Bioresource Technology 132 (2013) 320326

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Chemical and spectroscopic characterization of water extractable

organic matter during vermicomposting of cattle dung
Baoyi Lv, Meiyan Xing , Jian Yang, Wensheng Qi, Yongsen Lu
Key Laboratory of Yangtze River Water Environment , Ministry of Education, State Key Laboratory of Pollution Control and Resources Reuse, College of Environmental Science
and Engineering, Tongji University, Shanghai 200092, China

h i g h l i g h t s
" Vermicomposting degraded the protein-like groups while increased the humic acid-like compounds in the WEOM.
" Fluorescence regional integration (FRI) can reveal the transformation and humication process during the vermicomposting.
" WEOM is very useful to monitor the organics transformation and assess the maturity in the vermicomposting.

a r t i c l e

i n f o

Article history:
Received 13 August 2012
Received in revised form 31 December 2012
Accepted 4 January 2013
Available online 16 January 2013
Keywords:
Vermicomposting
Water extractable organic matter (WEOM)
Fourier transform infrared (FT-IR)
Excitationemission matrix (EEM)
Fluorescence regional integration (FRI)

a b s t r a c t
This work illustrated the characteristics and transformation of water extractable organic matter (WEOM)
during vermistabilization (Eisenia fetida) of cattle dung by means of chemical and spectroscopic methods.
The independent experiment conducted in triplicate was sampled at the 0, 7, 14, 21, 35, 60 and 90 days.
Results showed that the DOC kept steady around 2.7 g kg 1 after day 60 and the DOC/DON ratio
decreased from 19.77 to 5.26 till the end of vermicomposting. On the other hand, vermicomposting
decreased the aliphatic, proteinaceous, carbohydrates components and increased the aromaticity and
oxygen-containing functional groups in the WEOM. Moreover, uorescence spectra and uorescence
regional integration (FRI) results indicated that protein-like groups were degraded and fulvic and humic
acid-like compounds were evolved during the vermicomposting process. In all, this study suggested the
suitability of WEOM for monitoring the organics transformation and assessing the maturity in the
vermicomposting.
Crown Copyright 2013 Published by Elsevier Ltd. All rights reserved.

1. Introduction
In China, the intensication of cattle breeding has resulted in an
increase in the production of biodegradable organic wastes, which
need to be efciently recycled due to the environmental problems
associated with their indiscriminate application to agricultural
elds (Bernal et al., 2009). Vermicomposting is a popular technique
characterized by the combined action of earthworms and microorganisms, thereby accelerating the stabilization of organic matter
and greatly modifying its physical and biochemical properties (Aira

Abbreviations: WEOM, water extractable organic matter; DOC, dissolved organic

carbon; DTN, dissolved total nitrogen; DON, dissolved organic nitrogen; DOC/DON,
the quotient of dissolved organic carbon and dissolved organic nitrogen; FCD, fresh
cattle dung; SUVA254, specic UV absorption at 254 nm; FRI, uorescence regional
integration; FT-IR, Fourier transform infrared; EEM, excitationemission matrix; E4/
E6, the quotient of absorbance at 465 and 665 nm; GFC, gel ltration chromatography; SFI, specic uorescence intensity.
Corresponding author. Tel./fax: +86 21 65984275.
E-mail addresses: xingmeiyan@tongji.edu.cn, xmy5000@163.com (M. Xing).

et al., 2002; Lazcano et al., 2008). This method is easy to operate,

odorless, cost effective, pathogen free and environmental friendly
(Khwairakpam and Bhargava, 2009; Lazcano et al., 2008; Li et al.,
2011). Additionally, the signicant higher nitrogen, phosphorus
and humic contents in the end products of vermicomposting can
help to improve soil fertility and stimulate plant growth (Arancon
et al., 2005).
Although earthworms accelerate the vermicomposting process
by modifying the substrate and stimulating the microbial metabolic activity (Lazcano et al., 2008), the biochemical degradation
of the organic matter is carried out by the microorganisms, either
living free and/or dwell in the earthworm gut (Aira et al., 2002;
Benitez et al., 1999). Since most of the organic matter is transformed by microorganisms in the water-soluble phase (Caricasole
et al., 2010), WEOM stands for the most active fraction of the organic waste. Moreover, WEOM was most subject to change and it
could directly reect the organic matter transformation processes
(Said-Pullicino et al., 2007; Caricasole et al., 2010). Consequently,
the composition of WEOM has been suggested as a better indicator

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B. Lv et al. / Bioresource Technology 132 (2013) 320326

of stability for the organic matter than that of the solid phase (Caricasole et al., 2010). For example, the evolution of WEOM extracted
from aerobic composting has been regarded as a good indicator of
the overall transformation and maturity of organic matter (SaidPullicino et al., 2007). Therefore, studying the changes of WEOM
is very helpful to understand the bio-stabilization process.
Previous studies on vermicomposting focused on the stabilization of various solid wastes, the application of vermicompost as a
soil organic amendment or fertilizer and the earthworm population dynamics (Garg et al., 2006; Aira et al., 2007; Renuka Gupta,
2008). In addition, the characteristics of WEOM in the initial and
nal vermicomposts of sewage sludge and cow dung were also reported (Xing et al., 2012). However, the chemical and spectroscopic
behaviors of WEOM during vermicomposting process have not
been fully understood.
WEOM consists of a heterogeneous mixture of compounds with
different molecular sizes and complexities, ranging from simple
sugars and organic acids to complex proteins and humic colloids
(Traversa et al., 2010). An integration of various techniques is a
prevalent way to characterize the features of WEOM. Many indices,
including dissolved organic carbon (DOC), specic ultra-violet
absorbance (SUVA), and uorescence excitationemission matrix
(EEM) spectroscopy etc., were used to investigate the properties
of WEOM (Shao et al., 2009; He et al., 2011a). DOC can act as a general indicator of WEOM, while SUVA254, E4/E6 and Fourier transform infrared spectra (FT-IR) can provide more detailed
information on WEOM, such as the aromaticity and humication
degree of derived compounds (Weishaar et al., 2003; Saadi et al.,
2006; He et al., 2011a). Furthermore, uorescence excitation
emission matrix (EEM) spectroscopy combining with uorescence
regional integration (FRI) technique can provide an overall view of
uorescent properties of WEOM in a selected spectral range, which
has been employed in structural identication and stability assessment of organic wastes (Zhu et al., 2011).
Keeping in light of the above facts, we hypothesized that the
WEOM could show obvious changes during the vermicomposting
process and be also very helpful to understand the stabilization
process in the vermicomposting. Therefore, the aims of this study
were to examine the chemical and spectroscopic characteristics
of WEOM at different stages during vermicomposting of cattle
dung by using various analytical approaches, as well as to investigate the transformation of WEOM and the vermicompost stability
based on the WEOM information obtained.

321

Triplicate vermireactors were established in this study. Moisture

content was maintained at 80 10% by periodic sprinkling of distilled water during the whole process. The plastic containers were
covered with perforated lids and settled in dark with temperature
kept at 23 1 C. Homogenized samples (about 50 g wet basis)
were collected at 0, 7, 14, 21, 35, 60 and 90 days of each experiment. Subsamples were air-dried, ground to pass through a 20
meshes sieve and stored in a desiccator for further analysis.
2.2. Extraction of WEOM
The extraction of WEOM from vermicompost samples was performed in a horizontal shaker with deionized water (solid to water
ratio of 1:20, w/v) for 24 h at room temperature. The suspensions
were centrifuged at 10,000g for 10 min and ltered through a
0.45-lm pore-size membrane lter (Said-Pullicino et al., 2007).
Part of the extract was freeze-dried and placed in sealed plastic
containers.
2.3. WEOM analysis
2.3.1. Organic matter contents analysis
Dissolved organic carbon (DOC) and dissolved total nitrogen
(DTN) of the extracts were measured by a TOC-VCPN analyzer (Shimadzu, Japan). The dissolved organic nitrogen (DON) content was
calculated as (DTN) (NH4+N + NO3 N). UV absorption at
254 nm of the WEOM was measured using a UV 765 spectrophotometer (Shanghai Precision & Scientic Instrument Co., Ltd.,
Shanghai, China). Before measurement, all solutions were diluted
to DOC concentrations <10 mg L 1. The specic UV absorption
(SUVA254) was determined by normalizing UV absorbance at
254 nm to the corresponding concentration of dissolved organic
carbon. The E4/E6 was given as the ratio of absorbance which measured at 465 and 665 nm.
2.3.2. FT-IR spectra
The Fourier transform infrared (FT-IR) spectra can reveal the
major functional groups of organic matters and predict the major
components (He et al., 2011a). In the present study, about 1 mg
freeze-dried WEOM sample was mixed thoroughly with 200 mg
of dried spectrometry grade KBr and pressed to a pellet under reduced pressure. The pellet was immediately measured after preparation using a Nicolet 5700 FT-IR spectrophotometer (Madison,
WI) (Xing et al., 2011). The spectra were recorded in the range of
4000400 cm 1 with a 2 cm resolution.

2. Methods
2.1. Vermicomposting process and sampling
The fresh cattle dung (FCD) was obtained from a cattle farm in
Pudong district, Shanghai, China. In order to avoid the damage of
the high moisture content and anaerobic fermentation to earthworms, the cattle dung was naturally dried under sunlight for
1 week with periodic turning over before used. Eisenia fetida maintained in the laboratory with cattle dung as culturing substrate
were randomly picked from several stock cultures. E. fetida was
chosen because it had wide tolerance of environmental variables
and had been extensively applied in the vermicomposting of organic matter such as animal manures (Khwairakpam and Bhargava,
2009; Suthar, 2009).
A lab-scale vermicomposting experiment was carried out in
plastic containers (40 cm diameter  25 cm depth) with 2 kg cattle
dung (dry basis) as substrate, followed by manually turning over
every 24 h for 7 days to eliminate toxic volatile substances. Subsequently, 250 non-clitellated earthworms (E. fetida) with an average
weight of 200250 mg per unit were inoculated into the container.

2.3.3. GFC analysis

Molecular weight distribution of the WEOM was analyzed by a
Gel Filtration Chromatography analyzer (LC-10ADVP, Shimadzu,
Japan) using TSKgel G4000PWXL column (TOSOH, Japan) equipped
with RID-10A detector on elution with aqueous Milli-Q water.
Polyethylene glycol with molecular weight (1,169,000; 771,000;
128,000; 11,840; 4020; 620 and 194 Da) was used for the calibration standards (Shao et al., 2009). 50 lL sample was injected into
the column and the ow rate was 0.5 mL min 1. The elution of elements at each time interval was collected by an automatic fraction
collector and automatically analyzed using UV spectroscopy and
dissolved organic matter analyzer (Xing et al., 2011).
2.3.4. Fluorescence spectra analysis
Fluorescence EEM spectra was recorded using an F-4600
(Hitachi, Japan) uorescence spectrophotometer in a clear quartz
cuvette. Emission and excitation slits were set at a 5-nm band
width, and a scan speed of 12,000 nm min 1 was selected. The
EEM spectra were recorded by scanning the emission wavelength
over the range 250600 nm at 2 nm increments, while the

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B. Lv et al. / Bioresource Technology 132 (2013) 320326

excitation wavelength increased gradually from 200 to 500 nm at

10 nm increments (Yu et al., 2010). The voltage of photomultiplier
tube (PMT) was set as 750 mV for low-level light detection. The
temperature of the samples was maintained at room temperature
(20 C) during the analyses and no pH adjustment was made prior
to the uorescence analysis as the pH of samples did not change
signicantly (Yu et al., 2010). The areas affected by rst- and
second-order Rayleigh and Raman scatter were adjusted by interpolation, and then the FRI technique was adopted for data analysis
(Chen et al., 2003). Surfer 8.0.1 was used to plot uorescence
spectra.
3. Results and discussion
3.1. Organic matter contents
3.1.1. DOC
DOC is the most biological and chemical active fraction in organic waste (Caricasole et al., 2010). As the DOC content of the
WEOM decreases, the substrates become more stable (He et al.,
2011b). The concentrations of DOC extracted from different stages
are shown in Table 1. In the initial 14 days, the DOC concentration
showed no signicant change, after then the DOC decreased rapidly from 4.30 to 2.70 g kg 1 until the day 60, which was possibly
attributed to the fast degradation of easily biodegradable organic
matter, especially carbohydrates, organic acids and hemicellulose
(Hsu and Lo, 1999). Moreover, it is worth noting that the earthworms in vermicomposting also assimilated a part of organics for
their growth and reproduction. Zhao et al. (2010) reported that
13.6% of the enhanced sludge stabilization was due to the direct
digestion by earthworms in vermilter designed for sewage sludge
treatment. After day 60, the DOC concentration kept steady at
around 2.7 g kg 1 which was much lower than the threshold value
of 4.0 g kg 1 suggested as the indicator of maturity (Zmora-Nahum
et al., 2005). In the present study, our result is similar to the report
of Aira et al. (2007), in which the vermicomposting of pig manure
reduced the DOC signicantly, and this would be more evident
along with the vermicomposting process. However, Lazcano et al.
(2008) reported an opposite result of minor increase of DOC after
vermicomposting of cattle manure, which was largely due to the
shorter processing time.
3.1.2. SUVA254 and E4/E6
SUVA254 has been widely used as an index for aromatic compounds within dissolved organic matter (Weishaar et al., 2003).
As shown in Table 1, the SUVA254 value increased sharply from
1.55 to 2.70 L mg 1 m 1 in the rst 35 days and then rose slightly
to 2.93 L mg 1 m 1 at the end of experiment. This is consistent
with the report of Xing et al. (2012) that SUVA254 increased greatly
after the vermicomposting treatment. The substantial increase in
SUVA254 could explain its association with the rapid transforma-

tion of non-aromatic compounds, which led to a relative enrichment in aromatics (Vinceslas-akpa and Loquet, 1997; Caricasole
et al., 2010). Furthermore, an increase of SUVA254 value suggests
a higher degree of aromaticity and molecular weight (He et al.,
2011a). Therefore, vermicomposting can increase the aromaticity
of the WEOM.
The E4/E6 ratio was calculated on the absorption intensities at
465 and 665 nm. It was negatively related to the aromatic polycondensation degree and molecular weight of humic substances (Chen
et al., 1977). Although E4/E6 ratio was not always valid in nonhumied materials, it has been used for characterizing the WEOM
from compost (Traversa et al., 2010). As shown in Table 1, the high
E4/E6 values in the middle period of the vermicomposting process
would imply the occurrence of low molecular weight aromatic
molecules, which was possibly generated by the degradation of labile organic matter. On the contrary, the low E4/E6 ratio at the end
of the experiment might indicate the high contents of large molecular weight molecules, such as humic- and fulvic-like compounds,
which were usually present in the WEOM of matured vermicomposts (Romero et al., 2007).
3.1.3. DON and DOC/DON
The DOC/DON ratio is more reliable for compost or vermicompost stability compared to the ratio of total organic carbon and
nitrogen (He et al., 2011b). As listed in Table 1, vermicomposting
led an increase of DON content and a decrease of C/N ratio of
WEOM. Specically, the DON increased obviously from 0.22 to
0.38 g kg 1 in the rst 2 weeks because of solidliquid transfer
(Said-Pullicino et al., 2007). This result is consistent with the report
of Lazcano et al. (2008) that DON increased after short-term vermicomposting of cattle dung. After that, the DON showed a slight decrease and kept stable, the consumption of amino acids and
proteins by microorganisms and earthworms could explain for this
phenomenon (Said-Pullicino et al., 2007). The DOC/DON ratio decreased during the vermicomposting process except a slight increase at day 21. As DOC was bio-oxidized to CO2 or CH4,
whereas DON was lost at a lower rate, therefore the lower DOC
to DON ratio indicated the more waste was decomposed during
composting or vermicomposting (Lazcano et al., 2008). Said-Pullicino et al. (2007) also reported that the DOC/DON ratio decreased
during composting process and reached a nal value of 6.7 when
the compost matured. According to this standard, the vermicomposted cattle dung in this study could be considered stable and
mature.
3.2. FT-IR spectra
The infrared spectra were interpreted according to the previous
studies (Ellerbrock and Kaiser, 2005; Romero et al., 2007; Li et al.,
2011). Moreover, the main absorption bands and corresponding
assignments are listed in Table 2. In general, the FT-IR spectra of
WEOM extracted from different vermicomposting stages were

Table 1
Evolution of the main parameters of water extractable organic matter (WEOM) during the vermicomposting process.
1

Days

DOC (g kg

0
7
14
21
35
60
90

4.35 0.08d
4.40 0.03d
4.30 0.06d
3.82 0.08c
3.01 0.02b
2.70 0.03a
2.53 0.01a

SUVA254

E4/E6

DON (g kg

1.55 0.11c
1.79 0.12c
2.12 0.21cb
2.56 0.12ba
2.70 0.14ba
2.89 0.06a
2.93 0.15a

9.01 0.63b
9.64 0.57b
5.45 0.51a
5.87 0.68a

0.22 0.02c
0.36 0.04ba
0.38 0.03ba
0.29 0.01cb
0.30 0.03cb
0.32 0.11cba
0.48 0.09a

Data with the same letter are not signicantly different (Tukey HSD test, a = 0.05).
Unit for DOC and DON dry weight basis.
Unit for SUVA254 L mg 1 m 1.

DOC/DON
19.77 1.25d
12.22 1.08cb
11.42 0.56cb
13.18 0.31c
10.06 0.90cb
8.43 1.01ba
5.26 0.84a

B. Lv et al. / Bioresource Technology 132 (2013) 320326

Table 2
Assignment of typical infrared bands in FT-IR spectra of organic matter.
Wave number
(cm 1)

Vibration

Functional group or component

34073384
29312925
and 2854
16551640

OH stretching
CH stretching

Phenols and carboxylic groups

Aliphatic methylene group
Amide I, carboxylates
Aromatic ring modes, alkenes
Amides II

1320

C@O stretching
C@C stretching
NH deformation and
C@N stretching
Aromatic C@C stretching
COO stretching
CN stretching
CN stretching

10801030

CO stretching

15701550
15151510
14211410

Lignin
Carboxylic acids
Amide III
Aromatic primary and secondary
amines
Polysaccharides or
polysaccharide-like substances

323

molecular weight fractions degraded quickly with time which can

be explained as the drop of fraction above 106 Da. This is in accordance with the report by Xing et al. (2012) that vermicomposting
degraded labile organic matter rapidly into smaller compounds
coupled with hydrolysis. Moreover, the molecular weight fraction
between 103 and 106 Da and below 103 Da increased in the rst
21 days. So given the DOC had no obvious variations in this period,
it indicated that the hydrolysis/transformation of the labile macromolecular organic matters mainly occurred in the preliminary
stage of the vermicomposting. On the other hand, the proportion
of fraction with molecular weight below 103 Da decreased, while
the fraction with molecular weights above 106 Da increased after
day 35. It seemed that the re-polymerization and condensation
of new macromolecules components of WEOM, like humic and fulvic substances occurred in this period (Said-Pullicino et al., 2007;
Shao et al., 2009).
3.4. EEM spectra

similar but varied signicantly in the relative intensity of absorption bands. With increasing vermicomposting time, the main
changes in FT-IR spectra were summarized as follows: (1) an obvious increase of the relative intensity at 33743407 cm 1 (OH
groups stretching); (2) a signicant decreasing trend of intensity
at 29362958 cm 1 (aliphatic CH stretching) that could be attributed to the biodegradation of lipids and carbohydrates; (3) an increase of the relative intensity at 16331643 cm 1 (aromatic
C@C, C@O stretching of amide groups, quinonic C@O and/or C@O
of H-bonded conjugated ketones); (4) a decrease of the relative
intensity at 1558 and 1510 cm 1 principally ascribed to the degradation of amide II and aromatic skeleton, like lignin; (5) a marked
increase at 13951421 cm 1 (carboxylic acids and amide III); (6) a
decrease of the intensity in the region of 10881100 cm 1 possibly
due to the decrease of polysaccharides or polysaccharide-like
substances.
In sum, these observations indicated the decrease of aliphatic
materials, N-containing groups (probably proteinaceous materials)
and carbohydrates, and the enrichment of the more oxidized and
aromatic components in the WEOM during vermicomposting of
cattle dung. Thus, the FT-IR results further conrmed and complemented the previous results that vermicomposting process caused
the disappearance of easily biodegradable compounds and enhanced the increase of aromatic compounds (Li et al., 2011; Busato
et al., 2012).
3.3. Molecular weight distribution
The molecular weight distribution of the WEOM at different
stages of vermicomposting is shown in Table 3 according to the
method proposed by Shao et al. (2009). Two main parts were found
in the initial substrates (0 day), and the fraction with molecular
weight above 106 Da accounted for the majority of the WEOM. This
was attributed to the characteristics of cattle dung, which was rich
in macromolecular organic matter, like polysaccharides. The high-

Table 3
Molecular weight distribution of WEOM at different stages of vermicomposting.
Days

0
7
14
21
35
60
90

Molecular weight (103 Da)

<1

11000

>1000

2.29
15.54
16.31
14.62
9.37
7.12
4.66

36.21
78.71
77.11
77.65
85.04
87.54
80.24

61.50
5.75
6.58
7.73
5.59
5.54
15.10

Fluorescence EEM spectra of the WEOM fractions in different

vermicomposting stages are depicted in Fig. 1. The excitation/
emission (Ex/Em) wavelength pairs and the specic intensity of
uorescence peaks are qualitatively listed in Table 4.
Two peaks were detected from the WEOM at the beginning
(0 day). Peak A was characterized by an Ex/Em wavelength pair
of 220/340 nm, which fell in the region related to the typical aromatic protein region, like tryptophan (Chen et al., 2003). Peak B
represented an Ex/Em wavelength pair of 280/328 nm and associated with the soluble microbial byproduct-like materials (Chen
et al., 2003). During vermicomposting, the SFI of Peaks A and B presented a trend of decrease in spite a slightly rose at day 35. Both
peaks nally disappeared at the end of the experiment, which
could be explained as the bio-degradation of organic matters as
the result of microbial and earthworm activities (Marhuenda-Egea
et al., 2007). This result is consistent with that reported by Xing
et al. (2012).
At day 7, the third peak (Peak C) emerged at an Ex/Em wavelength pairs of 270/436 nm, which was associated with fulvic
acid-like substances (Chen et al., 2003). The presence of this peak
indicated the formation of fulvic acids. Marhuenda-Egea et al.
(2007) reported that different molecular compounds derived from
lignin and other degraded plant materials could contribute to the
uorescence of fulvic acids. At the same time, another fulvic
acid-like compounds peak (Peak D) with an Ex/Em wavelength pair
of 230/412 nm was also detected. This peak was used to monitor
changes in fulvic acid-like materials during the composting of
MSW (Said-Pullicino et al., 2007; He et al., 2011a). However, as
the vermicomposting proceeded, Peak D disappeared after day
21. According to Marhuenda-Egea et al. (2007), the fulvic acid-like
components can be synthesized into the humic macromolecules
during their formations.
The last peak (Peak E) was observed at day 60 with an Ex/Em
wavelength pair of 320/416 nm, and fell into the region dened
as a longer excitation wavelength (>280 nm) and a longer emission
wavelength (>380 nm) which was related to humic acid-like
organics (AntZar-Ladislao et al., 2006). Its intensity increased
gradually as vermicomposting progressed. Provenzano et al.
(2001) reported peaks characterized by an Ex/Em wavelength pair
of 330/425 related to humic acids in domestic solid waste after
30 days of composting treatment. Furthermore, He et al. (2011a)
also reported peaks centered at Ex/Em wavelength pair of 313/
406 related to humic acid in WEOM which was extracted from
composted municipal solid waste. In the current study, this peak
emerged when the DOC and SUVA254 became steady, which implied a close relation with the maturity of vermicomposted cattle
dung. Additionally, the increase of the peak intensity indicated a

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B. Lv et al. / Bioresource Technology 132 (2013) 320326

500

500

300

480

0d

450

440

450

280

7d

260
240

400

220

360

400

400

200

280

350

300

Peak B

240

Ex (nm)

Ex (nm)

320

180

Peak C

350

160
140

200

Peak A

120

300

160

100

Peak D

80

120

250

60

250

80

40

40

200

250

20

300

350

400

450

500

550

200
250

600

300

350

Em (nm)

450

500

550

600

Em (nm)

500

450

400

500

300
280

14 d

260

450

300
280

21 d

260
240

240

220

220

400

200
180
160

350

140

Ex (nm)

Ex (nm)

400

200
180
160

350

140
120

120

300

300

100

100
80

80
60

250

60

250

40

40
20

20

200
250

200
250

300

350

400

450

500

550

600

300

350

Em (nm)

400

450

500

550

600

Em (nm)

500

500

300

35 d
450

300

60 d

280

280

450

260

260
240

240

400

400

200
180
160

350

140

Ex (nm)

Ex (nm)

220

220

200

Peak E

180
160

350

140
120

120

300

100

300

100
80

80
60

250

40

60

250

40
20

20

200
250

300

350

400

450

500

550

600

Em (nm)

200
250

300

350

400

450

500

550

600

Em (nm)

500

300

90 d

280
260

450

240
220

Ex (nm)

400

200
180
160

350

140
120

300

100
80
60

250

40
20

200
250

300

350

400

450

500

550

600

Em (nm)

Fig. 1. Fluorescence excitation-emission matrix spectra of WEOM at different stages of vermicomposting. Note: normalized to 1 mg L

DOC by dividing corresponding DOC.

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B. Lv et al. / Bioresource Technology 132 (2013) 320326

Table 4
Ex/Em maxima of WEOM during vermicomposting.
Days

Peak A
a

0
7
14
21
35
60
90
a
b

Peak B
b

Peak C

Peak D

Peak E

Ex/Em

SFI

Ex/Em

SFI

Ex/Em

SFI

Ex/Em

SFI

Ex/Em

SFI

220/342
220/340
220/340
220/342
220/340
230/342

413.5
156.5
172.7
169.2
198.1
87.5

280/330
280/328
280/328
280/330
280/320
280/336

430.4
170.7
186.2
163.6
203.1
138.1

270/436
270/436
260/436
270/440
260/442
270/446

108.1
126.2
176.5
205.6
210.6
228.5

230/422
230/422

105.5
107.9

320/416
320/420

159.8
173.3

Ex/Em referred to the excitation/emission wavelength pairs.

SFI referred to the specic uorescence intensity.

WEOM was increased throughout the vermicomposting. Furthermore, the transformation of WEOM during vermicomposting was
characterized by the decrease of tyrosine- and tryptophan-like
materials and the increase in humic and fulvic-like substances,
and nally resulted in the enhanced stability of the cattle dung.
3.5. Implications for vermicomposting

Fig. 2. Evolution of Pi,n values of the ve regions during vermicomposting.

higher degree of humication or maturation along with the vermicomposting process.

Fluorescence regional integration (FRI) is a new analytical approach, which can analyze all the wavelength-dependent uorescence intensity data from EEM spectra (Chen et al., 2003; Zhu
et al., 2011). This technique can be used to capture the heterogeneity of WEOM (Shao et al., 2009). Thus, to better understand the
transformation of WEOM, the FRI was employed to analyze
the EEM data in this study. As described by Chen et al. (2003),
the EEM spectra was divided into ve excitation-emission regions
(region I: tyrosine-like organic compounds; region II: tryptophanlike organic compounds; region III: fulvic acid-like materials;
region IV: soluble microbial byproduct-like materials; region V:
humic acid-like materials). The percent uorescence responses
(Pi,n) was determined by normalizing the cumulative excitationemission area volumes to relative regional areas.
The Pi,n of the ve regions are displayed in Fig. 2 as a function of
the vermicomposting time. On the one hand, the Pi,n of the regions I
and II exhibited a decrease trend during the vermicomposting process, which indicated the decrease of simple aromatic proteins,
such as tyrosine-like and tryptophan-like organic compounds. On
the other hand, the Pi,n of regions III and V increased as the vermicomposting processed, revealing an increase of humic-like and fulvic-like materials in the WEOM (Marhuenda-Egea et al., 2007).
Similar result was found by Marhuenda-Egea et al. (2007) during
composting of winery and distillery residues. Moreover, the PV,n/
PIII,n ratio showed no obvious change in the rst 21 days while it
rose sharply after that, which indicated the formation of humic
substances (Zhu et al., 2011). Busato et al. (2012) also reported
the similar phenomenon that humic acids increased greatly from
day 30 during maturation of vermicompost. As described by He
et al. (2011a), the greater humication degree was associated with
more complex and condensed aromatic structure, which were
resistant to bio-degradation. Therefore, the stabilization degree of

Composting and vermicomposting are two of the best-known

processes for bio-stabilization of solid organic wastes (Lazcano
et al., 2008). As one of the research hot spots, WEOM has been
studied by many researchers in composting process. It has been
conrmed not only crucial to understand the transformation of organic substrate (Marhuenda-Egea et al., 2007; Caricasole et al.,
2010), but also very helpful to assess the maturity of composted
substrates (Shao et al., 2009; Zhu et al., 2011). In this study, we
found that WEOM had also changed dramatically during the vermicomposting process. Vermicomposting decreased the DOC and
increased the SUVA254 of the WEOM, moreover, the data of FT-IR
and EEM also showed obvious changes during vermicomposting
process. These results demonstrated the vermicomposting was
characterized by the degradation of liable organic matter and the
formation of humic and fulvic-like materials in the WEOM. Furthermore, the above information based on WEOM was in accordance with the existing conclusions which obtained from humic
fraction or 13C NMR (Vinceslas-akpa and Loquet, 1997; Li et al.,
2011). As we all know, it is time-consuming and inefcient to evaluate the maturity of vermicompost via insoluble organic chemical
or biological parameters (Benitez et al., 1999; Lazcano et al., 2008),
while the analysis of WEOM, especially by means of excitation
emission matrix spectroscopy can be effective in assessing the
chemical stability and biological maturity in vermicomposting
due to its convenient operation and high sensitivity (Yu et al.,
2010). In all, the characterization and analysis of WEOM could play
an important role in monitoring the organics transformation and
evaluating the vermicomposting process.
4. Conclusions
WEOM extracted from different vermicomposting stages were
characterized by chemical and spectroscopic techniques. The
DOC concentration kept steady around 2.7 g kg 1 after day 60.
Moreover, the aromaticity of WEOM signicantly increased and
the fraction with molecular weight between 103 and 106 Da became the main part of WEOM. Vermicomposting decreased the aliphatics, alcohols, protein-like materials and polysaccharides,
whereas increased the humic-like and fulvic-like materials in the
WEOM. These ndings revealed that vermicomposting drove the
transformation of labile WEOM into stable substances. In conclusion, WEOM can be regarded as a good indicator to assess the
maturity during vermicomposting.

326

B. Lv et al. / Bioresource Technology 132 (2013) 320326

Acknowledgements
The research was funded by the National Natural Science Foundation of China (NSFC, No:51109161), the PhD Programs Foundation of Ministry of Education of China (20110072120029), the
Fundamental Research Funds for The Central Universities
(0400219187), the Open Analysis Fund for Large Apparatus and
Equipments of Tongji University (No. 2012055), the National Spark
Program of China (2010GA680004). The author would like to thank
Shaobo Liang (University of Idaho) for reading through the manuscript and providing suggestions. Thank the editor and anonymous
referees to improve the quality of the manuscript.

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