Soil Biology & Biochemistry 88 (2015) 294e297

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Soil Biology & Biochemistry

journal homepage: www.elsevier.com/locate/soilbio

Short communication

Interactions between earthworms and mesofauna has no significant

effect on emissions of CO2 and N2O from soil
Haitao Wu*, Mingzhu Lu, Xianguo Lu, Qiang Guan, Xinhua He*
Key Laboratory of Wetland Ecology and Environment, Institute of Northeast Geography and Agroecology, Chinese Academy of Sciences, Changchun,
Jilin 130012, China

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

Article history: Soil fauna can significantly affect soil CO2 and N2O emissions, but little is known about interactions
Received 8 January 2015 between faunal groups and their relative contribution to such emissions. Over a 64-day microcosm in-
Received in revised form cubation, we studied the effects of an epigeic earthworm (Eisenia fetida), mesofauna (Collembola plus
1 May 2015
oribatid mites) and their combinations on soil CO2 and N2O emissions under two faunal densities.
Accepted 1 June 2015
Available online 17 June 2015
Earthworms significantly enhanced soil CO2 and N2O emissions, while mesofauna only increased N2O
emissions. Soil CO2 and N2O emissions were significantly affected by earthworm density, but not by
mesofauna density. No significant interactive effects between earthworms and mesofauna were found on
Keywords:
Carbon
soil CO2 and N2O emissions. Our results indicate that earthworms probably play the dominant roles in
Greenhouse gas determining soil CO2 and N2O emissions where they coexist with soil mesofauna.
Interaction © 2015 Elsevier Ltd. All rights reserved.
Nitrogen cycling
Soil fauna

Soils act as important sources or sinks for carbon dioxide (CO2)
and nitrous oxide (N2O), both well-known greenhouse gases.
Globally, ~20% of CO2 and 62% of N2O emissions originate from soils
(Rastogi et al., 2002; IPCC, 2007). Soil fauna can either suppress,
delay, increase or accelerate soil CO2 and N2O emissions depending
on the group through their effects on the processes of decompo-
sition, nitrification and denitrification (Frouz et al., 2007; Kuiper
et al., 2013; Wu et al., 2013). Numerous studies have confirmed
that earthworms affect soil CO2 and N2O emissions (Rizhiya et al.,
2007; Chapuis-Lardy et al., 2010; Giannopoulos et al., 2010; Lub-
bers et al., 2013; Frouz et al., 2014), through their direct and indirect
impacts on the soil environment, the quality of resources and mi-
crobial processes (Drake and Horn, 2006; Speratti and Whalen,
2008; Nebert et al., 2011; Lubbers et al., 2013). A small number of
studies have also shown effects of mesofauna, such as Collembola
and Acarina, on soil CO2 emissions (Fox et al., 2006; Wickings and
Grandy, 2011), but very few studies measured N2O emissions. Lit-
tle information is available on the interactive effects among soil
faunal groups on soil CO2 and N2O emissions (Collison et al., 2013;
Kuiper et al., 2013; Thakur et al., 2014). The objectives of this study
* Corresponding authors. 4888 Shengbei Street, Changchun, Jilin 130012, China.
Tel.: þ86 431 85542272; fax: þ86 431 85542298.
E-mail addresses: wuhaitao@iga.ac.cn (H. Wu), igalmz@126.com (M. Lu), luxg@
iga.ac.cn (X. Lu), jlsd_gq@163.com (Q. Guan), xinhua5658@hotmail.com (X. He).
were to assess the individual effects of earthworms and soil mes-
ofauna, as well as their interactions on soil CO2 and N2O emissions,
in a laboratory-based incubation experiment.
Soil (Gleysol, FAO taxonomy, 3% sand, 48% silt, 49% clay, pH
(CaCl2) 6.0; see Table 3), adult epigeic earthworms (Eisenia fetida,
0.9 ± 0.04 g fresh weight) and mesofauna (oribatid mites plus
Collembola, see their extraction from Frouz et al., 2007) were
collected in June 2010 from their natural community in an un-
inundated scrub-shrub wetland at the Sanjiang Mire Wetland Sta-
tion (47
1300 500 N, 133
1300100 E), Heilongjiang, China. Earthworms
were voided for 48 h to clean their guts before introductions to
microcosms (see Dalby et al., 1996). The Collembola genera
included Ceratophysella, Friesea, Hypogastrura and Isotomurus,
while the oribatid mite genera were Areozetes, Ceratozetes, Dam-
aeus. and Scheloribates.
A microcosm experiment modeled after Thakur et al. (2014) was
conducted between June and August 2010. Sieved (2 mm) soils
were defaunated (heated for 24 h at 65
C) (Kaneda and Kaneko,
2011) and then filled into Kilner jars (500 mL) with 104 g of air-
dried soil and packed to a bulk density of 0.52 g cm3. Soils were
then adjusted to 60% water filled pore space (WFPS). Soil micro-
cosms were pre-incubated at 20
C and 60% humidity for 3 days
(maintained by distilled water every 2 days) to facilitate microbial
colonization in an environment-controlled chamber. Then, earth-
worms, mites and Collembola were added to the soils (Table 1) with

http://dx.doi.org/10.1016/j.soilbio.2015.06.005
0038-0717/© 2015 Elsevier Ltd. All rights reserved.
 H. Wu et al. / Soil Biology & Biochemistry 88 (2015) 294e297 295

Table 1 NHþ 4 , DOC, microbial biomass carbon (MBC) and nitrogen (MBN)
Introduction density of fauna in treatments after soil pre-incubation in microcosms. were as per Wu et al. (2015). The faunal contributions to N2O and
E, earthworm of Eisenia fetida; Meso, mesofauna as oribatid mites (Acari: Oribatid)
plus springtails (Collembola).
CO2 fluxes were taken as the differences between a faunal treat-
ment minus the control. Data (means ± SE, n ¼ 3 or 4) were sub-
Treatment Faunal density jected to ANOVA and significant differences between treatments
Earthworm Mite þ Springtail were compared with the Tukey test at P ¼ 0.05 with the SPSS 21.0
# microcosm1 # m2 # microcosm1 # m2 package. The interactive effects between earthworm and meso-
fauna on CO2 and N2O emissions were analyzed using a two-way
Control e e e e
E1 1 52 e e ANOVA with the presence of earthworms or mesofauna as two
E3 3 156 independent factors.
Meso1 e e 80 þ 60 4160 þ 3120 No dead or newly hatched earthworms were observed and fresh
Meso3 e e 240 þ 180 12480 þ 9360 earthworm biomass decreased by 19.4e22.3% after 64-days incu-
E1 þ Meso1 1 52 80 þ 60 4160 þ 3120
E3 þ Meso3 3 156 240 þ 180 12480 þ 9360
bation. Abundances of oribatid mites and collembolans decreased
by 28.6e30.8% and 11.4e17.5%, respectively. However, no signifi-
cant differences were found for the changing percentages for
earthworm weight losses and mesofauna density decreases be-
seven replicates per treatment (three for gas flux, and four for tween treatments.
faunal extractions and soil analyses). Seven treatments were Earthworms significantly enhanced both soil CO2 (F ¼ 29.5,
included as showed in Table 1. Microcosms were incubated under P < 0.01) and N2O (F ¼ 21.2, P < 0.01) emission rates, while mes-
the same conditions with pre-incubation over 64 days. The septum ofauna only increased N2O emissions (F ¼ 14.8, P < 0.05), compared
seals in Kilner jar lids were kept open between sampling dates, and with the control (Table 2). The contributions to CO2 and N2O
microcosms were covered with woven black cloth to allow gaseous emissions were generally greater for earthworms than for meso-
exchange while preventing faunal escape. No food was added to the fauna (Table 2). The earthworm-induced increases in CO2 fluxes
microcosm system over the incubation (64 days). Extracted CO2 and (38.2e106%, Table 2) were comparable to other studies (Chapuis-
N2O were analyzed using gas chromatography (Agilent 4890, Cal- Lardy et al., 2010; Paul et al., 2012; Crumsey et al., 2013). Howev-
ifornia, USA) accorded to Wu et al. (2015). Fluxes were calculated by er, the 139e386% enhancements to soil N2O emissions were higher
assuming a linear increase of CO2 and N2O within a jar during the than previous studies (Giannopoulos et al., 2010; Lubbers et al.,
closing period. Cumulative CO2 and N2O emissions were estimated 2013), which was probably due to the relatively high N concen-
assuming linear changes between subsequent flux measurements tration (6.1 g kg1 DW) in our soils (Table 3). In agreement with
(Giannopoulos et al., 2010). Analyses of soil total C, total N, NO 3, Kuiper et al. (2013), no significant effects of mesofauna on CO2

Table 2
Effects of fauna on mean CO2 and N2O emission rates over a 64-day microcosm incubation. Fauna effect is the difference between the mean gas emission rates of the fauna
treatment and control. Values (means ± SE, n ¼ 36) followed by the same letter within a column are not statistically different at P < 0.05. Abbreviations see treatment details in
Table 1.

Treatment CO2 (mg C kg soil1 h1) N2O (mg N kg soil1 h1)

CO2 mean flux Fauna-induced increase (%) N2O mean flux Fauna-induced increase (%)

Control 2.92 ± 0.26c e 0.34 ± 0.09e e

E1 4.01 ± 0.38b 38.2 ± 5.91b 0.81 ± 0.16c 139 ± 23.6b
E3 5.98 ± 0.65a 106 ± 12.5a 1.66 ± 0.31b 386 ± 53.5a
Meso1 3.22 ± 0.30c 9.67 ± 3.46c 0.51 ± 0.11d 60.6 ± 8.99c
Meso3 3.25 ± 0.28c 10.2 ± 2.01c 0.59 ± 0.12d 73.5 ± 15.0c
E1 þ Meso1 4.25 ± 0.33b 48.0 ± 5.44b 0.95 ± 0.17c 179 ± 33.5b
E3 þ Meso3 6.19 ± 0.69a 114 ± 14.2a 1.94 ± 0.38a 469 ± 46.3a
ANOVA (One-way) <0.001*** <0.001*** <0.001*** <0.001***
ANOVA (Two-way):
Earthworm <0.001*** <0.001***
Mesofauna 0.052 <0.001***
Earthworm  Mesofauna 0.712 0.595

Levels of significance: *<0.05; **

<0.01; ***
<0.001.

Table 3
Soil chemical properties in microcosms on day 1 and 64. Values (means ± SE, n ¼ 4) followed by the same letter within a column are not statistically different at P < 0.05.

TC (%) DOC (mg kg1) TN (g kg1) NO 1

3 (mg kg ) NHþ
4 (mg kg
1
) MBC (mg kg1) MBN (mg kg1)

Day 1 7.5 ± 0.1 472 ± 16 6.1 ± 0.2 55 ± 1 11.2 ± 0.2 nd nd

Day 64
Control 7.1 ± 0.1 247 ± 5c 6.0 ± 0.1 118 ± 11c 9.6 ± 0.4d 519 ± 27c 65.1 ± 2.7d
E1 7.3 ± 0.1 346 ± 20a 5.8 ± 0.1 193 ± 15b 11.3 ± 0.1c 549 ± 13c 76.9 ± 3.8c
E3 7.4 ± 0.5 319 ± 1a 5.9 ± 0.2 283 ± 18a 27.0 ± 3.1b 604 ± 21b 86.4 ± 8.7c
Meso1 7.1 ± 0.0 360 ± 11a 6.1 ± 0.2 190 ± 35b 12.2 ± 0.9c 800 ± 76a 89.3 ± 9.1c
Meso3 7.2 ± 0.1 356 ± 8a 6.0 ± 0.2 174 ± 40b 11.5 ± 1.0c 822 ± 47a 123 ± 4.3b
E1 þ Meso1 7.2 ± 0.3 303 ± 2b 5.8 ± 0.2 208 ± 10b 11.4 ± 0.1c 643 ± 17b 321 ± 7.8a
E3 þ Meso3 7.4 ± 0.1 305 ± 9b 6.4 ± 0.1 275 ± 7a 53.9 ± 9.1a 648 ± 12b 331 ± 2.2a
ANOVA ns <0.05 ns <0.05 <0.05 <0.05 <0.05

Abbreviations: DOC, dissolved organic carbon; MBC, microbial biomass carbon; MBN, microbial biomass nitrogen; nd, not determined; ns, statistically not significant; TC, total
carbon, TN, total nitrogen. See treatment details in Table 1.
296 H. Wu et al. / Soil Biology & Biochemistry 88 (2015) 294e297

(A) effects on soil physicochemical characters (especially soil DOC, NO

3
8000
Control a and NHþ availabilities) and microbial activities (Table 3;
CO2 cumulative emissions (mg CO2-C kg soil)

4
E1 a
7000 Blagodatsky and Smith, 2012; Kuiper et al., 2013; Thakur et al.,
E3
-1

Meso1 2014).
6000 Meso3 Little is known about the interactive effects between earth-
b
E1+Meso1 worms and other soil fauna on soil gas emissions (Blouin et al.,
b
5000 E3+Meso3
2013). Our results suggested that interactions between earth-
c
4000
c
c
worm and mesofauna has no significant effect on soil CO2 and N2O
emissions, and that earthworm probably play the dominant roles in
3000 determining soil CO2 and N2O emissions where the groups coexist.
These findings might thus indicate that the belowground process
2000 ANOVA (Two-way)
***
was indeed mainly driven by species identity among soil fauna
Earthworm < 0.001
(Heemsbergen et al., 2004; Collison et al., 2013).
1000 Mesofauna 0.051
Earthworm × Mesofauna 0.450
0 Acknowledgments
0 10 20 30 40 50 60 70

This study was supported by projects from the National Natural

(B)
2100 Science Foundation of China (41171047 and 41371261), the Key
ANOVA (Two-way)
*** Research Program of the Chinese Academy of Sciences (KZZD-EW-
N2O cumulative emissions (μg N2O-N kg soil)

Earthworm < 0.001 a

1800 *** TZ-16), and the Science and Technology Development Program of
Mesofauna < 0.001
-1

Jilin Province, China (20140101004JC). We thank Dr. Darold Batzer,

Earthworm × Mesofauna 0.444 b

1500
Lynette Abbott, and two anonymous reviewers for their valuable
comments and the assistances from all staff in the Sanjiang Mire
Wetland Station.
1200

900
600
300
0 Soil Science 64, 161e182.
0 10
Fig. 1. Cumulative CO2 (A) and N2O (B) emissions over a 64-day microcosm incubation.
Significance of the different fauna and their interactions are listed (*<0.05; **<0.01;
***
<0.001). Values (means ± SE, n ¼ 3) on day 64 followed by the same letter are not
statistically different at P < 0.05. Abbreviations see treatment details in Table 1.
efflux were observed, which might be attributed to their relatively
low biomass (Filser, 2002). The 29.8e42.4% enhancement range of
N2O emissions by mesofauna (Table 2) was consistent with those
roles of mesofauna on soil N mineralization rates (Kaneda and
Kaneko, 2011).
The cumulative soil CO2 emissions from the
earthworm þ mesofauna treatments were similar with those from
earthworms alone, at the two densities (Fig. 1A). The cumulative
N2O emissions from the earthworm þ mesofauna treatments were
similar with those from earthworms alone at low densities (Fig. 1B).
At high densities, the cumulative N2O emissions from the
earthworm þ mesofauna treatments (E3 þ Meso3; 1.97 mg N kg1
DW soil) were significantly lower than the sum of earthworms
alone (E3; 1.72 mg N kg1 DW soil) and mesofauna alone (Meso3;
0.59 mg N kg1 DW soil), although slightly higher than those under
E3 (Fig. 1B). However, two-way ANOVA demonstrated no signifi-
cant interactive effects existed between earthworms and meso-
fauna on soil CO2 and N2O emissions (Table 2, Fig. 1). The
cumulative CO2 (F ¼ 22.3, P < 0.01) and N2O (F ¼ 18.9, P < 0.01)
emissions were significantly affected by earthworm density, but
not mesofauna density (Fig. 1). The differences among earthworms,
mesofauna and their combinations on soil CO2 and N2O emissions
probably resulted from their functional dissimilarities
(Heemsbergen et al., 2004; Fox et al., 2006), and their different
c
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