Waste Management 34 (2014) 1546–1552

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Waste Management
journal homepage: www.elsevier.com/locate/wasman

Emissions of ammonia and greenhouse gases during combined

pre-composting and vermicomposting of duck manure
Jinzhi Wang a,b, Zhengyi Hu b,⇑, Xingkai Xu c, Xia Jiang a, Binghui Zheng a, Xiaoning Liu b, Xubin Pan d,
Paul Kardol e
a
State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environment Sciences, Beijing 100012, China
b
College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China
c
State Key Laboratory of Atmospheric Boundary Layer Physics and Atmospheric Chemistry, Institute of Atmospheric Physics, Chinese Academy of Sciences, Beijing 100029, China
d
Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China
e
Department of Forest Ecology and Management, Swedish University of Agricultural Sciences, S 90183 Umeå, Sweden

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

Article history: Combined pre-composting and vermicomposting has shown potential for reclamation of solid wastes,
Received 10 September 2013 which is a significant source of ammonia (NH3), and greenhouse gases (GHG), including nitrous oxide
Accepted 10 April 2014 (N2O), methane (CH4), and carbon dioxide (CO2). Earthworms and amendments may both affect phys-
Available online 15 May 2014
ico-chemical characteristics that control gas-producing processes, and thus affect NH3 and GHG emis-
sions. Here, we used two-way ANOVA to test the effects of addition of reed straw and combined
Keywords: addition of reed straw and zeolite on NH3 and GHG emissions during pre-composting of duck manure,
Ammonia
either with or without a follow-up phase of vermicomposting. Results showed that cumulative N2O,
Greenhouse gas
Reed Straw
CH4, and CO2 emissions during pre-composting and vermicomposting ranged from 92.8, 5.8, and
Zeolite 260.6 mg kg1 DM to 274.2, 30.4, and 314.0 mg kg1 DM, respectively. Earthworms and amendments sig-
Earthworm nificantly decreased N2O and CH4 emissions. Emission of CO2 was not affected by earthworms, but
Composting increased in responses to addition of reed straw. Cumulative NH3 emission ranged from 3.0 to 8.1 g kg1
DM, and was significantly decreased by reed straw and zeolite addition. In conclusion, combined pre-
composting and vermicomposting with reed straw and zeolite addition would be strongly recommended
in mitigating emissions of N2O, CH4, and NH3 from duck manure. Moreover, this method also provides
nutrient-rich products that can be used as a fertilizer.
Ó 2014 Elsevier Ltd. All rights reserved.

1. Introduction Recently, interest in the use of vermicomposting, i.e., using

The reed wetland of Baiyangdian catchment in China is an
important duck farm base with about 8 million ducks, which
produce more than 213,300 tons manure each year. Most of the
duck manure ends up in the water system, causing eutrophication
of surface water. Traditional composting of organic wastes com-
prises a short period of high temperature followed by a period of
low temperature, as such facilitating pathogen reduction, waste
stabilization, as well as mass reduction (Barrington et al., 2002;
Nair et al., 2006). However, the major problems associated with
the traditional composting of organic wastes are the long duration
of the process, losses of nutrients during the prolonged composting
process, frequent aeration required, and a heterogeneous end-
product (Nair et al., 2006; Ndegwa and Thompson, 2001).
⇑ Corresponding author. Tel.: +86 10 88256542; fax: +86 10 88256415.
E-mail address: zhyhu@ucas.ac.cn (Z. Hu).
earthworms to breakdown organic materials, has increased
(Ndegwa and Thompson, 2000; Pramanik et al., 2007). In its basic
form, vermicomposting is a low-cost technology which offers a
higher quality product (known as ‘‘vermicomposts’’) than tradi-
tional composting. Earthworms accelerate composting by biotur-
bation and aeration, and by that, produce an end-product that is
more nutritional (Albanell et al., 1988; Atiyeh et al., 2000), more
fragmented, and has higher microbial activity (Héry et al., 2008;
Vivas et al., 2009). However, a main disadvantage of vermicompo-
sting is that temperature in the feed substrate must be maintained
below 35 °C to avoid the death of earthworms. The temperature
during vermicomposting process is therefore not high enough for
effective pathogen control (EPA, 2003). A combined system that
integrate pertinent attributes from both traditional composting
and vermicomposting has shown potential for reclamation of solid
wastes, not only shortening the stabilization time (Frederickson
et al., 1997), but also improving the quality of end-products as

http://dx.doi.org/10.1016/j.wasman.2014.04.010
0956-053X/Ó 2014 Elsevier Ltd. All rights reserved.
 J. Wang et al. / Waste Management 34 (2014) 1546–1552
fertilizers (Ndegwa and Thompson, 2001). Moreover, vermicompo-
sting also produces earthworms as a fine fodder for poultry with
high protein.
The combined composting and vermicomposting process in the
treatment of poultry manure inevitably involves emissions of
ammonia (NH3) and greenhouse gases (GHG), including nitrous
oxide (N2O), carbon dioxide (CO2), and methane (CH4), which can
contribute to global warming and stratospheric ozone depletion
(Crutzen, 1970). Deposition of emitted NH3 in aquatic and terres-
trial systems can cause eutrophication and soil acidification,
respectively (Metcalfe et al., 1998). Although earthworms hardly
produce any NH3 and GHG themselves, they can significantly influ-
ence physico-chemical properties of the feeding substrate (Hobson
et al., 2005), as such indirectly affecting gas-producing processes,
and thereby affecting NH3 and GHG emissions. Effects of earth-
worms on gas emissions are complicated and no consensus has
been reached. For example, Lubbers et al. (2013) showed that
earthworms increased emissions of N2O and CO2 from soils by
42% and 33%, respectively. Hobson et al. (2005) showed a signifi-
cant effect of earthworms on the efflux of N2O from household
waste, but only a marginal effect on CH4 emission. Others showed
that earthworms significantly increased CO2 emission, but did not
affect N2O fluxes from soils (Chapuis-Lardy et al., 2010; Speratti
and Whalen, 2008). Similarly, Chan et al. (2011) showed that ver-
micomposting of household waste emitted more CO2 and CH4, but
less N2O than traditional composting.
Amendments can be used to change the carbon and nitrogen
content, aerobic conditions, pH, and the adsorbent capacity of
manure and hence mitigate NH3 and GHG emissions. Carbon-rich
organic materials have been used as amendments to increase aer-
ation and immobilization of nitrogen in manure, thereby reducing
gas emissions. For example, Kirchmann and Witter (1989) showed
a reduction of NH3 emission from chicken manure amended with
straw, and Yamulki (2006) reported a reduced N2O and CH4 emis-
sions from fresh cattle farmyard manure amended with straw. Zeo-
lite, a porous mineral with a high cation exchange capacity and
affinity for ammonium (NH+4), has used to reduce NH3 emission
from poultry manure (Witter and Kirchmann, 1989). Similarly,
Wang et al. (2012) showed that reed straw applied singly or in
combination with zeolite reduced NH3 and GHG emissions for
composting duck manure.
Most of previous studies on vermicomposting focus on the fea-
sibility of vermicomposting of different organic wastes (Sangwan
et al., 2008), the factors affecting the growth and reproduction rate
of earthworms (Ndegwa et al., 2000), as well as the quality of ver-
micomposts (Pramanik et al., 2007). In addition, several recent
studies have focused on the effects of earthworms on GHG emis-
sions from soils (e.g., Bradley et al., 2012; Lubbers et al., 2013).
However, little is known about the effects of earthworms on gas
emission during vermicomposting of organic wastes, especially
for the poultry manure. The objectives of this experiment were
therefore to assess the effects of earthworms and amendments
on NH3 and GHG (i.e., N2O, CH4, and CO2) emissions during com-
bined pre-composting and vermicomposting of duck manure and
to develop environment friendly and economic method for reason-
able duck manure management.
2. Materials and methods
2.1. Experimental design
Our experiment was carried out at the University of Chinese
Academy of Sciences, Beijing in autumn 2011. Recently deposited
duck manure was collected from a duck farm in Baiyangdian, Hebei
province, China. Our experiment consisted of a traditional
1547
composting phase (hereafter referred to as ‘pre-composting’) and
a subsequent vermicomposting phase. Reed straw and zeolite were
used as amendments. Dried reed straw was provided by a local
farmer and shredded (<1 cm pieces), the organic carbon, total
nitrogen, total phosphorus of reed straw were 451.6, 5.8, and
0.8 g kg1, respectively. The zeolite with 8.4 of pH was bought in
Hebei province from the Luanping zeolite company, and was
ground and sieved (0.178 mm mesh) for use.
During the pre-composting phase, the experiment included
three treatments: (i) duck manure without any amendments (C),
(ii) duck manure with 40% (w/w) reed straw (S), and (iii) 40% (w/
w) reed straw plus 12% (w/w) zeolite (SZ). All amendments were
applied on a dry weight basis. The treatments were replicated
three times, giving a total of 9 experimental units. Mixtures of
10 kg duck manure (38% moisture content) and corresponding
amendments were completely mixed and placed into each of 9
incubation containers (length  width  height, 53  38  32 cm).
After 45 days of pre-composting, from each container, part of
the pre-composting material was transferred to worm-bins
(length  width  height, 31  24  22 cm) for subsequent vermi-
composting. This provided 0.07 m2 of exposed top surface. The ver-
micomposting stage lasted 32 days. Each worm-bin received 120 g
of earthworms (Eisenia fetida, provided by a farmer in Beijing,
China), corresponding with an optimal stocking density of
1.60 kg worms m2 (Ndegwa et al., 2000). Considering the optimal
C/N ratio of 25 for vermicomposting (Ndegwa and Thompson,
2000), more reed straw was added to optimize environmental con-
ditions for the earthworms. Control bins did not receive earth-
worms, but were amended with the same amount of reed straw.
The vermicomposting phase involved the following treatments:
pre-composting (C, S, SZ)  earthworms (absent, present)  3 rep-
licates = 18 experimental units. Over the course of the experiment
no additional food was added at any stage. The moisture level of
the material was maintained at about 60–65% of wet mass
throughout the vermicomposting stage by spraying the surface
with water at two-day intervals. The worm-bins were incubated
in a humid and dark room at approximately 20 °C.
2.2. Measurements of NH3 and GHG emissions during pre-composting
Emissions of NH3, N2O, CO2, and CH4 during pre-composting
were measured by a closed chamber technique (Czepiel et al.,
1996; Wang et al., 2012). Plastic vessels were transformed into
closed chambers by a tight fit lid with two ports for headspace
gas sampling and for measurement of air temperature (Fig. 1a).
The plastic vessels, which had no bottom, inserted directly into
the duck manure mixture about 2 cm below the surface. The aver-
age headspace volume of vessels was maintained at 2.6 l.
Gases measurements were done at 1, 3, 6, 8, 11, 15, 19, 23, 29,
36, and 45 days after the start of the experiment. During each mea-
surement, 30-ml gas samples were taken from each experimental
container after 0, 10, and 20 min using a 50-ml syringe, and then
immediately sent back to the laboratory for analysis of GHG using
a gas chromatograph coupled with a flame ionization detector and
an electron captor detector (Agilent 7890A, USA). Concentration of
NH3 was measured using M4+ gas monitoring instrument (Changz-
hou Nuoji Instrument Co., Ltd., China) during the early stage of pre-
composting when NH3 emission was high. Gas fluxes (F, mg kg1
DM d1, DM = dry matter) were calculated according to Czepiel
et al. (1996):
V dC
F¼q  ð1Þ
M dt
where q is the density of the gas at sample temperature (°C), V is
the air volume of headspace in the container (m3), M is the initial
1548 J. Wang et al. / Waste Management 34 (2014) 1546–1552
Syringe
a Thermometer
Closed lid
Closed chamber
Area captured
Inserted portion
32 cm
Composting container
Duck manure
cm
38
53 cm
Syringe
b Thermometer
Closed Lid
Area captured
22 cm
Worm bins
cm
Duck manure
24
31 cm
Fig. 1. Experimental set-up for pre-composting (a) and vermicomposting (b) of
duck manure (not according to scale).
dry mass of materials (kg), and dC dt
is the rate of change in gas
concentration in relation to time after the headspace was closed.
To calculate the cumulative NH3, N2O, CO2, and CH4 emissions,
we summed daily values to get the total cumulative gases emis-
sions during the whole experimental period (Wang et al., 2012).
To assess the effects of different treatments on total GHG emis-
sions, measured emissions of N2O, CO2, and CH4 were converted
into CO2-equivalents using global warming potentials of 298 and
25 for N2O and CH4, respectively (IPCC, 2007), and summed to give
total GHG emissions.
2.3. Measurement of GHGs emissions during vermicomposting
Gas sampling and analyses were done similarly as during the
pre-composting stage, except that the worm-bins for vermicompo-
sting had their own tight fit lid with two ports for headspace gas
sampling and enclosed air temperature measurements (Fig. 1b).
Headspace gases were sampled at 18, 25, and 32 days after the
start of vermicomposting. Up to 95% NH3 was emitted during the
first 10 days of pre-composting; hence, we did not measure NH3
emissions during the following vermicomposting.
2.4. Measurements of physical and chemical properties
During the pre-composting and the following vermicomposting
stages, samples of the manure mixtures were collected for mea-
surements of physical and chemical properties. Samples were col-
lected at 0, 11, 23, 36, 45 days, and by the end of experiment for
determination of total organic carbon, total nitrogen, pH, dry mat-
ter content, NH+4–N, and NO 3 –N using national standard methods
(Lu, 1999). Before samples were collected, manure mixtures were
turned and homogenized. Air temperature and temperature in
the center of the duck manure were measured continuously with
HOBO U12-006 temperature logger (Onset Computer Corporation,
USA).
2.5. Statistical analyses
Effects of earthworms and amendments on the physico-chemi-
cal characteristics of duck manure mixtures and on gas emissions
were analyzed by two-way ANOVA. Statistical tests were consid-
ered significant at P < 0.05 level. When effects were significant,
LSD post-hoc tests were used to check for differences among indi-
vidual treatments. For all ANOVA’s, the assumption of normality
was checked with Kolmogorov–Smirnov tests and the assumptions
of homogeneity of variances was checked using Levene’s test. If the
assumptions were not met, data were log-transformed prior to
analysis. Statistical analyses were performed using SAS procedure
GLM-LSD (least significant difference) (SAS version 8.1, SAS Insti-
tute Inc., Cary, NC, USA).
3. Results and discussion
3.1. Ammonia
Across treatments, 81% to 95% of NH3 was emitted during the
first 10 days after the start of the experiment, the emission rates
decreased sharply thereafter (Fig. 2a). Compared with the control,
addition of reed straw (S) and reed straw combined with zeolite
(SZ) significantly reduced cumulative NH3 emission by 48% and
63%, respectively (Table 1).
As an easily decomposable and carbon-rich material, reed straw
increases the amount of degradable carbon which promotes immo-
bilization of ammonium into microbial biomass (Sommer et al.,
2006), and increases the C/N ratio and aeration in manure
(Yamulki, 2006). Therefore, addition of reed straw may increase
the potential for reducing the NH3 emission from duck manure
in comparison to the control (Wang et al., 2012; Yamulki, 2006).
Compared with the control, addition of reed straw combined with
zeolite have more potential for reducing NH3 emission, which was
consistent with the previous findings because of the high affinity of
zeolite for NH+4 (Wang et al., 2012).
3.2. Nitrous oxide
During pre-composting, rates of N2O emission were high during
the first 15 days after the start of the experiment, and decreased
thereafter (Fig. 2b). During following vermicomposting, N2O flux
was significantly affected by the presence of earthworms, sampling
date, and amendments (Fig. 3a). After the whole experiment,
cumulative N2O emission was decreased by earthworms
(P = 0.038) and addition of amendments (P < 0.0001), but there
was no interaction effect between earthworms and amendments
(Table 2). Cumulative N2O emissions for C, S, and SZ treatments
in presence of earthworms respectively decreased by 7%, 43%,
and 28% compared to C, S, and SZ treatments in absence of earth-
worms (Table 1).
The results in this study were consistent with some previous
studies that found a decreased N2O emission in presence earth-
worms (Chan et al., 2011; Contreras-Ramos et al., 2009). However,
Matthies et al. (1999) have shown positive effects of earthworms
on N2O emission from soils. Luth et al. (2011) suggest that there
is probably a threshold of input of available organic nitrogen that
will determine whether earthworms will increase or decrease
N2O emissions. They studied the effect of earthworms on N2O
emission with various amount of pig slurry and showed that earth-
worms increased the N2O emission when supplied pig slurry of
2 L day1, while earthworms decreased N2O emission when sup-
plied 6 and 28 L day1. Increased N2O emission by earthworms in
lower nitrogen environment can be explained by gut content richer
in available nitrogen and carbon compared to outside substrate
 J. Wang et al. / Waste Management 34 (2014) 1546–1552 1549

10 350
C a c

Cumulative NH3 emission (g kg-1)

Cumulative CO2 emission (g kg-1)

S 300
8 SZ
250
6 200

150
4
100
2
50

0 0

0 10 20 30 40 0 10 20 30 40

100 8
b d
Cumulative N2O emission (mg kg-1)

Cumulative CH4 emission (mg kg-1)

X Data
80
6
60

4
40

20 2

0
0
-20
0 10 20 30 40 0 10 20 30 40
Days Days

Fig. 2. Cumulative NH3, N2O, CO2, and CH4 emissions during pre-composting of duck manure alone (C), with addition of reed straw (S), and with addition of reed straw and
zeolite (SZ). Data are mean ± SE.

Table 1
Cumulative NH3 and greenhouse gas emissions during pre-composting and vermicomposting. Data are mean ± SE. C = control, S = addition of reed straw, SZ = addition of reed
straw and zeolite; VC, VS, and VSZ represent the corresponding treatments in presence of earthworms.

Process Treatments NH3 CO2 N2O CH4 N2O + CH4a N2O + CH4 + CO2a
(g kg1 DM) (g kg1 DM) (g kg1 DM) (g kg1 DM) (g kg1 CO2-eq. DM) (g kg1 CO2-eq. DM)
Pre-composting C 8.1 ± 0.6a 237.8 ± 19.0b 80.6 ± 4.1a 5.7 ± 1.5a 24.2 ± 1.3a 262.0 ± 20.2a
S 4.2 ± 0.1b 280.8 ± 8.8ab 37.8 ± 0.9b 3.5 ± 0.4ab 11.4 ± 0.3c 292.2 ± 8.8a
SZ 3.0 ± 0.0b 288.6 ± 6.6a 59.5 ± 7.6a 1.9 ± 0.8b 17.8 ± 2.2b 306.4 ± 6.8a
VC 8.1 ± 0.6a 237.8 ± 19.0b 80.6 ± 4.1a 5.7 ± 1.5a 24.2 ± 1.3a 262.0 ± 20.2a
VS 4.2 ± 0.1b 280.8 ± 8.8ab 37.8 ± 0.9b 3.5 ± 0.4ab 11.4 ± 0.3c 292.2 ± 8.8a
VSZ 3.0 ± 0.0b 288.6 ± 6.6a 59.5 ± 7.6a 1.9 ± 0.8b 17.8 ± 2.2b 306.4 ± 6.8a
Vermicomposting C 22.8 ± 1.4 193.6 ± 21.6 24.7 ± 7.8 58.3 ± 6.4 81.1 ± 6.4
S 30.2 ± 0.3 96.0 ± 6.0 15.7 ± 7.7 29.0 ± 1.6 59.2 ± 1.8
SZ 25.4 ± 0.4 69.6 ± 9.4 7.9 ± 3.1 20.9 ± 2.8 46.3 ± 2.6
VC 26.6 ± 0.3 180.1 ± 9.9 4.8 ± 1.3 53.8 ± 3.0 80.4 ± 2.6
VS 27.8 ± 0.5 55.0 ± 13.3 2.3 ± 0.8 16.4 ± 4.0 44.3 ± 4.1
VSZ 23.9 ± 0.1 50.4 ± 10.8 4.0 ± 2.2 15.1 ± 3.2 39.0 ± 3.2
Pre-composting + vermicomposting C 260.6 ± 19.6 274.2 ± 25.0 30.4 ± 6.3 82.5 ± 7.4 343.0 ± 26.4
S 311.0 ± 9.0 133.8 ± 6.2 19.2 ± 7.4 40.4 ± 1.7 351.4 ± 10.6
SZ 314.0 ± 6.4 129.0 ± 11.0 9.8 ± 2.7 38.7 ± 3.3 352.7 ± 9.2
VC 264.4 ± 19.2 260.7 ± 6.0 10.5 ± 1.8 77.9 ± 1.7 342.3 ± 17.7
VS 308.7 ± 8.8 92.8 ± 12.9 5.8 ± 0.6 27.8 ± 3.8 336.5 ± 5.5
VSZ 312.5 ± 6.5 109.9 ± 3.4 5.9 ± 3.0 32.9 ± 1.1 345.4 ± 7.1

Values in the same column followed by the same letters indicate no significant differences at P < 0.05 level.
a
Cumulative greenhouse gas emissions were expressed in CO2-equivalents, using global warming potential of 1, 25 and 298 for CO2, CH4 and N2O, respectively (IPCC,
2007).

(e.g., soils) in which earthworms lived and fed (Horn et al., 2003). with higher nitrogen was reduced by earthworms during pre-com-
This richer environment induces a microbial activity of denitrifica- posting and vermicomposting, a possible explanation was that N2O
tion higher inside the gut than outside, and thus enhances N2O emission from earthworm gut was probably offset by the reduction
emissions (Frederickson and Howell, 2003; Hobson et al., 2005). of anaerobic denitrification due to the burrowing action of the
On the other hand, decreased N2O emission by earthworms in earthworms (Chan et al., 2011). Compared with the control, reed
nitrogen-rich substrates can be explained by a gut content poorer straw and its combination with zeolite significantly decreased
in nitrogen content compared to the outside substrate (Luth et al., cumulative N2O emissions during the whole experiment (Table 1),
2011). In this study, cumulative N2O emission from duck manure which was in line with the findings of Wang et al. (2012), who
1550 J. Wang et al. / Waste Management 34 (2014) 1546–1552

18 80
16 a Day 18 Air
Day 25 C
N2O flux (mg kg d )

70
-1

14 S
Day 32
-1

12 SZ
60

Temperature ( o C)
10
8 50
6
4 40
2
0 30
C S SZ VC VS VSZ
20
1600
1400 b 10
0 10 20 30 40
CO2 flux (mg kg d )
-1

1200 Days
-1

1000
Fig. 4. Temporal dynamics of air temperature and temperature in the center of
800 duck manure for the control (C), reed straw (S), reed straw and zeolite (SZ)
treatments. Down arrows indicate duck manure turning. The horizontal dashed line
600
at 55 °C marks the threshold temperature for pathogen kill.
400
amendments (Table 2). Compared with the control, addition of
200
reed straw, whether or not combined with zeolite, increased
0 cumulative CO2 emission by approximate 20%, which might result
C S SZ VC VS VSZ
from that reed straw caused a relatively long duration of higher
1.6 temperature that promoted the decomposition of duck manure
(Fig. 4). Earthworms had no effect on the cumulative CO2 emission
1.4 c
in this study (Tables 1 and 2), which contrasted with other studies
CH4 flux (mg kg d )
-1

1.2 that the presence of earthworms increased CO2 emission from soils
-1

1.0 (Chapuis-Lardy et al., 2010; Lubbers et al., 2013). As a whole, the

.8 contrasted findings might result from the different physico-
chemical properties of substrates (e.g., the status of available
.6
carbon pools) in which earthworms lived and fed. For example,
.4 earthworms may be able to accelerate the decomposition of carbon
.2 in soils with lower available carbon content and thus enhance CO2
emission. In substrates with larger, more available carbon pools,
0.0
C S SZ VC VS VSZ the earthworm effect may be eclipsed by overall higher decompo-
Absence of earthworms Presence of earthworms sition rates (Lubbers et al., 2013).
Treatments
3.4. Methane
Fig. 3. Fluxes of N2O, CO2, and CH4 during vermicomposting of duck manure alone
(C), with addition of reed straw (S), and with addition of reed straw and zeolite (SZ),
in presence or absence of earthworms. Data are mean ± SE. During pre-composting and vermicomposting, cumulative CH4
emission was decreased by earthworms (P = 0.005) (Table 2).
Cumulative CH4 emissions for C, S, and SZ treatments in presence
Table 2 of earthworms respectively decreased by 65%, 70%, and 40% com-
Results from two-way ANOVA analysis for the effects of earthworms (E) and pared to C, S, and SZ treatments in absence of earthworms (Table 1).
amendments (A) as fixed factors on GHG emissions during the whole stages of Earthworms are ecosystem engineers, they affect physico-chemical
composting and subsequent vermicomposting.
characteristics of substrates and interact with microbes through
Treatments CO2 N2O CH4 N2O + CH4 N2O + CH4 + CO2 their feeding, burrowing and casting activity (Atiyeh et al., 2000;
E 0.00ns 5.42⁄ 12.15⁄⁄ 5.99⁄ 0.41ns Hobson et al., 2005; Koubová et al., 2012), and by that affect gases
A 9.71⁄⁄ 91.46⁄⁄⁄ 4.31⁄ 94.06⁄⁄⁄ 0.11ns emissions. For example, in our study earthworms significantly
EA 0.03ns 0.63ns 1.71ns 0.64ns 0.12ns decreased cumulative CH4 emission as the net result of CH4 pro-
⁄, ⁄⁄, ⁄⁄⁄
: Statistically significant at P < 0.05, 0.01, 0.001, respectively. duction and CH4 oxidation. Koubová et al. (2012) showed that
ns: not statistically significant. the abundance of the most common methanogen Methanosaricina
in a cattle-impacted soil decreased by earthworms which
found that reed straw can create more aerobic conditions in the increased aeration by their burrowing activity. On the other hand,
duck manure, and thus increasing the potential for reducing N2O the earthworms may have stimulated methanotrophic bacteria and
emissions. by that increased methane oxidation rates (Héry et al., 2008). The
aforementioned results may explain the decreased CH4 emission in
this study compared to composting without earthworms.
3.3. Carbon dioxide

Emissions of CO2 peaked within the first 15 days after the start 3.5. Total greenhouse gas emissions
of the experiment (Fig. 2c), and more than 90% of CO2 was emitted
during pre-composting (Table 1). Cumulative CO2 emission during Emissions of N2O, CO2, and CH4 measured during pre-compost-
the whole experiment was significantly increased by addition of ing and vermicomposting of duck manure were converted to
 J. Wang et al. / Waste Management 34 (2014) 1546–1552 1551

Table 3
Physical and chemical properties of duck manure without amendments (C), with addition of reed straw (S), reed straw and zeolite (SZ) during pre-composting. Data are
mean ± SE.

Treatments Time (d) pH Org. C (g C kg1 DM) TN (g C kg1 DM) C/N NH+4 (g N kg1 DM) NO
3 (g N kg
1
DM)
C 0 7.7 ± 0.1 246.7 ± 4.8 13.4 ± 0.2 18.4 ± 0.4 6.4 ± 0.2 0.4 ± 0.0
11 8.0 ± 0.0 238.6 ± 7.6 11.6 ± 0.0 20.5 ± 0.7 7.6 ± 0.1 0.4 ± 0.1
23 7.9 ± 0.0 211.3 ± 4.4 15.2 ± 0.2 13.9 ± 0.3 3.0 ± 0.1 0.5 ± 0.0
36 8.2 ± 0.0 189.7 ± 0.6 16.2 ± 0.1 11.7 ± 0.1 2.4 ± 0.1 0.5 ± 0.0
45 8.1 ± 0.0 184.6 ± 2.6 15.1 ± 1.5 12.5 ± 1.4 1.6 ± 0.1 0.6 ± 0.0
S 0 7.7 ± 0.0 305.4 ± 4.6 11.3 ± 0.1 27.2 ± 0.7 5.5 ± 0.1 0.5 ± 0.0
11 7.9 ± 0.0 253.4 ± 12.4 9.8 ± 0.2 25.8 ± 1.2 7.4 ± 0.2 0.7 ± 0.1
23 7.8 ± 0.0 248.4 ± 2.5 13.2 ± 0.1 18.8 ± 0.4 2.7 ± 0.1 0.8 ± 0.1
36 7.9 ± 0.0 234.5 ± 5.8 12.7 ± 0.9 18.8 ± 1.9 2.1 ± 0.0 0.8 ± 0.1
45 7.8 ± 0.0 235.5 ± 4.4 12.9 ± 0.2 18.2 ± 0.3 1.1 ± 0.0 0.8 ± 0.0
SZ 0 7.6 ± 0.0 281.4 ± 4.3 10.4 ± 0.1 27.2 ± 0.7 5.7 ± 0.1 0.4 ± 0.1
11 7.8 ± 0.0 241.9 ± 8.4 10.5 ± 1.2 23.5 ± 2.6 7.2 ± 0.4 0.6 ± 0.1
23 7.8 ± 0.0 206.7 ± 1.9 12.7 ± 0.2 16.3 ± 0.4 3.1 ± 0.1 0.7 ± 0.1
36 7.8 ± 0.0 220.3 ± 0.4 12.5 ± 0.4 17.6 ± 0.6 2.3 ± 0.1 0.7 ± 0.1
45 7.7 ± 0.0 208.3 ± 3.4 12.4 ± 0.1 16.8 ± 0.2 1.1 ± 0.1 0.8 ± 0.1

Table 4
Physical and chemical properties of the end products after pre-composting and subsequent vermicomposting. C = control, S = addition of reed straw, SZ = addition of reed straw
and zeolite; VC, VS, and VSZ represent the corresponding treatments in presence of earthworms. Data are mean ± SE.

Treatments pH Org. C (g kg1) TN (g kg1) C/N TP (g kg1) NH+4 (g kg1) NO

3 (g kg
1
)
C 7.5 ± 0.00 223.6 ± 1.1 9.7 ± 0.2 23.0 ± 0.4 9.2 ± 0.4 0.10 ± 0 0.35 ± 0.0
S 7.6 ± 0.01 233.8 ± 3.0 9.3 ± 0.1 25.2 ± 0.5 8.3 ± 0.2 0.12 ± 0 0.22 ± 0.0
SZ 7.5 ± 0.00 227.2 ± 3.3 9.5 ± 1.2 24.0 ± 0.5 8.0 ± 0.2 0.10 ± 0 0.26 ± 0.0
VC 7.2 ± 0.02 213.4 ± 3.6 10.7 ± 0 20.0 ± 0.3 9.8 ± 0.2 0.14 ± 0 0.52 ± 0.0
VS 7.2 ± 0.02 236.7 ± 1.0 9.6 ± 0.1 24.6 ± 0.4 9.3 ± 0.3 0.12 ± 0 0.43 ± 0.0
VSZ 7.0 ± 0.01 233.1 ± 6.6 9.6 ± 0.1 24.4 ± 1.0 8.9 ± 0.2 0.12 ± 0 0.48 ± 0.0

Table 5
Results from two-way ANOVA analysis for the effects of earthworms (E) and amendments (A) as fixed factors on physical and chemical properties of end products after pre-
composting and subsequent vermicomposting.

Treatments pH Org. C TN C/N TP NH+4 NO

3

⁄⁄⁄ ⁄⁄ ⁄ ⁄⁄ ⁄⁄⁄
E 1550.63 0.02ns 16.42 5.19 14.35 24.07 500.64⁄⁄⁄
A 64.42⁄⁄⁄ 11.42⁄⁄ 19.17⁄⁄⁄ 21.40⁄⁄⁄ 7.56⁄⁄ 3.47ns 51.61⁄⁄⁄
EA 1.23ns 2.84ns 4.80⁄ 4.81⁄ 0.17ns 5.07⁄ 2.08ns
⁄, ⁄⁄, ⁄⁄⁄
: Statistically significant at P < 0.05, 0.01, 0.001, respectively.
ns: not statistically significant.

CO2-equivalents, and summed to get total GHG emissions which
ranged from 336.5 to 352.7 g CO2-eq. kg1 DM (Table 1). Earth-
worms did not affect total GHG emissions (Table 2), which was
contrary to the findings that earthworms increased GHG emissions
from soils (Lubbers et al., 2013). In this study, the negligible effects
of earthworms on total GHG emissions mainly resulted from their
effects on CO2 emission, which accounted for up to 92% of the total
GHG emissions. On the other hand, total GHG emissions during the
whole experiment were not affected by amendments, a possible
reason was that increased CO2 emissions caused by amendments
were offset by the reduction of N2O and CH4 emissions.
When CO2 emission as an indicator of biological activity was
excluded from the accounting, total combined N2O–CH4 emission
ranging from 27.8 to 82.5 g CO2-eq. kg1 DM on average, was sig-
nificantly reduced by both earthworms (P = 0.031) and amend-
ments (P < 0.0001) (Table 2). Total combined N2O–CH4 emissions
for C, S, and SZ treatments in presence of earthworms were respec-
tively 6%, 31%, and 15% lower compared to C, S, and SZ treatments
in absence of earthworms. Total combined N2O–CH4 emission was
reduced by reed straw applied singly or in combination with zeo-
lite (Tables 1 and 2). Hence, in terms of GHG emissions, combined
pre-composting and vermicomposting with reed straw and zeolite
addition would be strongly recommended as effective ways to
dispose duck manure.
3.6. Physical and chemical characteristics of duck manure
During pre-composting, the temperature in the duck manure
exceeded the critical temperature for pathogen kill (55 °C) for
two and four consecutive days (EPA, 2003) shortly after the start
of our experiment (Fig. 4). In all treatments, the temperature
decreased over time. After turning the duck manure (see Section
2), the temperature increased immediately; subsequent decrease
in temperature was faster in control than in reed straw treatments.
The pH increased during the first 11 days of pre-composting, and
decreased thereafter (Table 3). Total organic carbon decreased over
time; most organic carbon was lost during the first 23 days of the
experiment (Table 3). Total nitrogen increased over time for all
treatments. Hence, the C/N ratio of the duck manure decreased
over time. For all treatments, the NH+4 concentration in the duck
manure increased during the first 11 days of pre-composting
(Table 3), when the rate of NH+4 production due to intense organic
matter degradation exceeded the rate of NH+4 loss through nitrifica-
tion of NH+4 to NO3 and gaseous NH3 emission (Hao et al., 2004).
1552 J. Wang et al. / Waste Management 34 (2014) 1546–1552
The amount of NH+4 decreased sharply afterward due to the nitrifi-
cation of NH+4 to NO 
3 , therefore gradually increased NO3 concen-
tration in this study.
Table 4 shows the physical and chemical properties of the end
products after the vermicomposting process. Earthworms signifi-
cantly decreased pH (Tables 4 and 5), which may be due to the
accumulation of organic acids from microbial metabolism or from
the production of fulvic and humic acids during decomposition
(Albanell et al., 1988). Concentrations of total nitrogen, total phos-
phorus, NO-3, and NH+4 in the manure mixtures were significantly
increased by earthworms, especially for the NO 3 concentration
(Tables 4 and 5). Similar results were reported by Atiyeh
et al.(2000) who found that earthworms produced conditions that
favored nitrification, therefore increased the NO 3 content of
manure.
4. Conclusions
Earthworms have a positive effect to mitigate total combined
N2O–CH4 emissions as CO2-equivalents and marginal effect on
CO2 emissions during combined pre-composting and vermicompo-
sting of duck manure. Cumulative emissions of NH3, N2O, and CH4
were significantly reduced by the addition of reed straw and zeo-
lite. Therefore, combined pre-composting and vermicomposting
with reed straw and zeolite addition would be a recommended
method to dispose duck manure and reduce NH3 and GHG emis-
sions, as well as to provide nutrient rich products as fertilizers.
Acknowledgement
This work was supported by the Key Projects in the National
Science & Technology Pillar Program (2007BAD87B04-02 and
2012BAJ24B06).
References
Albanell, E., Plaixats, J., Cabrero, T., 1988. Chemical changes during
vermicomposting (Eisenia fetida) of sheep manure mixed with cotton
industrial wastes. Biol. Fertil. Soils 6 (3), 266–269.
Atiyeh, R.M., Domínguez, J., Subler, S., Edwards, C.A., 2000. Changes in biochemical
properties of cow manure during processing by earthworms (Eisenia andrei,
Bouché) and the effects on seedling growth. Pedobiologia 44 (6), 709–724.
Barrington, S., Choinière, D., Trigui, M., Knight, W., 2002. Effect of carbon source on
compost nitrogen and carbon losses. Bioresour. Technol. 83 (3), 189–194.
Bradley, R.L., Chroňáková, A., Elhottová, D., Šimek, M., 2012. Interactions between
land-use history and earthworms control gross rates of soil methane production
in an overwintering pasture. Soil Biol. Biochem. 53, 64–71.
Chan, Y.C., Sinha, R.K., Wang, W., 2011. Emission of greenhouse gases from home
aerobic composting, anaerobic digestion and vermicomposting of household
wastes in Brisbane (Australia). Waste Manage. Res. 29 (5), 540–548.
Chapuis-Lardy, L., Brauman, A., Bernard, L., Pablo, A.L., Toucet, J., Mano, M.J., Weber,
L., Brunet, D., Razafimbelo, T., Chotte, J.L., Blanchart, E., 2010. Effect of the
endogeic earthworm Pontoscolex corethrurus on the microbial structure and
activity related to CO2 and N2O fluxes from a tropical soil (Madagascar). Appl.
Soil Ecol. 45 (3), 201–208.
Contreras-Ramos, S.M., Álvarez-Bernal, D., Montes-Molina, J.A., Van Cleemput, O.,
Dendooven, L., 2009. Emission of nitrous oxide from hydrocarbon contaminated
soil amended with waste water sludge and earthworms. Appl. Soil Ecol. 41 (1),
69–76.
Crutzen, P.J., 1970. The influence of nitrogen oxides on the atmospheric ozone
content. Q. J. R. Meteorol. Soc. 96 (408), 320–325.
Czepiel, P., Douglas, E., Harriss, R., Crill, P., 1996. Measurements of N2O from
composted organic wastes. Environ. Sci. Technol. 30 (8), 2519–2525.
Epa, U.S., 2003. Environmental Regulations and Technology: Control of Pathogens
and Vector Attraction in Sewage Sludge. United States Environment Protection
Agency, Cincinnati, OH.
Frederickson, J., Butt, K.R., Morris, R.M., Daniel, C., 1997. Combining vermiculture
with traditional green waste composting systems. Soil Biol. Biochem. 29 (3–4),
725–730.
Frederickson, J., Howell, G., 2003. Large-scale vermicomposting: emission of nitrous
oxide and effects of temperature on earthworm populations. Pedobiologia 47
(5–6), 724–730.
Héry, M., Singer, A.C., Kumaresan, D., Bodrossy, L., Stralis-Pavese, N., Prosser, J.I.,
Thompson, I.P., Murrell, J.C., 2008. Effect of earthworms on the community
structure of active methanotrophic bacteria in a landfill cover soil. ISME J. 2 (1),
92–104.
Hao, X., Chang, C., Larney, F.J., 2004. Carbon, nitrogen balances and greenhouse gas
emission during cattle feedlot manure composting. J. Environ. Qual. 33 (1), 37–
44.
Hobson, A., Frederickson, J., Dise, N., 2005. CH4 and N2O from mechanically turned
windrow and vermicomposting systems following in-vessel pre-treatment.
Waste Manage. (Oxford) 25 (4), 345–352.
Horn, M.A., Schramm, A., Drake, H.L., 2003. The earthworm gut: an ideal habitat for
ingested N2O-producing microorganisms. Appl. Environ. Microbiol. 69 (3),
1662–1669.
IPCC, 2007. Climate change 2007: mitigation of climate change. In: Contribution of
Working Group III to the Fourth Assessment Report of the Intergovernmental
Panel on Climate Change. Cambridge University Press, Cambridge, United
Kingdom, New York, NY, USA.
Kirchmann, H., Witter, E., 1989. Ammonia volatilization during aerobic and
anaerobic manure decomposition. Plant Soil 115 (1), 35–41.
Koubová, A., Goberna, M., Šimek, M., Chroňáková, A., Pižl, V., Insam, H., Elhottová, D.,
2012. Effects of the earthworm Eisenia andrei on methanogens in a cattle-
impacted soil: a microcosm study. Eur. J. Soil Biol. 48, 32–40.
Lu, R., 1999. Soil Agrochemistry Analysis Method. China Agricultural Science and
Technology Press, Beijing (in Chinese).
Lubbers, I.M., van Groenigen, K.J., Fonte, S.J., Six, J., Brussaard, L., van Groenigen, J.W.,
2013. Greenhouse-gas emissions from soils increased by earthworms. Nat. Clim.
Change 3, 187–194.
Luth, Robin, P., Germain, P., Lecomte, M., Landrain, B., Li, Y., Cluzeau, D., 2011.
Earthworm effects on gaseous emissions during vermifiltration of pig fresh
slurry. Bioresour. Technol. 102 (4), 3679–3686.
Matthies, C., Schmittroth, M., Drake, H.L., 1999. Evidence for involvement of gut-
associated denitrifying bacteria in emission of nitrous oxide (N2O) by
earthworms obtained from garden and forest soils. Appl. Environ. Microbiol.
65 (8), 3599–3604.
Metcalfe, S.E., Derwent, R.G., Whyatt, J.D., Dyke, H., 1998. Nitrogen deposition and
strategies for the control of acidification and eutrophication across Great
Britain. Water Air Soil Pollut. 107 (1), 121–145.
Nair, J., Sekiozoic, V., Anda, M., 2006. Effect of pre-composting on vermicomposting
of kitchen waste. Bioresour. Technol. 97 (16), 2091–2095.
Ndegwa, P., Thompson, S., 2001. Integrating composting and vermicomposting in
the treatment and bioconversion of biosolids. Bioresour. Technol. 76 (2), 107–
112.
Ndegwa, P., Thompson, S., Das, K., 2000. Effects of stocking density and feeding rate
on vermicomposting of biosolids. Bioresour. Technol. 71 (1), 5–12.
Ndegwa, P.M., Thompson, S., 2000. Effects of C-to-N ratio on vermicomposting of
biosolids. Bioresour. Technol. 75 (1), 7–12.
Pramanik, P., Ghosh, G., Ghosal, P., Banik, P., 2007. Changes in organic-C, N, P and K
and enzyme activities in vermicompost of biodegradable organic wastes under
liming and microbial inoculants. Bioresour. Technol. 98 (13), 2485–2494.
Sangwan, P., Kaushik, C., Garg, V., 2008. Feasibility of utilization of horse dung
spiked filter cake in vermicomposters using exotic earthworm Eisenia foetida.
Bioresour. Technol. 99 (7), 2442–2448.
Sommer, S.G., Zhang, G.Q., Bannink, A., Chadwick, D., Misselbrook, T., Harrison, R.,
Hutchings, N., Menzi, H., Monteny, G., Ni, J., 2006. Algorithms determining
ammonia emission from buildings housing cattle and pigs and from manure
stores. Adv. Agron. 89, 261–335.
Speratti, A.B., Whalen, J.K., 2008. Carbon dioxide and nitrous oxide fluxes from soil
as influenced by anecic and endogeic earthworms. Appl. Soil Ecol. 38 (1), 27–33.
Vivas, A., Moreno, B., Garcia-Rodriguez, S., Benitez, E., 2009. Assessing the impact of
composting and vermicomposting on bacterial community size and structure,
and microbial functional diversity of an olive-mill waste. Bioresour. Technol.
100 (3), 1319–1326.
Wang, J.Z., Hu, Z.Y., Zhou, X.Q., An, Z.Z., Gao, J.F., Liu, X.N., Jiang, L.L., Lu, J., Kang, X.M.,
Li, M., Hao, Y.B., Kardol, P., 2012. Effects of reed straw, zeolite, and
superphosphate amendments on ammonia and greenhouse gas emissions
from stored duck manure. J. Environ. Qual. 41 (4), 1221–1227.
Witter, E., Kirchmann, H., 1989. Peat, zeolite and basalt as adsorbents of
ammoniacal nitrogen during manure decomposition. Plant Soil 115 (1), 43–52.
Yamulki, S., 2006. Effect of straw addition on nitrous oxide and methane emissions
from stored farmyard manures. Agric. Ecosyst. Environ. 112 (2–3), 140–145.