA), ammoniacal nitro-
gen (NHþ 4 -N) and total Kjeldahl nitrogen (TKN) of leachate were
determined to evaluate the performance of acidogenic LBRs.
2.3. Analyses
The physicochemical properties including pH, COD, VFA, NHþ4 -N
and TKN of aqueous and solid samples collected were determined
following the Standard Methods of APHA (2005). Due to the filter-
ing effect of sand bed placed at the bottom of LBR, COD of soluble
organic matter represented more than 90% of the total COD in the
collected leachate. Thus, the hydrolyzed product was calculated
based on the total COD of leachate. Experiments were run for
17 days under controlled mesophilic temperature (35 °C) with
two replicates for each ISR. Therefore the reported results are
mean ± standard deviation of two independent analyses. Sigma-
Plot software (version 11.0) was used to create graphs and to per-
form the statistical analysis.
2.4. Kinetics study
First-order kinetics and Contois equation were used in the pres-
ent study to evaluate the hydrolysis rate of food waste in LBR with
varied ISRs. The effects of different processes on the hydrolysis
have traditionally been simplified to the first-order kinetics for
the substrate biodegradation (Vavilin et al., 2008). In the First-or-
der kinetics,
dS
¼ kh S ð1Þ
dt
S is the concentration of particulate substrate; kh represents the
hydrolysis constant (d1 basis). However, a relatively high hydroly-
Pork Food waste mixture Anaerobically digested sludge
15 – –
37.8 38.5 3.8
97.6 97.1 85.2
50.5 56.4 32.2
8.71 4.5 6.6
sis rate was obtained in the anaerobic biodegradability tests with a
high ISR (Fernández et al., 2001), showing some degree of depen-
dence of hydrolysis on biomass concentration or activity. Conse-
quently, the first-order kinetics may not be applicable in all
circumstances.
In the Contois kinetics, hydrolysis is considered to be a surface
reaction occurring when the particulate substrate is in close con-
tact with microorganisms that provide the hydrolytic enzymes
(Tomei et al., 2008). According to the Contois equation, the sub-
strate degradation rate is assumed to be regulated by the relation-
ship of substrate and inoculum, which will be appropriate to
describe the kinetics of hydrolysis of food waste in the LBR with
different ISRs in the present study. Thus, the hydrolysis rates are
evaluated from the mass balance Eq. (2) for substrate and biomass,
respectively,
dS kh S kh  S=X
rh ¼  ¼ X¼X ð2Þ
dt ðK X X þ SÞ ðK X þ S=XÞ
where rh is hydrolysis rate (g CODS d1); kh is hydrolysis constant
(g CODS g CODB1 d1); KX is Contois constant (g CODS g CODB1 d1);
X (g L1) is the concentration of acidogens (responsible for hydroly-
sis of particulate substrate) and S is the particulate substrate; X/S is
the ratio between acidogens concentration (X) and particular sub-
strate (S); CODB and CODS represent the COD content of acidogens
(B) and substrate (S), respectively.
3. Results and discussion
3.1. pH and volume of leachate
pH is an important parameter that can affect the bacterial activ-
ity; and metabolic pathways with a pH of 4.0–6.0 is more favorable
for the hydrolysis and acidogenesis (Li and Fang, 2007). Addition of
inoculum at >20% increased the initial pH of the feedstock to 4.5;
whereas, an initial pH of around 3.8 was observed in 65% inocula-
tion. Daily pH adjustment with sodium carbonate increased the pH
of LBRs gradually from 3.8–4.5 to pH higher than 5.0 after 8 days
(Fig. 1a) as also observed in our previous experiment (Selvam
et al., 2010; Xu et al., 2011). Similar pH changes were noted in
LBR 2–4, with LBR1 exhibited a lower pH in the range of 3.8–5.0.
The lower pH of 3.8 found in LBR1 and LBR2 might be attributable
to the production of formic acid and lactic acid due to their lower
pKa values, i.e. 3.77 and 3.86, respectively. The mixture of acetate,
lactate, formate, succinate and ethanol are the common end prod-
ucts of mixed acid fermentation, which could be mediated by the
members of the family Enterobacteriaceae, Clostridia and lactic
acid bacteria (LAB) (Haruta et al., 2005). LAB were found to be
the preferential bacteria in the kitchen waste that was stored for
24 h at 37 °C (Wang et al., 2001). Meanwhile, the continuous pH
adjustment during the treatment led to the selective reduction of
lactic acid bacteria and increased the coliform bacteria and
Clostridia (Sakai et al., 2000).
The quantities of leachates collected daily from the LBRs were
similar, ranging from 900 to 1100 mL. However, a decrease in
leachate production was observed in the LBR4 after 13 days of
digestion, which would be unfavorable for the soluble organic
 S.Y. Xu et al. / Bioresource Technology 126 (2012) 425–430 427

Table 2
Decomposition of food waste in the LBR with different initial ISRs.

ISR ISR COD leaching rate VFA leaching rate TKN leaching rate VS removal Specific COD yield Specific VFA yield
(w/w, %) (vs/vs) (g kg VSadded1 d1) (g COD kg VSadded1 d1) (g kg VSadded1 d1) efficiency (%) (g g VSadded1) (g COD g VSadded1)
LBR1 0 0 28.1 ± 0.67 3.39 ± 0.13 0.27 ± 0.04 52.4 ± 2.2 0.48 ± 0.01 0.05 ± 0.00
LBR2 5 0.004 30.8 ± 0.58 5.62 ± 0.35 0.48 ± 0.02 62.8 ± 1.9 0.52 ± 0.01 0.09 ± 0.01
LBR3 20 0.017 33.8 ± 1.1 5.99 ± 0.21 0.80 ± 0.11 63.2 ± 0.5 0.56 ± 0.01 0.10 ± 0.01
LBR4 80 0.069 38.8 ± 2.3 10.37 ± 0.68 1.08 ± 0.09 71.7 ± 2.8 0.64 ± 0.07 0.18 ± 0.03

matters leaching out of LBR. The anaerobically digested sludge has
a higher viscosity (0.05 to 0.07 g cm1 s1) than water (Myint et al.,
2010), which might contributed to the increased leaching resis-
tance in LBR. Furthermore, fine particles from the decomposition
of substrate were easily cumulated on the top of filter and hin-
dered the leaching of high-viscosity liquid and consequently led
to the clogging of LBR.
3.2. COD leaching
As seen in Fig. 2a, a biphasic pattern of COD leaching can be ob-
served. COD concentrations of the collected leachate on day 1 were
high ranging from 34.8 g L1 to 47.3 g L1, after that they decreased
gradually. However, the difference of the apparent COD leaching
rates among the four reactors was insignificant during the initial
6 days, i.e. 40.1–42.6 g kg VSadded1 d1 in LBR 2–4, and 36.2 g
kg VSadded1 d1 in LBR1. In the period of day 6–17, the apparent
COD leaching rates were 24.1, 28.1 and 35.1 g kg VSadded1 d1
for LBR 2–4, respectively, which positively correlated with the
amount of inoculum addition. The maximum COD concentration
of >23 g L1 was observed in LBR4; whereas it was less than
15 g L1 in LBR1.
7
Hydrolysis of organic solid waste is frequently evaluated using
first-order kinetics or surface-related Contois model, in which the
hydrolysis rate is proportional to the initial solid waste concentra-
tion, biomass concentration, and the mass-transfer related effi-
ciency (Vavilin et al., 2004). However, a difference in hydrolysis
rate was not observed during the initial stage, which might be
due to a large fraction of water-soluble organic matter existed in
the synthetic food waste. Thus, the high concentration of COD ob-
served during the initial stage might not be originated from the
hydrolysis of particular organic matter. When most of the water-
soluble fraction was washed out of LBR, the effect of inoculum ratio
on the food waste decomposition was observed.
3.3. VFA leaching and acidification degree
Compared with COD, the concentrations of VFA closely reflect
the role of inoculum on the acidogenesis of food waste. During
the first 3 days, VFA concentrations were quite low in the level of
1.2–2.7 g COD L1, which only accounted for 3.0–7.7% of COD as
shown in Fig. 2b. VFA concentrations increased gradually along
with digestion, indicating the growth of acidogenic bacteria in
60
(a)
COD and VFA (gCOD L )
-1

45
(a)
COD
6
30

5
pH

15 VFA
4
0

3
0 3 6 9 12 15 18 60 (b)

1200 (b) LBR1 LBR2

45 LBR3 LBR4
VFA/COD(%)

1000
30
Volume (mL)

800

15
600 LBR1 LBR2
LBR3 LBR4
0
400 0 3 6 9 12 15 18
0 3 6 9 12 15 18 Time (d)
Time (d)
Fig. 2. Leaching of COD and VFA from the LBR with different ISRs: (a) COD and VFA
Fig. 1. (a) pH and (b) volume of leachates from the LBRs with different inoculum to concentrations, (b) ratio of VFA to COD (ISR = 0% (LBR1), 5% (LBR2), 20% (LBR3) and
substrate ratio (ISR) values (ISR = 0% (LBR1), 5% (LBR2), 20% (LBR3) and 80% (LBR4)). 80% (LBR4)).
428 S.Y. Xu et al. / Bioresource Technology 126 (2012) 425–430

LBR with different levels of VFA being observed among the four
LBRs which reflected different sources of inoculum or ISR. The
highest concentration of VFA observed in LBR4 was 11.8 g COD L1,
whereas it was only 6.8 g COD L1 in LBR1. Correspondingly, the
overall VFA leaching rates decreased from 10.37 g kg VSadded1 d1
of LBR4 to 3.39 of g kg VSadded1 d1 of LBR1.
Acidogenesis was reported to be determined primarily by the
hydrolysis rate rather than by the bacterial growth kinetics, due
to the rate limiting step of hydrolysis (Eastman and Ferguson,
1981; Vavilin et al., 2001). Nevertheless, the soluble COD in present
study was quite high when compared to VFA. Meanwhile, metha-
nogens, the consumer of VFA, should be successfully suppressed
in the reactor due to the low pH values (Dogan et al., 2009). Thus,
under this situation, the leaching rate of VFA, the major end prod-
ucts of acidogenesis, was more correlated to the acidogens popula-
tion than the hydrolysis rate. As shown in Table 2, a higher
acidogenesis rate was observed in the LBR with a higher ISR. The
total VFA yield from LBR4 was 0.18 g kg VSadded1, which was
3.6-fold higher than LBR1 (0.05 g kg VSadded1) and two-fold higher
than LBR2 (0.09 g kg VSadded1).
The degree of acidification (%) in the LBR was calculated from
the VFA yield and COD input. A partial acidification (about 20–
40%) is reported to stimulate significantly the activity of a metha-
nogenic reactor placed in series with the hydrolysis-acidogenic
reactor (Lettinga and Hulshoff Pol, 1991; Guerrero et al., 1999);
suggesting that the optimum conditions in the first reactor should
aim for the maximum acid production rate. Overall tVFA produced
at the end of digestion from LBR 1–4 were accounting for 6.2%,
10.4%, 10.7% and 20.7% of the total COD input, respectively. It
was comparable to the values obtained in similar LBR studies
(Dogan et al., 2009; Jagadabhi et al., 2011), but relatively lower
than the 50–80% reported in a continuous stirred tank reactor
(Cavinato et al., 2011).
As described previously, the initial high rate of COD leaching
was partially attributed to the water-soluble fraction of food waste
itself, which resulted in a lower degree of acidification initially, i.e.
2–8%. As the readily-soluble COD in LBR decreased, the ratio of
VFA/COD concomitantly increase to 24–46% toward the end of
digestion (Fig. 2b). Furthermore, acidification degree in present
study did not cover the metabolites like formic acid, lactic acid,
alcohols (ethanol, methanol, buthanol, etc.), ketones (glycerol, ace-
tone, etc.), CO2 and H2 generated from different metabolic path-
ways that may explain the low degree of acidification. As
discussed in Section 3.1, the low pH values (3.8) indicated the
production of lactic acid and formic acid in LBR1 and LBR2. How-
ever, those two compounds were not measured and not used for
the calculation of acidification degree.
3.4. VFA speciation
The variations of VFA speciation in LBRs with different ISRs are
presented in Fig. 3. Among the 4 LBRs with different ISRs, acetate
and butyrate were the pre-dominant VFA species throughout the
digestion. High concentration of acetate was produced from the
first day of digestion, while butyrate increased gradually and
peaked at the middle or later period of the experiment. The highest
concentrations of butyric acid were 4.0–5.4 g COD L1, which was
in a similar level as reported by others (Komemoto et al., 2009;

15 15
(a) LBR1 (b) LBR2

12 12
VFA(gCOD L-1)

9 9

6 6

3 3

0 0
1 2 4 6 8 10 13 16 1 2 4 6 8 10 13 16
15 15
(c) LBR3 (d) LBR4

12 12
VFA(gCOD L-1)

9 9

6 6

3 3

0 0
1 2 4 6 8 10 13 16 1 2 4 6 8 10 13 16
Time (d) Time (d)

Acetic acid Propionic acid Iso-butyric aicd Butyric acid

Iso-valeric acid Valeric acid
Fig. 3. Variations of VFA composition in leachates from the LBRs with different ISRs (ISR = 0% (LBR1), 5% (LBR2), 20% (LBR3) and 80% (LBR4)).
 S.Y. Xu et al. / Bioresource Technology 126 (2012) 425–430 429

Jagadabhi et al., 2010). Acid-forming pathway with acetate and

butyrate as the major metabolites could be favorable for the suc- 1200
(a)
ceeding methanogenesis, setting biogas as the end product in
two-phase anaerobic digestion (Horiuchi et al., 2002).

TKN (mg L )
900

-1
A relatively high concentration of propionate was only occurred
in LBR4, ranging from 0.40 to 2.92 g COD L1, while it was less than
600
1.0 g L1 for the other three LBRs similar to that observed by
Komemoto et al. (2009). Hawkes et al. (2002) also found the conver-
sion of acetate and butyrate to propionate production as the pH 300
increased, which might be attributed to a change in the dominant
microbial population, or change in the metabolic pathway within 0
the same bacterial population. Propionate higher than 35 mM might
inhibit the acetoclastic methanogenesis (Van Lier et al., 1993), 600
which needs to be avoided in the methanogenic reactor. LBR1 LBR2 (b)
500 LBR3 LBR4
3.5. Leaching of TKN and NHþ

NH4 -N (mg L )
4 -N

-1
400

The breakdown of protein-rich organic matter results in the re- 300

lease of relatively large amounts of NHþ 4 -N by hydrolysis of protein

+
and ammonification of amino acids (Venkata Mohan et al., 2008). 200
The leaching pattern of TKN and NHþ 4 -N are presented in Fig. 4.
100
On the 1st day, the TKN concentrations in the collected leachate
were almost similar among four LBRs. This correlated well with 0
the trend of COD leaching indicating that the initial leaching rate 0 3 6 9 12 15 18
of organic matter from the LBR was mainly related to the water- Time (d)
soluble fraction of food waste rather than that of ISR. Similarly,
the soluble NHþ 4 -N in the inoculum contributed to the initial high
Fig. 4. (a) TKN and (b) NHþ 4 -N concentrations in leachates from the LBRs with

concentration of ammonia in the leachate. Meanwhile, although different ISRs (ISR = 0% (LBR1), 5% (LBR2), 20% (LBR3) and 80% (LBR4)).

the ratio of inoculum addition in LBR4 was high (80% on weight ba-
sis), the N content from ADS was less than 10% of that from food
waste due to the low TS content of ADS (3.8%) compared to food
waste (38.5%).
As shown in Fig. 4, LBR4 with the highest ISR resulted in the
highest levels of TKN and NHþ 4 -N, while LBR1 produced the lowest
concentrations demonstrating the decomposition of protein clo-
sely related to ISR in the later stage. The apparent TKN leaching
rates were 0.27, 0.48, 0.80 and 1.08 g kg VSadded1 d1 for LBR 1–
4, respectively (Table 2), indicating that a higher ISR leads to a
higher hydrolysis rate of protein due to proteolytic bacteria (Sie-
bert and Toerien, 1969), which was observed in a high population
in anaerobically digested sludge. However, the proteolytic organ-
isms were apparently washed progressively at pH below 6 (Erfle
et al., 1982), which led to the concomitant decrease of TKN and
NHþ 4 -N in leachate. Especially for LBR1, the concentrations of TKN
and NHþ 4 -N decreased to less than 200 mg L
1
and 10 mg L1 after
5 days of digestion, indicating the inactive state of proteolytic en-
zymes and ammonification. The low concentration of NHþ 4 -N also
correlates well with the low buffering capacity and low pH values
in LBR1, as seen in Fig. 1a. The results support the addition of active
inocula into the hydrolytic reactor to enhance protein degradation.
3.6. Kinetics evaluation of the hydrolysis and acidogenesis rate of food
waste
COD and TKN degradation data were analyzed using first-order
kinetics equation and Contois equation. The fitting of correlation
was evaluated with the correlation coefficient R2, and the reliabil-
ity of the evaluated parameters was quantitatively estimated by
standard error (SE).
As presented in Table 3, a modest increase of overall hydrolysis
rate as indicated by the decrease in COD was observed from 0.042
to 0.069 d1, when increasing ISR from 0 to 0.069 (vs/vs). The cor-
relation of COD hydrolysis rate and ISR was insignificant (p > 0.05).
Nevertheless, a similar hydrolysis rate (Kh) of 0.86–1.11 g CODS g
CODB1 d1 was observed when including the effect of X/S in the

Table 3
Kinetic parameters for the anaerobic digestion of food waste at different ISRs.

First-order equation ISR kh (d1) SE R2

COD 0 0.042 9.19E04 0.99
0.004 0.056 9.56E04 0.99
0.017 0.058 1.44E03 0.99
0.069 0.069 2.66E03 0.98
First-order equation ISR kh (d1) SE R2

TKN 0 0.006 2.65E04 0.97

0.004 0.011 7.93E04 0.93
0.017 0.023 4.33E04 0.99
0.069 0.032 7.14E04 0.99
Contois equation ISR Kh (g CODS g CODB1 d1) KX (g CODS g CODB1 d1) R2
COD 0 1.11 18.05 0.99
0.004 1.08 25.35 0.99
0.017 0.95 31.15 0.98
0.069 0.86 62.31 0.99
430 S.Y. Xu et al. / Bioresource Technology 126 (2012) 425–430
Contois equation, which indicates that the bioavailability of COD of
food waste is quite high and the activity of newly produced bacte-
ria were comparable to the bacteria from the initially added inoc-
ulum. However, it is worth to note that a minor decrease of Kh
appeared in LBR4, which might be due to the effect of the higher
amount of the inert fraction in the inoculum, the degradation of
which also contribute to the degradation rate of substrate. Similar
result was also reported by Tomei et al. (2008).
The hydrolysis rate of protein increased from 0.006 to 0.032 d1
with an increase in ISR, which is comparatively lower than the
overall hydrolysis of COD. The experimental results of Henze and
Mladenovski (1991) indicated that the hydrolysis of nitrogenous
compound was 0.06 and 0.03 d1 under anaerobic and anoxic con-
ditions. Nevertheless, the protein hydrolysis was rate limiting un-
der acidic conditions (Xu et al., 2011), so that the observed kinetics
is strongly limited by the substrate availability and the activity of
inoculum.
As shown in Table 2, VS removal efficiencies (%) of LBR 1–4 were
52.4, 62.8, 63.2 and 71.7, respectively. The VS removal efficiencies
were insignificantly positively correlated with ISR (r = 0.873,
p = 0.064). The modest increase of VS removal efficiencies in the
LBR with higher ISR was an evidence of the insignificant effect of
ISR on the initial liquefaction rate and biodegradability of food
waste in LBR. Meanwhile, TKN leaching rates showed a strong sig-
nificant positive correlation with ISR (r = 0.918, p < 0.05). Never-
theless, the relatively low amount of protein in food waste (11%
of TS) made protein hydrolysis contribute little to the overall
decomposition rate. On the other hand, if the ISR was too high, it
would reduce the capacity of the reactor due to the large volume
of inoculum should be added. In view of that, a low ISR ratio of
<20% would be enough to obtain a fast acid production in the aci-
dogenic reactor during the start-up of a two-phase anaerobic
system.
4. Conclusions
With an increase in ISR, the kinetics of liquefaction and acidifi-
cation of food waste decomposition in the LBR were increased with
evidence from the analysis using the first order kinetics and Con-
tois equation. Despite the higher COD and VFA production with
>20% ISR, the increase in food waste decomposition was not signif-
icant. Instead, it reduced the process efficiency of food waste treat-
ment and increased the demand for high inoculum size. Thus a
lower ISR of 20% (w/w) is recommended for the hydrolytic-
acidogenic process of food waste in view of the cost effectiveness
and decomposition efficiency.
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