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Water Research 36 (2002) 43694385

Comparative process stability and efciency of anaerobic


digestion; mesophilic vs. thermophilic
Moonil Kima,*, Young-Ho Ahnb, R.E. Speecea
a
Department of Civil & Environmental Engineering, Vanderbilt University, PO Box 1831, Station B, Nashville, TN 37235, USA
b
Department of Civil Engineering, Yeungnam University, Kyungsan 712-945, South Korea
Received 1 August 2001; received in revised form 1 March 2002; accepted 19 March 2002

Abstract

The comparative process stability and efciency of mesophilic (351C) and thermophilic anaerobic digestion (551C)
has been evaluated for four different reactor congurations, which are: daily batch-fed single-stage continuously stirred
tank reactor (CSTR), continuously fed single-stage CSTR, daily batch-fed two-phase CSTR, and daily batch-fed non-
mixed single-stage reactor. The results are discussed for three periods: (1) start-up, (2) steady state, and (3) organic
loading rate (OLR) increase until reactor failure (pH below 5.5). During the start-up, the single-stage CSTRs at both
temperatures showed the least stability, while the non-mixed single-stage reactors reached steady state in the shortest
time with relatively stable pH and low volatile fatty acid (VFA). In the case of the thermophilic non-mixed reactor,
efcient removal of propionate occurred but supplementation of nutrients (Ca, Fe, Ni, and Co) was required when
VFA increased. The results imply the importance of inorganic nutrients bioavailability. The comparative results of the
reactor performance at steady state clearly showed the superior performance of the thermophilic non-mixed reactor
with respect to lower VFA, higher gas production and volatile solids removal implying that microbial consortia
proximity can alleviate the problem of poor efuent quality in thermophilic system. During the OLR increase until
reactor failure, all thermophilic reactors except the thermophilic non-mixed reactor showed increases in propionate
concentrations as the OLR increased, while all mesophilic reactors except the mesophilic two-phase system showed little
increase in VFA concentrations. When all reactors had the same conditions with OLR increase, the continuously fed
reactors showed the lowest gas production, while the non-mixed reactors showed the highest gas production at both
temperatures. It is hypothesized that the non-mixing reactor conguration has closer microbial consortia proximity
than others. Therefore, the results in this study indicated the importance of microbial consortia proximity. A proposed
model for the effect of the distance between two syntrophic bacteria reasonably matched the data in this study. r 2002
Elsevier Science Ltd. All rights reserved.

Keywords: Mesophilic; Microbial consortia proximity; Nutrients; Reactor conguration; Thermophilic; Volatile fatty acid

1. Introduction destruction rate of organic solids, improved solids


liquid separation, and increased destruction of patho-
Mesophilic temperature regimes (30401C) have long genic organisms [13]. However, the use of thermophilic
been adopted for anaerobic digestion, showing good anaerobic digestion has been limited, because of some
operational performance. Thermophilic regimes (50 disadvantages like poor supernatant quality and poor
601C) have also been adopted for anaerobic digestion process stability related to chronically high propionate
showing several advantages, such as an increased concentrations [4].
It has been shown by previous studies that thermo-
*Corresponding author. Tel.: +1-615-322-4348. philic microorganisms are characterized by their higher
E-mail address: moonil.kim@vanderbilt.edu (M. Kim). substrate utilization and growth rates as well as higher

0043-1354/02/$ - see front matter r 2002 Elsevier Science Ltd. All rights reserved.
PII: S 0 0 4 3 - 1 3 5 4 ( 0 2 ) 0 0 1 4 7 - 1
4370 M. Kim et al. / Water Research 36 (2002) 43694385

decay rate compared to mesophilic bacteria. Studies at H2 concentrations between 104 to 106 atm. This
undertaken by several researchers [57] showed that is a severe limitation and an example of exquisite
thermophilic systems were capable of treating higher microbial ecological partnership. Diffusion of dis-
organic loadings and had higher specic growth rate as solved H2 from the microorganisms that produce it
compared to their mesophilic counterparts. The yield of to the microorganisms which consume it is the main
these microorganisms per unit amount of substrate is mechanism for processes on this microscale and
lower. The lower growth yield of thermophilic bacteria therefore close microbial proximity favors low H2
could be due to their increased decay rate, which is concentrations [1012].
double that of mesophilic cultures because the cells have * Limit of nutrient bioavailability: Essential in micro-
a tendency to lyse quickly under thermophilic conditions and macroinorganic nutrients even though at present
[8], and may be due to their higher energy requirement it may not be sufciently bioavailable to maintain
for maintenance or the specic molecular properties of efcient enzymatic processing.
enzyme reactions at thermophilic temperatures [9].
The purpose of this research is to evaluate the
Class A biosolids termed by EPA are essentially
comparative process stability and process efciency of
pathogen free and have no restrictions on crop choice or
thermophilic vs. mesophilic anaerobic digestion by
human access to land application sites used for disposal.
evaluating the key parameters of reactor conguration,
Mesophilic anaerobic digestion cannot meet the criteria
microbial consortia proximity, and nutrient supplemen-
of Class A biosolids because of insufciently high
tation. For the sake of comparison, all experiments in
temperatures. Thermophilic anaerobic digestion is one
this research were conducted at 551C or 351C so that
of the proven methods for producing Class A biosolids.
thermophilic operation can be directly compared with
Even though thermophilic anaerobic digestion has
the more commonly used mesophilic temperature.
proven pathogen reduction potential, US EPA (technol-
ogy-based, 40 CFR 257, 1989) excluded thermophilic
digestion from processes to further reduce pathogens
(PFRP) and US EPA (performance-based, 40 CFR 503, 2. Materials and methods
1992) listed aerobic thermophilic digestion as just one of
the PFRP alternatives, still not including single-stage 2.1. Seed biomass, substrate, and temperature
anaerobic thermophilic digestion because of instabil-
ity in operation and possibility of short circuiting The thermophilic seed source was taken from a
characteristics of single-stage reactor [14,15]. thermophilic anaerobic sludge culture, which was
Thermophilic anaerobic digestion often manifests enriched on calcium acetate (10,000 mg/L) and calcium
chronically higher volatile fatty acid (VFA) concentra- propionate (4000 mg/L) for 3 months in a 17 L batch-fed
tion (especially propionate) than that for mesophilic. reactor at a temperature of 551C with an HRT of 30
Three hypotheses are put forth to explain this situation: days. This culture was originally taken from the
Murfreesboro, TN, municipal anaerobic sludge digester
* Sinlge-stage continuously stirred tank reactor which had been operated at mesophilic temperature. The
(CSTR) configuration: In conventional, single-stage mesophilic seed source was also taken from the same
CSTR, anaerobic digestor, all phases (i.e. hydrolysis, digester. The substrate was a simulated primary sludge.
acidogenesis, and methanogenesis) take place in a Dog food was selected for the substrate due to its
dispersed common environment which is not ideal for similarity to municipal primary sludge. This material
all members of consortia. had protein, fat, and ber contents of 21%, 8%, and
* Syntrophic reaction: A consortia of up to three 5%, respectively, and a volatile solid (VS) content of
groups of bacteria are involved as shown in Fig. 1. 90% of total solids (TS). It also contained all the micro-
Propionate conversion to acetate/H2 and H2 conver- and macronutrients necessary for animal growth. The
sion to methane is thermodynamically possible only feed slurry was prepared by blending after mixing with

Fig. 1. Three groups of bacteria involved in propionate degradation.


M. Kim et al. / Water Research 36 (2002) 43694385 4371

tap water containing 6 g/L of NaHCO3. Temperatures (Beckman Instruments, Model GP) at 4000 rpm for
of 551C and 351C were selected as the thermophilic and 30 min and ltering approximately 3 mL of the super-
mesophilic temperatures, respectively. natant through 0.45 mm lter paper (Micron Separations
Inc.) The ltered samples were then acidied with 10%
2.2. Apparatus H3PO4 (volumetric basis) to lower the pH below 3 and
ensure that acids were unionized and able to volatilize.
Four different reactor congurations, which are a VFAs were measured twice per week. TS were measured
daily batch-fed single-stage CSTR, a continuously fed according to section 2540 D and VS were measured
single-stage CSTR, a daily batch-fed two-phase CSTR, according to section 2540 E in Standard Methods,
and a daily batch-fed non-mixed reactor, were studied at respectively [13]. TS and VS were measured twice per
551C and at 351C, respectively. Table 1 tabulates a week.
summary of operating conditions evaluated and Fig. 2
shows the schematic of these four different reactor
congurations. MB, MC, MTP2, MNMR, TB, TC, 3. Results and discussion
TTP2, and TNMR utilized 1.1 L glass bottles (Wheaton
Scientic Co.) and the operating volume was 800 mL. The results are discussed for three periods: (1) start-up
MB and TB were fed once per day and MC and TC were until steady state (Days 0200); (2) steady state with feed
fed continuously. MB, MC, MTP2, TB, TC, and TTP2 solids concentration of 4% and HRT of 20 days (Days
were continuously mixed by a magnetic stirrer. MTP1 201248); and (3) organic loading rate (OLR) increase
and TTP1 utilized 500 mL glass bottles and the from 4% solids at steady state to reactor failure by
operating volume was 120 mL. MTP1, MNMR, TTP1 increasing the solids concentration in the feed and
and TNMR were mixed manually just prior to wastage decreasing the HRT (after Day 201).
and immediately after feeding and then allowed to settle
and remain unmixed until the next days feeding. 3.1. Start-up (Days 0200)

2.3. Analytical methods Before starting operation of all reactors except the
TNMR, it was intended to compare reactor perfor-
Daily gas production in all reactors was measured mance at the feed solids content of 4% and HRT of 20
using wet-tip gas meters. pH in all reactors was days. However, during the rst 4% feeding period all
measured daily using a pH meter (Fisher Co. digital reactors showed a pH drop as well as an increase of
pH meter 107). VFAs in all waste samples were VFA concentration. Therefore, the solids percent in the
measured using a gas chromatograph (Shimadzu Model feed was reduced to 0% or 2% and then increased
GC-14A) equipped with a ame ionization detector and gradually up to 4% depending on the pH recovery of
a 1.7 m glass column packed with 0.3% GP Carbopack each reactor. Stable reactor performance of all reactors
C/0.3% Carbowax 20 M/0.1% H3PO4 (Supelco, Inc.). reached 4% of feed solids percent at different intervals
The column temperature was 1401C and the injector/ (Fig. 3), which were:
detector temperature was 2001C. Nitrogen was the
carrier gas with a pressure of 1.75 kg/cm2. Data * MB at Day 178;
integration was accomplished using a Varian 4270 * TB at Day 178;
Integrator. Samples were prepared by centrifuging * MC at Day 63;

Table 1
Summary of reactor operating conditions

Conguration Temperature (1C) Starting HRT (days) Feed

TB: thermophilic batch-fed CSTR 55 20 Dog food


TC: thermophilic continuously fed CSTR 55 20 Dog food
TTP1: phase 1 reactor in thermophilic two-phase system 55 2 Dog food
TTP2: phase 2 CSTR in thermophilic two-phase system 55 18 Efuent TTP1
TNMR: thermophilic batch-fed non-mixed reactor 55 30 Dog food
MB: mesophilic batch-fed CSTR 35 20 Dog food
MC: mesophilic continuously fed CSTR 35 20 Dog food
MTP1: phase 1 reactor in mesophilic two-phase system 35 2 Dog food
MTP2: phase 2 CSTR in mesophilic two-phase system 35 18 Efuent MTP1
MNMR: mesophilic batch-fed non-mixed reactor 35 20 Dog food
4372 M. Kim et al. / Water Research 36 (2002) 43694385

Fig. 2. Schematic of four reactor congurations.

* TC at Day 63; With respect to the comparison of temperature effect, all


* MTP at Day 56; reactor congurations at thermophilic temperature
* MNMR at Day 49; and except the two-phase system (i.e. all single-stage
* TTP at Day 44. reactors) showed higher acetate concentrations (Fig. 3).
Even though individual specic microorganisms (i.e.
TNMR started with an HRT of 30 days at the feed acidogens and methanogens) at thermophilic tempera-
solids content of 4% and the HRT was changed to 20 ture have a higher substrate utilization rate, these results
days at Day 120. During the period before adding implied that slower growing methanogens in a single-
nutrients, both MB and TB were not only less stable stage were more affected by the higher temperature,
than other reactors but also reached 4% feed solids the probably due to an unbalanced reaction rate between
latest (Fig. 3). This result appears to indicate that, for acidogens and methanogens. On the other hand, for the
the start-up period: two-phase reactor conguration, no difference between
* two-phase had an advantage over single-stage from the two temperatures was observed until supplementing
the favorable results of MTP2 and TTP2; nutrients, probably due to the separation of the two
* continuous feed had an advantage over batch feed kinds of microorganisms. Decreasing the HRT of
from the favorable results of MC and TC; and TNMR from 30 to 20 days did not affect the reactor
* non-mixing had an advantage over CSTR from the performance (Figs. 3, 4 and 6).
favorable results of MNMR. Fig. 5 shows the results of start-up for both phase 1
reactors in the two-phase system (MTP1 and TTP1) with
Measured VFA compounds and organic acids were respect to VFA and pH vs. time and average results
acetate, propionate, iso-butyrate, n-butyrate, 2-methyl- from Days 64 to 200 (i.e. HRT of 2 days and feed
butyrate, iso-valeric, n-valeric, and lactate. Among them solids of 4%). Both phase 1 reactors had high concen-
the main acids were acetate, propionate, and n-butyrate. trations of:
M. Kim et al. / Water Research 36 (2002) 43694385 4373

Fig. 3. Monitoring start-up of reactors for pH and acetate.


4374 M. Kim et al. / Water Research 36 (2002) 43694385

Fig. 4. Monitoring start-up of reactors for propionate and n-butyrate.


M. Kim et al. / Water Research 36 (2002) 43694385 4375

Fig. 5. Monitoring start-up of MTP1 and TTP1 for VFA and pH. (a) VFA and pH vs. time. (b) Average from Days 64 to 200. (Error
bar represents 2 standard deviations.)

* acetate (mesophilic: 20005000 mg/L; thermophilic: was added to all reactors having manifested chronically
10003000 mg/L); elevated VFA concentrations (i.e. all reactors except
* n-butyrate (mesophilic: 100010,000 mg/L; thermo- MNMR) on Day 69.
philic: 10007000 mg/L); and Added inorganic nutrients consisted of CaCl2, FeCl2,
* lactate (mesophilic: 200011,000 mg/L; thermophilic: CoCl2, and NiCl2, at the concentration of 500, 10, 1,
10007000 mg/L). and 1 mg/L respectively. Soon after adding nutrients
daily, the gas production of those reactors increased
But propionate was barely produced (o100 mg/L) at signicantly followed by a slight decrease and, there-
both temperatures. On the other hand, the phase 2 after increased gradually until steady state (Fig. 6).
reactors produced high levels of propionate (mesophilic: Even though the substrate (dog food) had enough
before adding nutrients (02000 mg/L), after adding nutrients for animals, the positive results from nutri-
nutrients (20006000 mg/L); thermophilic: no difference ent supplementation show that these reactors were
between before and after adding nutrients (1000 decient in bioavailability of those nutrients at
2000 mg/L)). The pH of TTP1 was maintained at a both temperatures. The feed contained more than
slightly higher range (between 5 and 5.5) than that of ample nutrients but its bioavailability was denitely
MTP1 (between 4.5 and 5) (Fig. 5). limiting probably due to precipitation reactions
In order to observe the effect of inorganic nutrient in the reactor, in all probability. Therefore, the results
supplementations, a cocktail of Ca, Fe, Co, and Ni, imply the importance of inorganic nutrients bioavail-
which were mostly considered as stimulating nutrients, ability.
4376 M. Kim et al. / Water Research 36 (2002) 43694385

Fig. 6. Monitoring start-up of reactors for gas production and VS destruction (gas production: 4-day moving average).
M. Kim et al. / Water Research 36 (2002) 43694385 4377

Soon after supplementing inorganic nutrients, the thermodynamics of the propionate conversion. Propio-
pH of all reactors at both temperatures increased nate concentrations below 50 mg/L were also the most
for a short time and then decreased below the pH signicant characteristic of MNMR and TNMR. With
before adding nutrients. Normally increased gas produc- respect to the poor efuent quality at the thermophilic
tion accompanies increasing pH because methano- temperature, it turned out that only close microbial
genesis reduces VFA and produces alkalinity. However, consortia proximity could solve this problem during
the results in this experiment show a pH reduction start-up period.
with increased gas production. This result may be
explained by one hypothesis, which is that nutrients 3.2. Steady state with feed solids concentration of 4% and
increased the rate of acidogenesis as well as methano- HRT of 20 days
genesis. The stimulated acidogenesis produced more
VFA and H+ ions, then also stimulated methanogen- For the comparison of the various reactor perfor-
esis, which removed more VFA, but did not produce mances at the steady state with a feed solids concentra-
enough alkalinity to neutralize the H+ ion. These tion of 4% and an HRT of 20 days, all data were
phenomena occurred after the supplementation of calculated for the average value with 1 standard
nutrients and continued until reactor failure. The deviation from Days 200 to 248. During this period,
average pH values are compared later in this the MNMR was not supplemented by inorganic
study. nutrients because unlike the other three reactor cong-
Adding inorganic nutrients signicantly removed urations, MNMR manifested no elevated VFA. The
almost all VFA, except propionate, in all reactors except calculated averages are tabulated in Table 2.
the non-mixed reactor. Residual propionate concentra- The comparisons of average values are shown in
tion was little changed or even increased after supple- Fig. 7. Fig. 7(a) shows that most VFAs consisted of
menting inorganic nutrients. However, in the TNMR, propionate in all reactors. Thermophilic single-stage
propionate was removed to below the detection limit at CSTRs produced much higher VFA concentrations than
the HRT of 30 days and even 20 days (Fig. 4). The their mesophilic counterparts. The two-phase reactor
complete reduction of propionate soon after supple- conguration had much reduced performance over the
menting nutrients can be explained by three parameters, single-stage reactor type at mesophilic temperature while
which are longer HRT, inorganic nutrient supplementa- two-phase had a slight benet over the single-stage
tions, and closer microbial consortia proximity. At the reactor type at thermophilic temperatures. On the other
HRT of 30 days, it is hard to say which parameter or hand, the thermophilic non-mixed reactor showed much
what combination of these three parameters caused this lower VFA concentrations among all thermophilic
result. However, no increase of propionate concentra- reactors as it also did during the start-up period. This
tion after 3 HRT of operation with the HRT of 20 days result implies that the problem of the poor efuent
implies that closer microbial consortia proximity quality at thermophilic temperature can be at least
strongly favored the removal of propionate. In addition, partly solved by non-mixing (i.e. closer microbial
nil propionate concentration in the MNMR also consortia proximity).
explains the same reason. Fig. 7(b) shows the comparisons of VS and VSS
In order to stimulate propionate metabolism, a removal percentage. TB showed higher VSS removal
number of assays were conducted, in which yeast ability than MB while no difference was observed for VS
extract, NH4Cl, (NH4)2HPO4, Na2SO4, MgSO4, KCl, removal between these two reactors. This result implies
ZnCl2, Na2WO4, Na2MoO4, Na2SeO3, MnCl2KI, that more solubilized compounds (i.e. VFAs) at the
NH4VO3, AlCl3, H3BO3, and CuCl2, were assayed in thermophilic temperature were not converted to CH4 as
aliquots taken from reactors showing chronically efciently. Therefore, the results clearly show the higher
elevated propionate concentrations (i.e. MB, MTP2, rate of hydrolysis/acidogenesis and lower rate of
TB and TTP2). However, no stimulation in any of the methanogenesis at the thermophilic single-stage batch-
assays for the degradation of propionate was observed fed CSTRs. On the other hand, TNMR showed higher
at both temperatures (data not shown). VS removal than the thermophilic single-stage CSTRs
Comparing all reactor congurations at both tem- (TB and TC), which implies that the solubilized
peratures, the MNMR and TNMR appeared to show compounds (i.e. VFA) were converted to CH4 ef-
markedly superior performance during the start-up ciently. Therefore, this result implies that in order to
period. This was most probably related to closer raise the rate of methanogenesis at thermophilic
microbial consortia proximity, which minimizes the temperature, closer microbial consortia proximity is
distance between H2 producing and H2 consuming necessary.
microbes, removing the H2 diffusion bottleneck of the Fig. 7(c) shows the comparison with respect to pH.
biotransformation and thus facilitating the scavenging Only MNMR continued to show a higher pH range
of H2 to low concentrations which enhanced the among all reactors. However, even though this higher
4378 M. Kim et al. / Water Research 36 (2002) 43694385

Table 2
Results of reactor performance at steady state operation

Reactor pH Gas production Propionate VFA VS VSS


conguration (mL/g VS (COD mg/L) (COD mg/L) removal removal
added day) (%) (%)

MB Ave. 6.84 494 56 142 68 67


S.D. 0.10 32 25 62 1 1
MC Ave. 6.99 482 46 129 76 76
S.D. 0.04 36 21 58 1 1
MTP Ave. 6.93 531 4495 5105 71 82
S.D. 0.21 57 1184 1316 3 1
MNMR Ave. 7.55 556 56 161 74 74
S.D. 0.08 37 41 90 1 1
TB Ave. 6.76 391 2113 4307 66 76
S.D. 0.06 37 239 835 1 1
TC Ave. 6.89 461 2995 4133 71 80
S.D. 0.06 29 239 546 1 2
TTP Ave. 6.95 607 1859 2444 77 81
S.D. 0.05 37 249 747 2 1
TNMR Ave. 6.94 716 53 162 77 77
S.D. 0.11 42 34 111 1 1

Operating period: from Day 200 to Day 248.


No. of observation: pH (49), Gas production (49), Propionate (7), VFA (7), VS (6), VSS (6).
VS removal %: (VS CODinVS CODout)/VS CODin.
VSS removal %: (VSS CODinVSS CODout)/VSS CODin=(VSS CODin(VS CODoutVFA CODout))/VSS CODin assuming that
most soluble compounds consisted of VFAs.

Fig. 7. Comparison of mesophilic and thermophilic reactors for (a) VFAs, (b) VS and VSS destruction, (c) pH, and (d) gas production.
(Average values during Days 200248, steady state operation with Feed solids %=4 and HRT=20 days, except no nutrients for
MNMR; error bars stand for 2 positive and negative standard deviations.)
M. Kim et al. / Water Research 36 (2002) 43694385 4379

Fig. 8. Long-term monitoring for the failure pattern of all reactors by pH drop.

range of pH may give a good environment for content was observed among all reactors at both
methanogens, it turned out that inorganic nutrient temperatures.
supplementation was perhaps necessary at the higher
OLR because MNMR eventually failed at a lower 3.3. Long-term operation with OLR increase
OLR than three other reactor congurations in spite of (after Day 201)
the advantage of non-mixing (i.e. closer microbial
consortia proximity) (Fig. 8). Fig. 7(d) shows the The operation of all reactors was then continued until
comparison with respect to the average gas production. reactor failure (pHo5.5) in the third phase of this study.
Among all reactors, TNMR produced the highest After a feed solids concentration of 9% was achieved,
unit volume of gas while TB produced the least. Among further increase in OLR was achieved by a reduction of
all mesophilic reactors, no difference in gas production HRT, while maintaining the feed solids at 9%. Table 3
was observed while a severe difference among thermo- shows the OLR at reactor failure and possible positive
philic reactors was observed. These results imply that and negative reasons for the failure.
even though thermophilic bacteria have a higher Fig. 8 shows the failure pattern of all reactors as
activity, the reactor performance strongly depends on manifested by pH drop. As the OLR increased, all
the reactor conguration. All reactors had CH4 compo- reactors eventually failed. TB failed the rst while MC,
sitions of 5560%. No statistical difference in methane TC and TNMR failed the last at the highest OLR. The
4380 M. Kim et al. / Water Research 36 (2002) 43694385

Table 3
The comparison of reactor conguration for failure with OLR increase

Reactor conguration The order of failure OLR (g VS/L day) at failure Possible positive reasons Possible negative reasons

TB 1st 7.4 (4) (1), (2), (3)


MNMR 2nd 7.4 (1), (2) (3), (4)
TTP 3rd 7.4 (1), (4) (2), (3), (5)
MB 4th 10.1 (2), (4) (1), (3)
MTP 4th 10.1 (1), (2), (4) (3), (5)
MC 6th 20.3 (2), (3), (4), (5) (1)
TC 6th 20.3 (3), (4), (5) (1), (2)
TNMR 6th 20.3 (1), (4) (2), (3)

Possible positive reasons: (1) good microbial consortia proximity; (2) faster yield of methanogens due to low temperature; (3) constant
pH due to continuous feeding; (4) nutrients supplementation; (5) waste of non-acidied VS in feed.
Possible negative reasons: (1) poor microbial consortia proximity; (2) slower yield of methanogens due to high temperature;
(3) difculty of pH recovery due to batch feeding; (4) limited bioavailability of nutrients; (5) 2 days shorter HRT/STR.

continuously fed reactors had a relatively constant pH at at 4% feed solids) to Day 345 (1 day before increasing
steady state. However, the pH of daily batch-fed the OLR to 7.4 g VS/L day, when the rst reactor failed),
reactors dropped soon after feeding and then recovered which ensures all reactors had the same conditions, no
gradually until the next feeding. Therefore, with an OLR statistical difference was observed among all reactors.
increase, daily batch-fed reactors experienced wider On the other hand, the continuously fed reactors showed
daily pH extremes. This is probably the reason why the lowest gas production, while the non-mixed reactors
the continuously fed reactors failed later than the batch- showed the highest gas production at both temperatures
fed reactors. The anomaly of the superior performance implying the importance of closer microbial consortia
of the MNMR during the start-up period and the failure proximity (Fig. 10).
of MNMR at the lowest OLR is probably due to the
batch feeding induced pH drops and/or the lack of 3.4. A kinetic model for describing propionate
inorganic nutrient supplementation because only the degradation by syntrophic reaction
MNMR was not supplemented with nutrients (Figs. 3
and 8). All other reactors were supplemented with McCarty and Smith [10] have graphically illustrated
nutrients since Day 69. The MNMR was never the thermodynamics of propionate and H2 metabolism
supplemented, nor appeared to need supplementation as a function of the H2 concentrations. Propionate
with nutrients up to Day 350, when propionate started conversion to acetate and H2 is thermodynamically
to accumulate. This can also be explained by the failure possible only at H2 concentrations below 104 atm. H2
of TNMR at the highest OLR because TNMR having conversion to methane is possible only at H2 concentra-
the same reactor conguration as MNMR was supple- tions above 106 atm. Therefore, H2 concentrations in
mented by inorganic nutrients early in the study (Day the bulk liquid must be in the range of 104106 atm.
69). Therefore, inorganic nutrients appear to be more The extent of propionate degradation is a function of
important at high OLR. propionate concentration and can be modeled by the
Fig. 9 shows the long-term monitoring of all reactors Monod equation
for propionate. After the start-up period, all thermo-
kX Pr
philic reactors except TNMR showed an increase in dPr f1 Pr dt; 1
KS Pr
propionate concentrations as the OLR increased. On the
other hand, all mesophilic reactors except MTP2 showed where Pr is the propionate concentration in bulk liquid
little increase of propionate concentrations. All reactors (mg/L COD), k the maximum specic propionate
showed no other VFA concentration accumulation until removal rate (day1), X the biomass (mg/L VSS), KS
reactor failure. The propionate concentration of TNMR the half-velocity substrate concentration (mg/L COD).
did not show up until reactor failure except for a short McCarty and Smith [10] also showed that higher
time (from Days 290 to 330). Therefore, these results hydrogen partial pressures tend to supply more energy
imply that the problem of the poor efuent quality in to the hydrogen-consuming species, whereas lower
thermophilic processes can be solved by non-mixing (i.e. partial pressures result in more energy going to the
closer microbial consortia proximity). hydrogen-producing bacteria. Therefore, the extent of
Calculating average VS destruction percentage and propionate degradation is hypothesized to be a function
cumulative gas production from Day 239 (after 3 HRT of negative free energy change. If it is assumed that the
M. Kim et al. / Water Research 36 (2002) 43694385 4381

Fig. 9. Long-term monitoring of all reactors for propionate.

relationship between the extent of propionate degrada- If it is assumed that dPr f1 Prf2 DGPr ;
tion and negative free energy change is linear in the  
kX Pr 
reaction of propionate conversion to acetate and dPr f1 f2 dt A 1:25 logpH2  5 : 4
KS Pr
hydrogen (Pr-Ac+H2), the following equation can
be obtained: Assuming A 1 kcal1, k 6 day1, KS 200 mg/L,
and X 1000 mg/L, the extent of propionate degrada-
dPr f2 DGPr ADGPr ; 2 tion can be expressed as
 
where A is the propionate removal coefcient (kcal1). 6000 Pr 
dPr dt 1:25 logpH2  5 ; 5
From their illustrations [10], DGPr is a function of 200 Pr
pH2 and can be expressed as
where log (pH2)o4.
DGPr 1:25 logpH2  5; 3 Using Excel spreadsheet software, the extent of
propionate degradation is severely retarded with an
where pH2=H2 partial pressure in bulk liquid. increase of H2 partial pressure, as shown in Fig. 11.
4382 M. Kim et al. / Water Research 36 (2002) 43694385

Fig. 10. Average VS destruction and cumulative gas production (from Days 239 to 345, error bars represent 2 standard deviations).

substrate concentration (mg/L COD), where


6olog(pH2).
Assuming B 1 kcal1, k 6 day1, KS 0:0005
mg/L, X 1000 mg/L, the extent of H2 degradation
can be expressed as
 
6000H2 
dH2 dt 1:33 logpH2 8 : 7
0:0005 H2
Using Excel spreadsheet software, Fig. 12 shows the
extent of H2 degradation with H2 partial pressure.
At steady state (1 mol propionate (=112 g COD)
produces 3 mol hydrogen (=16 g COD)),
112
dPr dH2 2:3 dH2 ; 8
16  3
where propionate and H2 are in mg/L as COD.
Fig. 11. Effect of H2 partial pressure on the extent of The relationship between H2 partial pressure (pH2)
propionate degradation (for the interval of 1 day). and H2 molar concentration is given by Henrys law,
which at 351C is as follows: [H2](g)(atm)/[H2](aq)(mol/
Likewise, the extent of H2 degradation can be L)=1330 atm L/mol. For pH2=4  105 atm, the corre-
hypothesized by the Monod equation and McCarty sponding H2 solution concentration would be 3  105
and Smith [10]: mol/m3. Then, 3  105 mol/m3=3  105  16 g COD/
  L=4.8  104 mg/L as COD. Therefore, pH2=0.083H 2.
kX H2 
dH2 dt B 1:33 logpH2 8 ; 6 If the term of H2 is replaced by the term of C1
K S H2
(=corresponding H2 solution concentration of pH2 at
where B is the hydrogen removal coefcient (kcal1), k the surface of H2 producer, pH2=0.083 C1 mg/L as
the maximum specic hydrogen removal rate (day1), COD) in Eq. (5) and C2 (=corresponding H2 solution
H2 the H2 concentration in bulk liquid (mg/L COD), X concentration of pH2 at the surface of H2 consumer,
the biomass (mg/L VSS) and KS the half-velocity pH2=0.083 C2 mg/L as COD) in Eq. (7), from Eqs. (5)
M. Kim et al. / Water Research 36 (2002) 43694385 4383

Fig. 12. Effect of H2 partial pressure on the extent of H2


degradation (for the interval of 1 day).

and (7)(8), the following equation can be obtained:


 
6000Pr
dtf1:25 log0:083C1  5g
200 Pr
 
6000C2
2:3 dtf1:33 log0:083C2 8g: 9
0:0005 C2
McCarty and Smith [10] calculated the distance between Fig. 13. Effect of the distance between two syntrophic bacteria
two syntrophic bacteria as the OLR varies at steady and OLR on propionate removal.
state, which is shown in the following equation:
 
2D C1  C2 1=2
DZ ; 10
Rt C1 C2
where DZ is the distance between two syntrophic
bacteria (mm), C1 the H2 concentration at the surface
of propionate oxidizer (mg/L COD), C2 the H2
concentration at the surface of H2 consumer (mg/L
COD), Rt the turnover rate per second and D the
molecular diffusivity of H2 in water=5.85  105 cm2/s,
Rt can be calculated as
Rt OLR=H2 ; 11
where OLR is the organic loading rate as H2 generated
(H2 g COD/L day) and H2 the H2 concentration in bulk
liquid (H2 mg/L COD). Fig. 14. Comparison of experimental data and model predic-
At constant pH2=4  105 atm (corresponding H2 tion for propionate concentration.
concentration=6  105 mg/L COD), Rt (s1)=193
OLR and from Eqs. (10) and (11), the following As shown in Fig. 13, propionate concentration
equation can be obtained: increases rapidly at some point of OLR, which results
C2 OLR DZ2 61 in reactor failure, and the OLR at this point increases as
C1 : 12 DZ decreases. If DZ of MB is 2 mm and DZ of TNMR is
61  OLR DZ2
1.4 mm, Fig. 14 shows that this model reasonably
From Eqs. (9) and (12), the following equation can be
matches the experimental data.
obtained:

200C2 f1:33 log0:083C2 8g


Pr     : 13
C2 OLR DZ 2 61
0:43 1:25 log 0:083  5 0:0005 C 2  C 2 f 1:33 log0:083C2 8g
61  OLR DZ2

When C2 0:0001 mg/L, using Excel spreadsheet Fig. 15 also shows a signicant effect of the distance
software, Fig. 13 shows the propionate concentration, of two syntrophic microorganisms on the extent of
which is a function of OLR and DZ: propionate and H2 degradation at steady state. From
4384 M. Kim et al. / Water Research 36 (2002) 43694385

Fig. 15. Effect of the distance of syntrophic microorganisms on the extent of propionate and H2 degradation.

Eqs. (5), (7) and (12), the extent of propionate and H2 * The comparative results of the reactor perfor-
degradation vs. H2 partial pressure based on the H2 mance at steady state clearly showed the superior
concentration on the surface of the H2 producer can be performance of TNMR with respect to lower
plotted. At steady state, the extent of propionate and H2 VFA, higher gas production and VS removal
degradation should be equal, which is shown by three conrming that microbial consortia proximity can
intersections in Fig. 15. As DZ increases, the extent of solve the poor efuent quality problem of thermo-
degradation decreases, which indicates the important philic.
role of close microbial consortia proximity. * During the long-term operation with OLR increases,
the continuously fed reactors failed at the highest
OLR due to the steady pH in the reactor. The
4. Conclusions thermophilic non-mixed reactor also failed at the
highest OLR showing much better performance than
The overall conclusions from the results presented in the continuously fed reactors until reactor failure. On
this study can be summarized as: the other hand, the mesophilic non-mixed reactor,
which was never supplemented by inorganic nutri-
* During the start-up period, stable pH, lower VFA ents, failed earlier than the continuously fed reactors
concentrations, higher gas production, and stable VS even though it showed much better performance than
removal of non-mixed reactors at both temperatures the continuously fed reactors prior to reactor failure.
clearly showed efcient performance between acid This result conrms the important role of both close
producers and consumers. This implies the impor- microbial consortia proximity and nutrients supple-
tance of close microbial consortia proximity to mentation.
enhance microbial scavenging of dissolved H2 con- * Two-phase digestion at both temperatures did not
centrations to lower levels, improving the thermo- show any signicant benet. However, thermophilic
dynamic advantage of the process at both two-phase showed slightly better performance than
temperatures. Therefore, thermophilic disadvantages mesophilic during both the start-up and the long-
can be solved by the efcient use of close microbial term periods.
consortia proximity. * The results for the same conditions during OLR
* Supplementing inorganic nutrients signicantly re- increase also clearly showed that the non-mixed
duced all VFA concentrations dramatically, except reactor conguration at both temperatures removed
for propionate. On the other hand, in the case of the VFAs more efciently and produced more gas than
non-mixed reactor conguration, propionate was other reactor congurations conrming efcient
also considerably reduced. Therefore, it turned out conversion of solubilized compounds to CH4 with
that nutrients bioavailability played an important propionate below detection concentration which is
role in anaerobic digestion at both temperatures. quite signicant.
* Propionate was efciently reduced by the non-mixed * The proposed model for the effect of the distance
reactor conguration which enhanced close microbial between two syntrophic bacteria reasonably matched
consortia proximity. the experimental data in this study.
M. Kim et al. / Water Research 36 (2002) 43694385 4385

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