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Influence of organic loading rate on methane

production in a CSTR from physicochemical sludge
generated in a poultry slaughterhouse
a a b
Luis A. López-Escobar , Sergio Martínez-Hernández , Grisel Corte-Cano & Juan M. Méndez-
b
Contreras
a
Instituto de Biotecnología y Ecología Aplicada (INBIOTECA), Universidad Veracruzana,
Xalapa, Veracruz, México
b
División de Estudios de Posgrado e Investigación, Instituto Tecnológico de Orizaba, Orizaba,
Veracruz, México
Published online: 16 Oct 2014.

To cite this article: Luis A. López-Escobar, Sergio Martínez-Hernández, Grisel Corte-Cano & Juan M. Méndez-Contreras
(2014) Influence of organic loading rate on methane production in a CSTR from physicochemical sludge generated in a
poultry slaughterhouse, Journal of Environmental Science and Health, Part A: Toxic/Hazardous Substances and Environmental
Engineering, 49:14, 1710-1717, DOI: 10.1080/10934529.2014.951258

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 Journal of Environmental Science and Health, Part A (2014) 49, 1710–1717
Copyright © Taylor & Francis Group, LLC
ISSN: 1093-4529 (Print); 1532-4117 (Online)
DOI: 10.1080/10934529.2014.951258

Influence of organic loading rate on methane production in a

CSTR from physicochemical sludge generated in a poultry
slaughterhouse

LUIS A. LOPEZ-ESCOBAR 1 
, SERGIO MARTINEZ-HERNANDEZ1
, GRISEL CORTE-CANO2

and JUAN M. MENDEZ-CONTRERAS 2

1
Instituto de Biotecnología y Ecología Aplicada (INBIOTECA), Universidad Veracruzana, Xalapa, Veracruz, M exico
2
Division de Estudios de Posgrado e Investigaci
on, Instituto Tecnol
ogico de Orizaba, Orizaba, Veracruz, M
exico
Downloaded by [Sergio Martinez] at 15:42 20 October 2014

The influence of the increase of the organic loading rate (OLR) on methane production in a continuous stirred-tank reactor (CSTR)
from physicochemical sludge generated in a poultry slaughterhouse was evaluated. Total solid (TS) to obtain OLR of 1, 5, 10 and
15 g VS L¡1 day¡1, with hydraulic retention times of 29, 6, 6 and 4, respectively, were conditioned. The results showed a decrease in
pH levels and an increase in the theoretical volatile fatty acids (VFA). While the yield of methane production decreased from 0.48 to
0.10 LCH4/g VSremoved, respectively, the OLR-10 managed on average 38% removal of volatile solids (VS) and a yield biogas
production of 0.81 Lbiogas g¡1 VSremoved and 1.35 L day¡1. This suggests that the OLR increases in an anaerobic system from
physicochemical sludge only inhibits the methanogenic metabolism, because there is still substrate consumption and biogas
production.
Keywords: Anaerobic digestion, CSTR, inhibition, methane production, OLR, pH, physicochemical sludge.

Introduction loading rate of 1 kgVS m¡3 reactor d¡1 and a hydraulic

Anaerobic digestion is a bioprocess that converts organic
matter into energy in biogas form, using microbial consor-
tia (hydrolytic, acidogenic and methanogenic) that involve
a series of biochemical reactions.[1] At the same time, they
can produce stabilized organic materials that can be used
as fertilizers or soil improvers. Actually, such mechanisms
that inhibit the methanogenic metabolism from anaerobic
digestion, with the aim of producing hydrogen have been
explored.[2–5] In this sense, there have been studies to inves-
tigate the effect of environmental parameters such as pH,
temperature and the organic loading rate (OLR), defined
as the mass of substrate added per day per reactor vol-
ume,[6] on inhibition of the methanogenesis process. Gen-
erally, anaerobic digestion processes work with an organic
Address correspondence to Juan M. Mendez-Contreras, Divi-
si
on de Estudios de Posgrado e Investigaci on, Instituto Tec-
nologico de Orizaba, Av. Tecnol ogico No. 852, Col. Emiliano
Zapata, C.P. 94320, Orizaba, Veracruz, Mexico; E-mail:
jmmendez@itorizaba.edu.mx
Received May 3, 2014.
Color versions of one or more of the figures in this article can be
found online at www.tandfonline.com/lesa.
retention time (HRT) in a range of 20–40 days.[7]
On the other hand, when the OLR is increased, the
level of pH decreases (<6.5) and the anaerobic system is
acidified, due to an increase in the concentration of vola-
tile fatty acids (VFA).[8–10] This suggests that the metha-
nogenic consortia metabolism is inhibited, because the
optimal pH for methanogens is around 7, without caus-
ing hydrogen consumption.[11] A similar work showed
that the biogas yield and methane percentage decreased
(0.85–0.65 L g¡1, and 58–50%, respectively) when the
OLR increased (0.8 to 3.4 g L¡1 d¡1) in a thermophilic
continuously stirred-tank reactor (CSTR).[12] It has also
been shown that an increase in the solids retention time
(SRT) (3–20 days) decreases the OLR, increasing the
methane percentage (4 to 55%) and decreasing the hydro-
gen percentage (68 to 4%) in a CSTR fed with synthetic
wastewater containing glucose.[13] It has been observed
that the methane yields in anaerobic digestion and varies
according to the OLR of flow rate of the anaerobic
system.[14]
Some waste biomass (barley, potato, cow dung and
organic fraction of municipal solid waste (OFMSW)) have
been used for the production of methane.[7,12,15,16] In this
regard, physicochemical sludges generated in the treat-
ment of wastewater from a poultry slaughterhouse are a
 Organic loading rate on CH4 production from physicochemical sludge
potential resource for the methane production, because it
is a waste with high concentration of organic matter,
abundance and low cost feedstock.[17] However, few
researchers have focused on studying this waste and
anaerobic digestion. For example, it is known that anaer-
obic digestion was used to stabilize the physicochemical
sludge generated in the treatment of wastewater from a
poultry slaughterhouse, with an OLR of 1.76 KgVS m¡3
d¡1,[18] while Ruiz-Espinoza et al.[17] tested OLRs of 1, 2
and 3 KgVS m¡3 d¡1, respectively, watching these load
levels achieve a biogas production on average of 2.14, 3.6
and 5.2 L d¡1, respectively, both studies conducted in a
CSTR.
Nevertheless, there is no information available on the
behavior of pH, VFA, biogas yield and methane composi-
tion, with OLRs <3 KgVS m¡3 d¡1, with physicochemical
sludge. As such, determination of the influence of OLR
Downloaded by [Sergio Martinez] at 15:42 20 October 2014
increase in the methane production from physicochemical
sludge still must be reported. At the same time, it is impor-
tant to know if the OLR can be used as a variable that
allows stable operation of an anaerobic process regardless
of the scale of a CSTR. In this sense, the aim of this
research was focused on the study of methanogenesis inhi-
bition, when the OLR increases from physicochemical
sludge generated in the treatment of wastewater from a
poultry slaughterhouse, evaluating the pH, total theoreti-
cal VFAs, biogas composition and methane yield as
response variables. In addition the OLR was calculated in
terms of the concentration of total solids (TS), not by the
flow rate.
Materials and methods
Inoculum and substrate
The organic substrate chosen was sludge generated at
a wastewater treatment plant from a poultry slaugh-
terhouse, located in Veracruz State, Mexico. In the
plant, the treatment used is a coagulation-flocculation
process, also called advanced primary treatment
(APT), classifying the sludge generated as physico-
chemical. Its characteristics are shown in Table 1.
Two continuous-flow stirred tank reactor (CSTR-1 and
CSTR-2, respectively) with built-in fiberglass (Fig. 1),
were operated and monitored for more than one year, with
an OLR of 1 gSV L¡1 d¡1 in a semicontinuous and a
Table 1. Characterization of poultry slaughterhouse sludge.
Parameters Units Value
pH 6.7 § 0.2
TS % m v¡1 2.5 § 0.3
VS % m v¡1 2.1 § 0.4
N-NH3 g N- NH4 kg¡1 TS 4.49 § 1.2
(Data are given as mean § SD, n D 5).
1711
hydraulic retention time (HRT) of 25–29 days. Then
CSTR-1 and CSTR-2 with two useful volume-scales (6 L
and 1.2 m3, respectively), were compared with the experi-
mental units (lab-scale reactors) when they were imple-
mented and operated at OLR 1 gVS L¡1 d¡1, during the
experimental strategy of this study. To set up the experi-
mental units, methanogenic seed sludge (inoculum)
obtained from CSTR-1 was used, with a pH of 7.23 § 0.2,
total theoretical VFAs of 1.4 § 0.8, total solids (TS) and
solid volatile (VS) of 15 and 9 g L¡1 respectively, and an
ammonia nitrogen concentration of 7.85 § 0.97 g N -
NH4 kg¡1.
Experimental unit
To test the different OLRs, lab-scale reactors in semi-con-
tinuous flow (glass bottles of 500 mL total volume) were
inoculated with a useful volume of 350 mL of seed sludge
from CSTR-1. The reactors contained ports for biogas
sampling, feeding and unloading. The reactors were main-
tained to an orbital mixer value of 110 rpm and a tempera-
ture of 28 § 2
C. The amount of biogas produced was
recorded daily using the water displacement method.
Experimental setup
The experiments were divided into two periods: the first,
stabilization of the anaerobic system and the second, an
increase in the OLR. All experiments units were operated
to an -OLR of 1 gVS L¡1 d¡1, for a period of 25 days until
its stabilization. Subsequently, the experimentation time to
evaluate treatments (increase OLR) was 36 days (Table 2).
To evaluate the influence of OLR on the methane produc-
tion, OLRs of 1 (TC), 5 (T1), 10 (T2) and 15 (T3) gVS L¡1
d¡1, respectively, were tested, corresponding to an HRT
of 29, 6, 6 and 4 days, respectively. The OLR calculation
was made in terms of TS concentration, not of flow rate,
which was calculated as Q D A £ V/C, where Q (L day¡1)
is the flow rate, A is the OLR (gVS L¡1 d¡1), V (L) is the
reactor useful volume and C (gVS L¡1) is the feed
concentration.
Then raw physicochemical sludge was prepared with dif-
ferent concentrations of TS (34, 34, 70 and 70 g L¡1, respec-
tively) and the percentage of VS remained constant (88 §
0.2%). The experimental conditions are shown in Table 2.
The response variables used were pH, alkalinity, total theo-
retical VFA and methane percentage (% CH4) present in the
generated biogas. Also in order to define the OLR, which
better promotes the inhibition of the methanogenic process,
substrate removal efficiency (%VSremoved, (g substrate
removed [g substrate fed]¡1) £ 100); substrate removed
(gVS, g substrate fed- g product); yields biogas (Ybiogas,
L-biogas [g substrate removed]¡1) and methane (YCH4,
L-methane [g substrate removed]¡1) were determined.
 1712
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Fig. 1. Anaerobic Digestion System in a CSTR. a) Hydraulic diagram, b) Pilot-Set of 6 L, c) Pilot-Set of 1.2 m3.
Analytical methods
The TS, VS and alkalinity were analyzed according to
standard methods.[19] The pH was monitored using a pH
meter (Orion250a model). Total theoretical VFA concen-
trations were determined according to Jenkins et al.,[20]
using Eq. 1:
Alct ¡ AlcB
VFA D
0:83  0:85
where Alct is the total alkalinity titration to pH 4.3, AlcB is
the partial alkalinity titration to pH 5.7; 0.85 is a factor,
assuming that 85% of the volatile acids (which are mostly
acetic an propionic acids) have been titrated at pH 4.3;
finally 0.83 is a factor, also assuming that 83% of the vola-
tile acids is titrated to the total alkalinity endpoint. The
Lopez-Escobar et al.
ammonia nitrogen was analyzed according to method
4500-NH3.[19] The biogas contents, including O2, N2 and
CH4 were analyzed using a gas chromatograph (Thermo
Scientific Focus GC), equipped with a Heliflex AT-Mole
Sieve column (30 m long £ 0.53 mm diameter) and a ther-
mal conductivity detector. Temperatures of the injection
port, oven and the detector were 250, 35 and 121
C,
respectively. Helium was used as carrier gas with a flow-
rate of 60 mL min¡1.
(1)
Statistical analysis
The values presented in the tests are the average of four
replicas and were analyzed by ANOVA test with a confi-
dence level of 99.95% (P D 0.05), using Duncan’s post hoc
tests to determine a possible significant difference between
treatments.
 Organic loading rate on CH4 production from physicochemical sludge
Table 2. Operational conditions in lab-scale CSTR for evaluation of increasing OLR.
Operational time (days)
Treatment OLR (gVS L¡1 d¡1) Period-1 Period-2
TC 1 25 36
T1 5 25 36
T2 10 25 36
T3 15 25 36
Results and discussion
The startup of CSTR pilot-scale reactors as a source
of inoculum
The removal of volatile solid was evaluated for more than
one year (%VS) (Fig. 2), along with the production and
composition of biogas in both anaerobic reactors (CSTR-
Downloaded by [Sergio Martinez] at 15:42 20 October 2014
1 and CSTR-2), with an OLR 1 gVS L¡1d¡1. A 38%
removal of VS was used as the operational criterion to
meet the vector attraction reduction requirement indicated
by the US EPA.[21] Reactor CSTR-1 had an average VS
removal of 67%, whereas the CSTR-2 removal was 53%
(Fig. 2), keeping the two systems above 38%. The average
values of biogas production in the CSTR-1 and CSTR-2
reactors were of 5.51 and 575 LBiogas d¡1 with methane
yields of 0.52 and 0.42 LCH4 g¡1SVremoved, respectively.
The yields found in these assays (Table 3) coincide
within the range of values reported for conventional anaer-
obic digesters.[22] Also, these values were used to compare
pilot-scale anaerobic systems (CSTR-1 and CSTR-2) oper-
ated with OLR-1, with lab-scale anaerobic systems with
OLR-1 (TC) (Table 3 and Fig. 2). The behavior in meth-
ane yields and removal of VS values was similar between
lab-scale and pilot-scale anaerobic systems when both
were operated with OLR of 1 gVS L¡1d¡1. This suggests
that the OLR can be used as a variable that allows the sta-
ble operation of the anaerobic system in a CSTR,
Fig. 2. %VS removal efficiency with OLR of 1 gVS L¡1 d¡1. pH

CSTR-1 ( ), pH CSTR-2 (&), CSTR-1 (~), CSTR-2 (&),
¡38% SV-US-EPA (& & &).
1713
HRT (days) pH TS (g L¡1) VS (g L¡1) Temp. (
C)
29 7.73 34 30 27 § 2
6 7.81 34 30 27 § 2
6 7.66 70 63 27 § 2
4 7.76 70 63 27 § 2
regardless of the scale of work, using physicochemical
sludge as a substrate.
Effect of the OLR in the behavior of the pH and VFAs
After 36 days of operation during increasing the OLR in
semi-continuous mode, the results show that levels of pH
and VFA up to 5.7 § 0.01 and 5.53 § 0.14 g L¡1, respec-
tively were obtained (Table 4). In terms of pH, T1 had no
significant difference with regards to TC, both maintained
values close to neutrality. The final concentration of VFA
in T1 was 3.99 § 0.21 g L¡1; some studies suggest that this
level does not inhibit mesophilic methane production.[9–23]
The increment of OLR in T2 and T3 is related to the
decrease in pH levels (6 and 5.7, respectively) and the final
concentrations of VFA (5.06 and 5.53 g L¡1, respectively).
The maximum decrease in pH achieved in T3 was pre-
sented on day 52 of experimental operation and remained
in the range of 5.7 § 0.01 until the end of the experiment
(Fig. 3). These results showed a significant effect of the
OLR (Fo D 157.04) over the pH variable.
The results shown in this study are similar to those pre-
sented by Blume et al.,[24] who obtained a decrease in pH
values of 7.5 to 5.5 when propionic and acetic acid concen-
trations increased from 1.2 and 0.3 g L¡1 to 6.1 and 8 g
L¡1, respectively, by increasing the OLR with maize silage
in a lab-scale CSTR. On the other hand, it has been
reported that higher concentrations of VFAs in anaerobic
systems have pH levels of 5 to 6. It has also been reported
that through the concentration of VFA, the possible inhi-
bition of the methanogenic metabolism can be pre-
dicted.[23] Further, it is noted that inhibition of this process
occurs when the pH levels are between 5.5 and 6.[14] This is
because the methanogenic consortia grow in a relatively
narrow range of pH (6.5–7.8). However, there are studies
that mention that some acidophilic species such as Metha-
nobacterium espanolae grow at a pH between 5.6 and 6.2,
but are unable to produce methane at pH 4.7.[25] These
results suggest that T1 could not inhibit the metabolism of
an anaerobic methanogenic system operated at semi-con-
tinuous conditions and fed with physicochemical sludge.
Duncan’s post hoc test showed that T2 exhibited a signifi-
cant difference regarding TC and T1 according to pH lev-
els achieved (pH D 6.05). This could indicate that the
buffer capacity of the system fails to control the concentra-
tion of VFAs (5.06 g L¡1).
 1714 Lopez-Escobar et al.

Table 3. Summary of operational performances of pilot-scale CSTR-1 and CSTR-2 reactors.

Average

OLR Biogas Biogas yield Methane yield

Reactor HRT (d) pH (gVS L¡1 d¡1) VS removed (%) production (L d¡1) (L g¡1VSremoved) (L g¡1VSremoved)
CSTR-1 25–29 7.16 1 67 § 4 5.51 § 0.26 0.86 § 0.1 0.52 § 0.06
CSTR-2 25–29 7.03 1 53 § 4 575 § 13 0.71 § 0.07 0.42 § 0.04
(Data are given as mean § SD, n D 85).

Effect of OLR on the biogas and CH4 production (Fig. 4a) that biogas production decreased in the T1, T2
and T3 as operating days passed. This behavior is observed
During the stabilization phase the four treatments showed
in other studies, indicating that the growth rate of the
an average 0.15 L d¡1 of biogas and a methane concentra-
methanogens consortia (Archeas) is less than the acidogenic
tion of 60%. The increment of OLR occurred from day 25
(bacteria) (0.0167 and 0.083 h¡1, respectively).[14]
of experimental operation in T1, T2 and T3. This coin-
In this regard, the values shown in Table 3 indicate sig-
cided with an increase in the biogas volumes reported at
Downloaded by [Sergio Martinez] at 15:42 20 October 2014

nificant differences of the OLR factor on biogas yields (Fo

the end of the operation, which were of 0.38 (T1), 1.35
D 46.65) and methane yields (Fo D 248.69). This indicates
(T2) and 0.86 (T3) L d¡1, respectively, (Fig. 4a), compared
that the methanogenic phase is not capable of withstand-
to the treatment control which was 0.19 L d¡1, while the
ing high OLR. By contrast, the acidogenic anaerobic sys-
final average of methane concentration was 64 (TC), 60
tem studies indicate whether they are able to withstand
(T1), 31 (T2), and 20 (T3)%, respectively (Fig. 4b). At the
high OLR.[8] This can be supported with the values shown
end of the experiments, the percentage of CH4 in T1
in Table 2, which shows that the TRH in the TC was
showed no significant difference with regards to the TC;
29 days, and 6, 6 and 4 days for T1, T2 and T3,
however, with T2 and T3 there were significant differences
respectively.
(Fo D 2188.15), showing decreases from 57 to 31% in T2
The behavior of our study is comparable with the
and from 61 to 20.15% in T3 (Fig. 4b).
results found by Lee et al.,[8] in a methanogenic reac-
These results indicate that the OLR factor had a signifi-
tor simultaneously fed with a residual substrate from
cant effect on the methane production of the methanogenic
an acidogenic fermenter, at the same time fed with
metabolism, using physicochemical sludge as substrate.
food waste in the form of OFMSW. In this study a
These results are consistent with previous studies, which
stable pH was maintained in the range of 7.4–7.9,
suggest that an overload (maize silage  4.1 g dry organic
between the OLRs of 4.16 and 8.4 gCOD L¡1d¡1,
matter L¡1 d¡1) in a conventional anaerobic digestion sys-
operating without pH regulation using an alkali chemi-
tem, used in a CSRT, caused process instability and the dis-
cal buffer. Afterwards, with OLR of 11.8 gCOD
appearance of aceticlastic Methanosaetaceae, which
L¡1d¡1 acidification in methane, fermentation abruptly
predominated at lower OLRs.[24] Such would be related to
occurred, and the pH dropped from 7.42 to 6.4 due to
an increment in the concentration of VFAs produced in the
the accumulation of VFAs after 90 days of semi-con-
acidogenic phase.[9,24] This behavior was also found in our
tinuous operation, indicating that the decrease in pH
experiments (Figs. 4a and 4b). At the beginning of the over-
is attributed to the accumulation of acetate and propi-
load a correlation was observed between the increase of the
onate, resulting in the reduction of methanogenic activ-
OLR and the biogas production (R2 D 0.99). This could be
ity. This coincided with a decrease in the percentage of
attributed to a synergy between the microbial communities
CH4 (80–20%).
in the anaerobic system.[13] However, it can be observed

Table 4. Final values obtained at conclusion of experiments.

Consumption Yield

Ta OLR b pH Alkalinity c VFAs d Efficiency e VSremoved f YBiogsg YCH4h

TC 1 7.41 4.03 2.57 59.96 0.25 0.75 0.48
T1 5 7.28 3.93 3.99 37.07 0.81 0.47 0.29
T2 10 6.05 3.86 5.06 41.79 1.67 0.81 0.25
T3 15 5.70 3.90 5.53 29.23 1.76 0.49 0.10
a D treatment; b D gVS [L d]¡1; c D g [L as CaCO3] ¡1; d D g [L as HAc.] ¡1; e D %VSremoved; f D grams;
g D LBiogas [g VS removed]¡1; h D Lmethane [g substrate removed]¡1. (Data are given as mean § SD, n D 4).
 Organic loading rate on CH4 production from physicochemical sludge
Downloaded by [Sergio Martinez] at 15:42 20 October 2014

Fig. 3. OLR effect on a) pH and b) VFAs. TC ( ), T1 (&), T2
(~), T3 (!).
pH and %VSremoved variation
The final pH and its average variation (DpH D pHinicial –
pHfinal) in the treatments are observed in Figure 5. The
final pH was conditioned by the acid concentration. The
final pH in the control treatment was 7.4 § 0.02, while for
treatments 1, 2 and 3 it was 7.2 § 0.71, 6.0 § 0.12 and 5.7
§ 0.01. The largest decrease in DpH in T2 (1.61) was
observed, reaching a pH of 6.05 (Fig. 5). In T3, a signifi-
cant acidification was not observed, which may be attrib-
uted to the acidity caused by the weak and organic nature
of OLRs in acidogenic anaerobic systems, which reached a
threshold between 5 and 5.5. On the other hand, the final
values of organic material removal (% VSremoved), as well
as the average removed variation (D%VSremoved D
%VSremoved-initial –%VSremoved-final) are shown in Figure 5b.
It was found that the VS removal efficiency decreased as
the OLR increased.
However, the VS removal efficiency was less than 38%
when the OLR was more than 15 gVS L¡1 d¡1. The delta
values in T1 and T2 were lower (26.75 and 26.67) com-
pared with T3. These results indicate that for greater delta
value, the VS removal efficiency will be low. A similar
study showed similar behavior in a methanogenic reactor
1715
Fig. 4. Behavior of a) biogas, b) CH4-concentration, during the

increase of OLR. Control ( ), T1 (&), T2 (~), T3 (!).
operated in a semi-continuous manner, fed by pre-digested
sludge in an acidogenic fermenter with OLR of 4.16, 8.4,
and 11.8 gCOD L¡1 d¡1 and removals of 88.1, 86.3 and
83.3%, respectively.[8] This could happen due to the mass
transfer between anaerobic microorganisms and the sub-
strate, which is affected by the high concentration of total
solids, causing a decrement in the efficiency of organic
material removal.[26,27]
Other studies suggest that the acidogenic consortia (bac-
teriae) are responsible for keeping the efficiency of elimina-
tion of VS because the rate of growth is higher in regard to
the methanogenic consortia (Archeas),[14] as in the case of
T2 in our study, with final pH values of 6. Therefore, is
possible that the methanogenic consortia could be kept at
a minimum concentration but sufficient to degrade part of
the metabolites produced by the acidogenic consortium.
This maintained the anaerobic metabolism of the 38% VS
removal criterion for the vector attraction reduction
requirements indicated by the US EPA.[21]
The results found in this study using physicochemical
sludge as organic substrate in CSTR reactors indicated
that increased OLR had a negative influence on produc-
tion and yields of methane. However, the biogas produc-
tion and the removal of organic matter expressed as %SV,
 1716 Lopez-Escobar et al.

substrate and mixed cultures as inoculum (Table 5). For

example, inhibition of methanogenesis has happened by
controlling pH at acidic levels (5.5, 5.9 and 5.0) with the help
of external acidic agents and as organic substrate cheese
whey (CW), cheese whey powder (CWP), and organic frac-
tion of municipal solid waste (OFMSW).[28–30] Also, pre-
treatment of a methanogenic inoculum achieved the
inhibition of methane production in anaerobic digesters,
maintaining acidic pH levels (5.5 and 5.2–5.8), using
OFMSW and food waste as substrate. [8,9]

Conclusions

Our results suggest that the OLR can be used as a variable

allowing stable operation of anaerobic digestion, regardless
of the scale of work. Also, the OLR promotes the inhibitory
Downloaded by [Sergio Martinez] at 15:42 20 October 2014

mechanism of the methanogenic activity in the CSTR reac-

Fig. 5. Effect of the OLR on final values of the pH and % VS

removed. a) pH ( ), DpH (&), b)%VSremoved. (~), D%VSremoved.
(!).
was not inhibited in its entirety. These results are of great
interest given the nature of substrate used (physicochemi-
cal sludge) considered complex because its high concentra-
tion of organic material (85% VS) and the presence of salts
of aluminum and iron.[22] It is important to mention that
according to our knowledge, there are no studies in the lit-
erature using this type of substrate.
However, some studies with similar results could be men-
tioned from experiments operating in continuous and semi-
continuous anaerobic systems, using residual biomass as
tors, using physicochemical sludge as substrate produced
during the wastewater treatment from a poultry plant pro-
cessor. In addition, the calculation of the OLR in terms of
TS allowed evaluation of flow rates with high organic mat-
ter concentrations (10 gVS L¡1 d¡1) per unit of volume
and low HRT (< 6 days). At these conditions, low pH lev-
els in T2 and T3 (6 and 5.7, respectively) as well as a
decrease in methane concentration up to 20% was observed.
However, generated biogas in T2 values higher than
38% VSremoved was seen, despite having presented acido-
genic conditions. In contrast the experiment tests TC and
T1 kept their pH levels neutral (7) and presented a con-
stant methane concentration (62%). Future studies could
use this operational strategy for promotion of conditions
that allow the production of biohydrogen, without using
external acidic agents that will influence the fall of pH in
anaerobic reactors (CSTR).
Funding
L
opez-Escobar acknowledges support from a PhD schol-
arship from the Council of Science and Technology of
Mexico (CONACyT) with CVU (Grant holder) 168870/
168870. In addition, he acknowledges the support from

Table 5. Data reported in literature for anaerobic digestion operated at low pH and with other waste biomass.
Temp HRT CH4
Substrate Inoculum (
C) (Days) OLR pH (L g¡1 d¡1) References
* y
OFMSW Anaerobic seed sludge 55 1.3 38.4 5.5 ND [29]
* z
CWP Anaerobic seed sludge 37 6 to 3.5 95 to 163 5.9 ND [28]
* z
CW Composting with waste food 30 1 20 and 30 5 ND [27]
y
Waste food **
Anaerobic granular sludge 37 4 to 10 15.10 to 37.75 5.2 – 5.8 ND [12]
** z
OFMSW Anaerobic seed sludge 55 3.6 to 1.3 19.5 to 58.5 5.5 3.1–7.67 [11]
§ y
Physicochemical sludge Anaerobic seed sludge 26 6 to 4 5 to 15 5.7 0.45 to 0.1 This study
§
Untreated, *pH control with external acidic agents, **inoculum-pretreated, ygVS L¡1 d¡1, zgCOD L¡1d¡1, ND D No Detected, OFMSW D
Organic fraction municipal solid waste, CWP D cheese whey powder, CW D cheese whey.
 Organic loading rate on CH4 production from physicochemical sludge
the infrastructure of the Environmental Laboratories of
the postgraduate of the Instituto Tecnologico de Orizaba.
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