Bioresource Technology 96 (2005) 1153–1159

Respirometric assays at fixed and process temperatures to monitor

composting process
Raquel Barrena Gómez, Felı́citas Vázquez Lima *, Ma.Angels Gordillo Bolasell,
Teresa Gea, Antoni Sánchez Ferrer
Escola Universitaria Politécnica del Medi Ambient, Universitat Autónoma de Barcelona, Rambla Pompeu Fabra 1,
08100 Mollet del Vallès (Barcelona), Spain

Received 1 March 2004; received in revised form 22 September 2004; accepted 30 September 2004
Available online 2 December 2004

Abstract

A static respirometer was built to determine the respiration index (RI) of composting samples. Respiration indices of different
sludges were determined at 37 °C (RI37) and at the in situ temperature of the composter at sampling (RIT). Results indicated that
both indices correlated well with temperature evolution in the composter. RIT were more representative of the metabolic activity in
the reactor and more sensitive to temperature and composition variations of the composting material, but could not indicate the
stability of the material at later stages of the process. Moreover, significance of RI units was shown in the composting of a highly
compostable residue. According to the stability limits suggested in the literature, initial RI expressed on dry matter (DM) basis cor-
responds to a stable material (RI < 1 mg O2 g DM
1 h
1) whereas initial RI expressed on organic matter basis (OM) corresponds to
an unstable material (RI = 2.5 mg O2 g OM
1 h
1).
Ó 2004 Elsevier Ltd. All rights reserved.

Keywords: Monitoring; Respiration index; Stability

1. Introduction bolic activity; methods such as total ATP, enzyme activ-

ity and microbial biomass measurements have been
Composting has received much attention as a poten- employed (Garcı́a et al., 1992; Tiquia and Tam, 1997;
tial technology for treating solid wastes such as munici- Garcı́a and Polo, 1999; Benı́tez et al., 1999; Tiquia
pal solid wastes, sewage sludge, yard trimmings, and et al., 2002). However, the most widely used biological
agricultural wastes (De Vault, 2004; Grigatti et al., methods are the respirometric studies which determine
2004; Brewer and Sullivan, 2003; Aziz et al., 1992). Dur- the O2 or CO2 evolution during the microbial metabo-
ing the last decades research has focused on the qualita- lism (Kuter et al., 1985; Atkinson et al., 1996; Pérez
tive evaluation of the composting process. Several et al., 1999; Larsen and McCartney, 2000; Lasaridi
indicator variables have been proposed for monitoring et al., 2000). The basis of these methods is that immature
the process and evaluating the stability of the final prod- compost has a strong demand for O2 and high CO2 pro-
uct. Biological and biochemical indices are particularly duction rates due to intense development of microorgan-
useful since they relate composting processes to meta- isms as a consequence of the easily biodegradable
compounds in the raw material. Conversely, at late com-
posting stages both processes decline as the amount of
*
Corresponding author. Fax: +34 93 579 6785. degradable organic mater decreases. However, it has
E-mail address: [email protected] (F. Vázquez Lima). been argued (Haug, 1986; Adani et al., 2001) that O2

0960-8524/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2004.09.026
1154 R.B. Gómez et al. / Bioresource Technology 96 (2005) 1153–1159

consumption is a more reliable method since oxygen tained from the de-inking process of a recycled paper
is directly responsible for the oxidation of the organic manufacturing industry.
matter. Still, O2 monitoring requires more control and Wood chips, consisting of a variable mixture of pine
therefore more sophisticated equipment and is more and beech tree wood, were obtained from a local carpen-
time consuming than the measurement of CO2 try and used as bulking agent. Sludge and wood chips
production. were mixed by hand after the screening of the bulking
Methods based on O2 consumption have been classi- agent. A semi-industrial sieve (Filtra Vibración, FT-
fied into dynamic and static protocols (Adani et al., 400) was used for the wood chips screening. Three differ-
2001). Dynamic methods are characterised by a contin- ent screens (20, 10 and 5 mm) mesh were available. A 1:1
uous oxygen supply whereas static methods can be used sludge:wood chips volumetric ratio was used in all
with solid or liquid samples. A wide range of devices has experiments since it had been previously optimised by
been developed for the measurement of respiration from Gea Leiva et al. (2003).
solid matrices. One of the most widely reported uses a
dissolved oxygen (DO) meter with a polarographic 2.2. Composter
probe measuring the decline in oxygen concentration
over a compost sample in a flask. Such tests generally A 100 l fridge was adapted to be used as a static com-
provide near-optimum conditions for microbial respira- poster for the pilot-scale experiments described in this
tion (e.g. moisture, nutrients, and oxygen supply). In al- work. It was filled either with OFMSW, the sludge:wood
most all respirometric experiments, the measurement of chips mixture or PCS.
respiration activity is carried out at standard tempera- The recipient was kept horizontal with a slight incli-
tures of about 30–37 °C (USDA, 2001; Iannotti et al., nation to allow its opening from the top and to make
1993; Lasaridi and Stentiford, 1998). It is considered the collection of possible leachates easier. A plastic mesh
that respirometric activities measured at these fixed tem- was fitted at the bottom of the recipient to support the
peratures are good indicators of the mean metabolic po- material and collect the leachates. Several holes were
tential of the compost. Nevertheless, composting is a perforated through the walls of the vessel to permit
complex process where the rate of degradation is a result the entry and exit of air, the discarding of leachates
of the metabolic activity from a mixed microbial popu- and the insertion of different probes. The air inlet was
lation that includes microorganisms with different opti- at one end of the vessel whereas its outlet was at the
mum growth temperatures. It can be considered that, other end. An air flow rate of 20 l/min was used and
although respirometric experiments performed at fixed its entry was regulated either by temperature or O2
temperatures are a useful indicator of the compost sta- control. Four Pt 100 sensors (Desin mod. SR-NOH),
bility, they do not show the actual metabolic stage of inserted at different points inside the vessel, were used
the process and, therefore, cannot be used to follow its for monitoring the temperature. Oxygen concentration
evolution. On this basis, this paper describes the exper- in interstitial air was monitored with an oxygen sensor
imental setup of a respirometer developed to determine (Sensox, Sensotran, Spain). CO2 concentration at the
the respiration index (RI) of composting samples at dif- outlet of the composter was monitored with an infrared
ferent temperatures. The final aim of this work will be to detector (Sensotran I.R., Sensotran, Spain). All sensors
have a reliable method to monitor the composting were connected to a specially-made data acquisition sys-
process. tem. Oxygen was controlled by means of a feedback
oxygen control which automatically supplied fresh air
to the reactor when oxygen concentration fell below
2. Methods 10%.

2.1. Substrate characteristics 2.3. Sampling procedure and chemical and physical
analyses
Homogenised source-separated organic fraction of
municipal solid waste (OFMSW) was obtained from Samples were collected periodically to determine
the full-scale municipal solid waste composting plant organic matter content and the respiration index. Sam-
in Granollers (Barcelona). Samples were taken directly pling procedure was as follows: first, the container was
from the composting tunnels. opened and its contents mixed with a rake; then, several
Dewatered sludge (DS), consisting of primary and samples of about the same size were collected from dif-
activated sludge, was obtained from the municipal ferent parts and different depths until a 1 l beaker was
wastewater plant in La Garriga (Barcelona). Dewatered filled with sample. The whole procedure was performed
anaerobically digested sludge (ADS) was obtained from as quickly as possible to minimise disturbances to the
the municipal wastewater treatment plant in Granollers system. Samples from OFMSW were sieved to remove
(Barcelona). Physico-chemical sludge (PCS) was ob- glass, plastics and other inerts and oversized material.
 R.B. Gómez et al. / Bioresource Technology 96 (2005) 1153–1159 1155

For respirometric experiments, humidity content was was then equilibrated with the outside pressure by
adjusted to a range of 40–50% (w/w) if required. Analyt- inserting a hypodermic needle through the stopper and
ical parameters were determined according to the stan- leaving it in place for 5 min. O2 level was then recorded
dard procedures recommended by the Test Methods every 15 s for 90 min. In all experiments three replicates
for the Examination of Composting and Compost were used. After the DO readings finished, the total vol-
(TMECC) (USDA, 2001). ume of free air space in each sample flask was deter-
mined as described in TMECC (USDA, 2001).
2.4. Respirometer
2.5. Calculation of respiration index (RI)
A static respirometer (Fig. 1) was built according to
the original model described by Iannotti et al. (1993, The respiration index (RI) of the compost sample re-
1994) and following the modifications and recommenda- ferred to total organic matter content was calculated
tions given in the TMCC (USDA, 2001). The drop of from the slope in a linear segment on the chart of O2
oxygen content in a flask containing a compost sample (%) versus time by using Eq. (1).
was monitored with a dissolved oxygen meter (Lutron V  P  32  m  60
5510, Lutron Co. Ltd., Taiwan) connected to a data RI ¼ ð1Þ
R  T  X  DM  OM
logger.
The setup included two water baths to allow the where RI, respiration index (mg O2 g OM
1 h
1); V,
carrying out of experiments at two different tempera- volume of air in flask (ml); P, atmospheric pressure at
tures simultaneously. Temperatures assayed were at a elevation of measurement (atm); m, slope of change in
fixed value of 37 °C and at the in situ temperature of percent O2 saturation per minute divided by 100; R,
the composter at the moment of sampling. Prior to the ideal gas constant (0.08206 l atm mol
1 K
1); T, temper-
assays, samples for experiments at 37 °C were incubated ature in (K); X, wet weight of compost test aliquot (g);
for 18 h at this temperature, while samples for experi- DM, fraction of total solids of a parallel sample aliquot
ments at in situ temperatures where incubated for 4 h (g DM g X
1); OM, fraction of organic matter of a par-
at such temperature. During all the incubation periods, allel sample aliquot in dry basis (g OM g DM
1).
samples were aerated with previously humidified air at
the sample temperature.
The aeration system consisted of a flask with a two- 3. Results and discussion
hole stopper and two glass delivery tubes. At the bottom
tip, the delivery tube had an aquarium air-stone to pro- 3.1. Respirometer validation
duce small air bubbles. This sparger was immersed in
water to humidify the air. The flask was always kept Amongst the modifications developed in this work it
inside the incubation bath. The delivery tube was con- is worth mentioning that air used for the aeration of the
nected to a manifold that served to deliver air to the dif- samples before the respirometric assays was saturated
ferent flasks containing the samples. The manifold was with water at the same temperature as the water bath,
constructed from plastic tubing connected by quick dis- otherwise aeration resulted in the drying of the sample.
connect fittings. Individual valves were also fitted to Moreover, the use of quick disconnect fittings and indi-
each sample tube to regulate the air flow. vidual valves allowed a better control of the aeration
Once the incubation period was finished, the DO sen- rate.
sor was assembled into the flask without removing it Once the respirometer was built, the next step was to
from the water bath. The air pressure inside the flask evaluate the error of the respirometric assays. The RI of
OFMSW samples obtained from the static composter at
different days were determined. Experiments were per-
formed in triplicate. Temperatures assayed were at a
fixed value of 37 °C (T37) and at the in situ temperature
of the composter (T) at sampling with 18 and 4 h incuba-
tion, respectively. Incubation time for the fixed temper-
ature was chosen according to the literature (Iannotti
et al., 1993; USDA, 2001). As no information was
available regarding incubation for the experiments at
the in situ temperature, different incubation times were
assayed. Results indicated (data not shown) that RI
values remained practically constant for incubation
times above 4 h. Fig. 2 illustrates a typical chart of
Fig. 1. Respirometer setup. O2% versus time and RI values for three replicates of
1156 R.B. Gómez et al. / Bioresource Technology 96 (2005) 1153–1159

25 25

20 20

15
Oxygen % 15

Oxygen %
10 10

5 5

0 0
0 20 40 60 80 100 0 20 40 60 80 100
(A) Time (min) (B) Time (min)

25

20

15
Oxygen %

10

0
0 20 40 60 80 100

(C) Time (min)

Fig. 2. Oxygen (%) versus time curves of three replicates of a OFMSW sample taken on the 4th day of composting. Respiration Indices RI37

1
1
1
1
1
obtained at 37 °C were (A) 3:90 mg O2 g O
1
2 h ; (B) 3:94 mg O2 g O2 h ; (C) 3:74 mg O2 g O2 h , respectively.

Table 1
Respiration indices and standard deviations obtained during the composting of OFMSW
Mean process temperature (°C) Time of composting (day) Temperature of the Respiration index Standard deviation
respirometric test (°C) RI (mg O2 g OM
1 h
1)
55 2 55 8.75 1.17
60 4 37 3.86 0.10
55 8 37 2.46 0.20
55 3.75 0.59
55 11 37 2.23 0.11
55 6.95 0.63
55 17 55 3.17 0.21
50 25 37 0.96 0.45
50 4.31 0.19
Results correspond to three replicates.

a same sample. As expected, oxygen % decreased with evaluated at sampling temperature were higher than
time. Table 1 shows RI values and standard deviations those for RI at 37 °C. This could be due to the fact that
obtained for the two temperature conditions assayed. the former was closer to the upper limit of the tempera-
Standard deviations from respirometric tests at the start ture compensation of the electrode (60 °C). This could
of the composting process are larger than those obtained mean that minor variations in the temperature of the
at the end probably due to the homogenisation of the water bath could lead to small oscillations in the O2
material along the process. It is also observed that at probe readings, increasing the error associated with
high temperatures (55 °C) standard deviations for RI the measurement of the slope.
 R.B. Gómez et al. / Bioresource Technology 96 (2005) 1153–1159 1157

3.2. Units of RI matter basis correlated well with the temperature profile
of the process, with high values corresponding to an
RI units are generally referred to on the basis of total unstable material during the thermophilic range and
organic matter content. However, the convenience of decreasing to 1 mg O2 g OM
1 h
1 during the mesophilic
using units based on total solids content has also been phase. These results seem to indicate that when dealing
suggested (California Compost Quality Council web with materials with high content of non-degradable
site), since the organic matter content can be very vari- material, respiration indexes should be expressed on
able depending on the heterogeneity of the material. In an organic matter basis since the high content of non-
consequence, this variability could introduce significant degradable material may lead to misleading results when
errors in the evaluation of the RI. expressing them on a dry matter basis.
Therefore, the significance of the units used for the
respiration index was evaluated during the pilot-scale 3.3. Monitoring of the composting process by
composting of physico-chemical sludge (PCS) resulting respirometric techniques
from the de-inking process of a recycled paper manufac-
turing industry. This material has a low organic matter Fig. 4 shows that composting of sludge (DS) followed
content (approximately 34% dry weight basis) which the typical evolution of this type of process. It is ob-
suggests a low compostability. However, composting served that during the first two days process temperature
at laboratory scale with no bulking and no amendment quickly rose to 65 °C followed the next day by a fast
showed good performance (data not shown). Tempera- drop to 40 °C and a less pronounced fall during the fol-
ture profile and respiration indices determined at 37 °C lowing days to temperatures as low as 20 °C. Tempera-
(RI37) obtained during the composting at pilot-scale tures then increased slowly up to values around 30 °C.
are shown in Fig. 3, where it is observed that a thermo- They were maintained in the mesophilic range until
philic range was reached within 2 days, and was main- the process was finished.
tained for more than 2 weeks. Evolution of the composting of anaerobically di-
From Fig. 3, it can also be seen that RI values calcu- gested sludge (ADS) is shown in Fig. 5. In this case, it
lated on a dry matter basis were all below can be seen that temperature rapidly reached a thermo-
1 mg O2 g DM
1 h
1 regardless of the composting stage. philic range in the first days of the process, with values
According to literature (USDA, 2001 California Com- up to 60 °C, followed by a moderate drop to around
post Quality Council web site), these values would cor- 38 °C and a subsequent quick increase up to 50 °C.
respond to a stable material. However, this is in clear Afterwards, temperature slowly decreased to the meso-
contradiction to the actual activity shown by the mate- philic range.
rial as confirmed by the temperature profile of the pro- There are two possible explanations for the tempera-
cess shown in Fig. 3. Initial RI value calculated on an ture profiles during the first stage of the process. The
organic matter basis corresponded to an unstable mate- most generally accepted is that once easily degradable
rial (RI = 2.5 mg O2 g OM
1 h
1), as suggested in the lit- materials are depleted metabolic activity slows down
erature (USDA, 2001; California Compost Quality causing a temperature drop. Microorganisms able to
Council web site). RI values calculated on an organic degrade more complex materials then develop and their

70 14
70 4.0 T
T 60 RI37 12
60 RI37 (mgO2 * gOM-1* h-1) 3.5
RIT
RI (mg O2*gOM *h )
-1
Temperature (ºC)

50 10
RI37 (mgO2 * gDM-1* h-1)
Temperature (ºC)

3.0
50
-1

2.5 40 8
40
RI37

2.0 30 6
30
1.5 20 4
20
1.0
10 2
10 0.5
0 0
0 0.0 0 5 10 15 20 25 30
0 5 10 15 20 25 30 35 40 45 50
Time (days)
Time (days)
Fig. 4. Average temperature profile (T) and respiration indices
Fig. 3. Respiration index (RI) and temperature profile (T) in the obtained at 37 °C (RI37) and at the in situ temperature (RIT) during
composting of PCS at pilot plant scale. RI values were calculated from the composting of sludge (DS). RI values were calculated from three
three replicates. replicates.
1158 R.B. Gómez et al. / Bioresource Technology 96 (2005) 1153–1159

70 14 versely, when composting temperatures fell to around

T 20 °C, RIT were lower than RI37. This would indicate
60 RI37 12
that RIT were sensitive enough as to detect the decrease

RI (mg O2*gOM-1*h-1)
RI T
Temperature (ºC)

50 10 in the metabolic activity within the reactor whereas RI37

were less sensitive to these changes. As expected, when
40 8
process temperatures were closer to 37 °C, both indices
30 6 were similar.
In the case of ADS, Fig. 5 shows that RIT and RI37
20 4
followed the same trend as for DS and that RI37 values
10 2 were quite similar for both materials. However when
comparing the two indices, it can be seen that RIT values
0 0
0 5 10 15 20 25 30 were considerably lower for the ADS. This lower activ-
Time (days) ity may be related to the differences in the organic mat-
ter contents of the sludges, since DS contained 80.2%
Fig. 5. Average temperature profile (T) and respiration indices
obtained at 37 °C (RI37) and at the in situ temperature (RIT) during (dry basis) organic matter while ADS contained 61.7%
the composting of anaerobically digested sludge (ADS). RI values were (dry basis). Besides, ADS not only contained less organic
calculated from three replicates. matter but since they were an already digested material,
their organic fraction was less biodegradable than that
of DS. Therefore, activity is not only related to the
metabolism results in the generation of heat and a new organic matter content of the material but to its nature
temperature increase. However, it has also been argued as well.
that this behaviour maybe because high temperatures The differences in RIT values for DS and ADS are in
may lead to a sudden decrease in the metabolic activity good agreement with the differences in the composting
of the microorganisms and consequently, a reduction in profiles of the two sludges. Higher activity and sharper
the amount of heat generated. The material then cools increase and higher temperatures are observed at the
down and temperature drops. This cooling down reacti- beginning of the process for DS (Fig. 4) when compared
vates the metabolic activity, heat is generated again and with the composting of ADS (Fig. 5). A faster drop in
so temperature rises again. This last argument could activity is also observed. The main reason for this differ-
explain the behaviour of the DS since the drop in tem- ence seemed to be the different organic contents and bio-
perature was quicker than for ADS and its organic mat- degradabilities of the two materials.
ter content was higher (80.2% dry basis) than for ADS Results then indicate that RIT are more sensitive to
(61.7% dry basis). However, in practice it is very difficult temperature changes and composition variations of the
to determine the actual reason for the temperature composting material than RI37. Therefore, RIT would
profiles. provide more representative information of the actual
Figs. 4 and 5 also show the respiration indexes ob- metabolic activity in the composter. However, when
tained at 37 °C (RI37) and at the in situ temperature process temperature falls below optimum mesophilic
(RIT) along the process for sludge (DS) and anaerobi- conditions, although RIT are able to detect the actual
cally digested sludge (ADS), respectively. From these metabolic activity in the reactor, they are unable to indi-
Figures it can be seen that both indices correlated well cate the potential metabolic activity of the material since
with temperature evolution. The values indicate a higher their values would correspond to a stable material,
metabolic activity at the beginning of the process, a which is not the case in practice. Therefore under these
reduction in activity during the cooling down phase conditions RIT are useful for the monitoring of the com-
and an increase in activity once the temperature started posting process but not for determining the actual sta-
to increase again. Also, indices were fairly constant bility of the material. Moreover, from the operational
when temperature changes were small. point of view RIT assays are faster than RI37 since incu-
Nevertheless, although RI37 and RIT values followed bation time is only 4 h compared to the 18 required for
the same trend, several differences are found between the latter.
them. In the case of DS (Fig. 4), when temperatures in While respirometry techniques have been widely used
the composter were high, RIT were significantly higher as stability indicators for compost, their use for the
than RI37 and as temperature dropped quickly so did monitoring of the process has been limited. Results pre-
RIT. The reason for this could be that at high tempera- sented here demonstrate that they are actually an excel-
tures prevailing population in the composter was ther- lent indicator for the monitoring of the process although
mophilic. Therefore, this population would show they are far more complicated than temperature mea-
higher activity for assays performed at the actual tem- surement and monitoring. Nevertheless, relationships
peratures of the reactor compared to the activity shown between temperature and respiration indexes are cur-
at 37 °C despite the incubation prior to the assay. Con- rently under investigation.
 R.B. Gómez et al. / Bioresource Technology 96 (2005) 1153–1159 1159

4. Conclusions Garcı́a, C., Polo, A., 1999. Estudio de parámetros bioquı́micos en

procesos de estabilización de residuos orgánicos urbanos. Residuos
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A reliable device has been constructed for determin- Garcı́a, C., Hernández, T., Costa, F., Ayuso, M., 1992. Evaluation of
ing the RI of compost samples. Results have shown that the maturity of municipal waste compost using simple chemical
RI measurements at the in situ temperature of the pro- parameters. Commun. Soil Sci. Plant Anal. 23, 1501–1512.
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the composter. Measurements at 37 °C could lead to Application of experimental design techniques to the optimization
of bench-scale composting conditions of municipal raw sludge.
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