Bioresource Technology 116 (2012) 485491

Contents lists available at SciVerse ScienceDirect

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

Rapid production of maggots as feed supplement and organic fertilizer

by the two-stage composting of pig manure
Feng-Xiang Zhu a,b, Wei-Ping Wang b, Chun-Lai Hong b, Ming-Guang Feng a,, Zhi-Yong Xue b,
Xiao-Yang Chen b, Yan-Lai Yao b, Man Yu b
a
b

Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou 310058, PR China
Institute of Environment, Resource, Soil and Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, PR China

a r t i c l e

i n f o

Article history:
Received 16 February 2012
Received in revised form 3 April 2012
Accepted 3 April 2012
Available online 9 April 2012
Keywords:
Pig manure
Two-stage composting
Maggot production
Fertilizer production

a b s t r a c t
A two-stage composting experiment was performed to utilize pig manure for producing maggots as feed
supplement and organic fertilizer. Seven-day composting of 1.8 ton fresh manure inoculated with 9 kg
mixture of housey neonates and wheat bran produced 193 kg aging maggots, followed by 12 week composting to maturity. Reaching the thermophilic phase and nal maturity faster was characteristic of the
maggot-treated compost compared with the same-size natural compost. Upon the transit of the maggottreated compost to the second stage, the composting temperature maintained around 55 C for 9 days
and the moisture decreased to 40%. Moreover, higher pH, faster detoxication and different activity patterns for some microbial enzymes were observed. There was a strong material loss (35% water-soluble
carbon and 16% total nitrogen) caused by the maggot culture in the rst stage. Our results highlight a
higher economic value of pig manure achieved through the two-stage composting without bulking
agents.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
In the past two decades, a burst of pig industry in China has
brought about a huge amount of manure, which is not only a reservoir of pathogens, parasites and weed seeds (Larney and Hao,
2007) but a favorable breeding substrate for the housey Musca
domestica (Farkas et al., 1998). The harmful agents must be eliminated from the manure through proper composting for the production of organic fertilizer. This is often challenged by the high water
content of 7080% in fresh pig manure because the excessive moisture is a barrier to reaching a high temperature in a manure compost. The moisture suitable for compositing often ranges from 50%
to 60%, beyond which oxygen movement is inhibited in the composite (Das and Keener, 1997; Gajalakshmi and Abbasi, 2008).
Upon eld application of immature compost, inadequate oxygen
in soil may inhibit seed germination or suppress root and plant
growth (Brinton and Evans, 2001; Said-Pullicino et al., 2007). For
this reason, bulking agents, such as sawdust and rice chaff, are usually added to the manure compost for moisture reduction. However, such bulking agents have become increasingly expensive for
composting as they are more used as alternative energy resources.
Thus, it is necessary to explore alternative means to composting
pig manure for fertilizer production.
Corresponding author. Tel./fax: +86 57188206178.
E-mail address: mgfeng@zju.edu.cn (M.-G. Feng).
0960-8524/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.04.008

On the other hand, the rapid development of aquaculture in

China needs more sources of feed proteins. The housey larvae,
namely maggots, are an ideal supply of aquaculture feeds due to
>50% crude proteins in dry weight (Akpodiete et al., 1997; Iniguezcovarrubias et al., 1994) and an excellent source of limiting amino
acids, such as lysine, methionine and phenylalanine (Ocio and
Vinaras, 1979). The maggot protein content is even higher than
those in soybean, meat and bone scrap (Iniguez-covarrubias
et al., 1994). Moreover, living maggots provide a variety of biologically active substances, such as antimicrobial peptides, lectins and
chitins (Fu et al., 2009; Hou et al., 2007). Maggot proteins added to
animal feed may stimulate animal appetite. For instance, rats consumed more maggot protein-inclusive meal (Iniguez-covarrubias
et al., 1994). Housey larvae and pupae cultivated in manure
showed a nutritive value similar to that of sh meal or animal proteins (Akpodiete et al., 1997; Miller et al., 1974; Ocio and Vinaras,
1979). Interestingly, poultry manure treated by breeding maggots
became somewhat granular for an ease of drying due to improved
aeration (Miller et al., 1974). Recently, chitosan, a maggot extract,
was even used in cosmetics and medicines (Ai et al., 2008; Jing
et al., 2007). These studies indicate that, as a source of feed proteins and biologically active substances, the manure-cultured maggots are of high economic value although exposure to the maggots
may lead to health problems in humans and animals and means of
preventing human and animal exposure will have to be developed
for the application of the maggot culture technique.

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F.-X. Zhu et al. / Bioresource Technology 116 (2012) 485491

This study sought to develop a two-stage composting technology for the use of pig manure in the production of maggots as feed
supplement and organic fertilizer. Our goal was to maximize maggot production in the rst stage and accelerate the composting to
maturity in the second stage by monitoring changes in temperature, moisture, pH, phytotoxicity and microbial enzyme activities.
2. Methods
2.1. Selecting a maggot inoculum level for composting
A triplicate experiment was conducted to determine a proper
level of maggot inoculum for the composting of fresh pig manure
at Deqing Pig Farm (Hangzhou, Zhejiang, China). Batches of
200 kg manure were atly piled to 7 cm thickness in cement
trays (2.4  2.3  0.2 m) under greenhouse conditions and inoculated with maggot inoculum at the weight ratios of 0.25%, 0.5%,
0.75%, 1.0% and 1.5%, respectively, forming ve treatments plus
blank control (not inoculated). Each gram of the inoculum consisted of 2000 neonate maggots (<24 h after hatch) and wheat
bran used for egg hatch. After 7 day composting under ambient
conditions, all aging maggots were harvested from the manure of
each tray by means of their photophobotaxis and weighted for total biomass (fresh weight). Meanwhile, water content in the treated manure was assessed with three samples from each tray. The
inoculum level leading to maximal maggot biomass and less than
65% water content was chosen for the composting experiment as
follows.
2.2. Composting experiment
A two-stage composting experiment was performed to produce
both maggots and organic fertilizer with fresh pig manure. The
rst-stage composting began from introducing 0.5% (w/w) maggot
inoculum (the optimized level) to 1.8 ton manure in three large cement trays (7.3  2.3  0.2 m), in which the manure was atly
piled as in the previous experiment. After maggot harvest on day
7, all the manure collected from the trays was piled up into a
peak-shaped compost (4.5 m in length, 2.2 m in width, and 0.8 m
in height), namely maggot-treated compost (MC), on the ground
in a rainproof workshop without walls. Besides MC, a natural compost (NC) was constructed as a control in the same way using
1.8 ton manure composted for 7 days in other three trays without
maggot inoculum. Both MC and NC were covered with plastic lm
for 24 h to stie possible residue maggots and then entered the
second stage of composting for 12 weeks, during which both composts were turned upside down every 3 days.
Three 1 kg samples were randomly taken from each tray daily
during the rst stage and from MC or NC 40 cm below the surface
at the intervals of 36 days during the second stage, respectively.
All samples were preserved at 10 C for the following analysis.
2.3. Assays for physical and chemical properties
During the composting, temperature was monitored daily by
reading 25 or 50 cm tubular mercurial thermometers (Kal Longda
Instrument Co., Tianjin, China) inserted into three sites of each tray
4 cm or each compost 30 cm below the surface. Percent water
content in each sample was assessed by drying 100 g subsamples
at 105 C for 24 h. The pH values in 10% (w/v) sample suspensions
were assessed using an electronic pH meter (Mettler-Toledo
Instruments (Shanghai) Co., Ltd., Shanghai, China). The methods
of Kjeldahl (Bremner and Mulvaney, 1982) and K2Cr2O3 oxidation
(Fan, 2007) were adopted to assay the contents (g/kg) of total
nitrogen and water-soluble carbon in each of three 5 g subsamples,

respectively. All the assays included blank controls to exclude

background effects.
2.4. Assays for microbial enzyme activities
All the samples were analyzed in triplicate assays for the overall
activities of selected microbial enzymes using previous methods
(Li et al., 2008). Sample-free blank controls were included in all
the enzyme assays to exclude background effects.
Catalase activity was assayed by shaking 100 mg wet sample in
210 ml of 14 mM H2O2 at 120 rpm for 30 min reaction at room
temperature. The reaction was terminated by adding 30 ml of
1.5 M sulfate, followed by ltration through qualitative lter paper
with medium retention and ow rate (the same below). The ltrate
of the reacted suspension was diluted to 250 ml with sterile water
(the same below). Every 25 ml of the diluted ltrate was titrated to
light red with 2 mM KMO4. The catalase activity was assessed as
the volume (ml) of 2 mM KMO4 consumed per gram of dry sample
during the 30 min reaction (Piotrowska-Cyplik et al., 2009).
The assay for the activity of alkaline phosphatase started from
treating 500 mg sample with 1.5 ml toluene for 15 min. The treated
sample was mixed with 10 ml of 26 mM benzene sodium dihydrogen phosphate (C6H5PO4Na2) and 10 ml of 0.2 mM borate buffer
(pH 10). The mixture was incubated at 37 C for 3 h reaction and
then diluted to 100 ml with water heated to 38 C. After ltration,
every 10 ml of the ltrate was mixed with 5 ml of 0.2 mM borate
buffer (pH 9.6) and diluted to 25 ml with water, followed by adding
1 ml of 6.7 mM Gibbs reagents for 20 min incubation. The mixture
was further diluted to 50 ml with water and measured for the
absorbance at 578 nm in Spectronic GENESYS 5 UVVisible Spectrophotometer (Thermo Electron Corporation, Waltham, MA,
USA). The enzyme activity was expressed as the amount (lg) of
phenol produced per gram of dry sample during the 3 h reaction.
To assay urease activity, 200 mg wet sample pre-treated with
2 ml toluene was suspended in 20 ml of 0.96 M citrate buffer (pH
6.7) and 20 ml of 2.2 M urea for 3 h incubation at 38 C. The suspension was diluted to 100 ml with warmed (38 C) water for ltration. Every 10 ml of the ltrate was supplied with 4 ml 1.33 M
phenol sodium, 3 ml 0.9% active chlorine and 10 ml water, followed by 20 min incubation at room temperature. Upon dilution
to 50 ml with water, the solution was measured for the absorbance
at 578 nm. The urease activity was assessed as the amount (lg) of
NH
4 -N produced per gram of dry sample during the 3 h reaction.
The dehydrogenase activity was assayed by suspending every
2 g sample in 5 ml of 15 mM triphenyltetrazolium chloride (TTC)
and incubating the suspension for 6 h at 30 C in dark. The suspension was mixed with 80 ml methanol (chemical purity) by shaking
for 1 h at 200 rpm. The ltrate of the mixture was diluted to 100 ml
with methanol and measured for the absorbance at 485 nm. The
enzyme activity was measured as the amount (lg) of triphenyl formazance (TPF) produced per gram of dry sample during the 6 h
reaction.
The polyphenoloxidase activity was assayed by suspending
each of 100 mg samples in 20 ml of 80 mM pyrogallol for 3 h reaction at 30 C. A mixture of the reacted suspension with 8 ml buffer
(53 mM citrate and 94 mM Na2HPO4) was repeatedly extracted
with 20 ml ether and the extract was re-suspended in 50 ml ether.
Every 5 ml of the suspension was diluted to 25 ml, followed by
measuring immediately the absorbance at 430 nm. The enzyme
activity was quantied as the amount (mg) of purpurogallin
(PPG) produced per gram of dry sample during the 3 h reaction
(Ma et al., 2003).
The assay of nitrate reductase activity started from suspending
each of 200 mg samples in 2 ml of 0.1 M KNO3 and 0.4 mM CaCO3.
A mixture of the suspension with 2 ml of 1% glucose was vacuumdried for 3 min and then incubated for 24 h at 30 C. The product

F.-X. Zhu et al. / Bioresource Technology 116 (2012) 485491

was suspended in 25 ml water plus 2 ml of saturated KAl(SO4)2

solution. The resultant ltrate was diluted to 50 ml with water
and every 10 ml dilution was dried up in heated oil bath. The residue was dissolved in 15 ml water plus 2 ml of 1.6 M phenol disulfonic acid and the solution was adjusted to pH 9.5, followed by
diluting the total volume to 50 ml and reading the absorbance at
410 nm. The enzyme activity was assessed as the amount (lg) of
NO3-N reduced per gram of dry sample during the 24 h reaction.
In the assay of nitrite reductase activity, each 200 mg sample
was subjected to the same treatment as above except the use of
73 mM NaNO2 and 0.2 mM CaCO3. The diluted ltrate of 1 ml
was mixed with 4 ml reagent (2 ml of 7 mM a-naphthylamine
and 2 ml of 29 mM p-aminobenzene sulfonic acid) and 5 ml water
and the mixture was diluted to 50 ml with water for measuring the
absorbance at 525 nm. The enzyme activity was assessed as the
amount (lg) of NO2-N reduced per gram of dry sample during
the 24 h reaction.
To assay the invertase activity, each 500 mg sample was suspended in 20 ml buffer (61 mM KH2PO4 and 5 mM K2HPO4, pH
5.8) and 20 ml 20% sucrose. The suspension was mixed with
1.5 ml toluene for 15 min incubation at room temperature, followed by 23 h incubation at 38 C. The mixture was diluted to
100 ml with warmed water for 1 h incubation at 38 C. The reacted
mixture of 20 ml was blended with 20 ml Fehling solution and
20 ml water, boiled in water bath for 10 min, and then cooled to
25 C in ice bath. The cooled mixture was titrated to blue with
0.1 M Na2S2O3 after adding 8 ml of 4.6 M H2SO4, 6 ml of 3 M KI
and 0.5 ml of 0.3% starch (as indicator). The invertase activity
was measured as the amount (ml) of 0.1 M Na2S2O3 consumed
per gram of dry sample during the 24 h reaction.
2.5. Assays for phytotoxicity in samples
Two germination experiments were carried out to assay the
toxicity of the samples to the seeds of Chinese cabbage and cucumber, respectively. Aqueous extracts were prepared by shaking 10%
(w/v) sample suspensions for 30 min at 120 rpm, followed by
30 min standing and 15 min centrifugation at 4200 rpm. The aliquot of 5 ml each supernatant was poured into Petri dish (9 cm
diameter) with a lter paper, on which 20 seeds were evenly distributed in advance. The seeds were incubated for 2 days at 25 C
in dark, followed by assessing percent germination and mean root
length in each dish. The same volume of distilled water was used as
control for the seed germination. The ratios of percent germination
and mean root length in each extract-inclusive dish over those in
the control were dened as relative germination rate (%) and relative root growth (%), respectively. The product of the two ratios divided by 100 was dened as germination index (Tiquia and Tam,
1998) for the phytotoxicity of a given sample because such an index can reect the overall maturity of composting materials (Smith
and Hughes, 2004).
3. Results and discussion
3.1. Maggot production during the rst-stage composting
The rst experiment of 0.53.0 kg neonate maggot inoculum in
200 kg aliquots of fresh pig manure in cement trays resulted in signicant different maggot yields (F4,10 = 5.3, P = 0.0148) and moisture contents (F5,12 = 25.4, P < 0.0001) in the treatments of 7 day
maggot cultures. A maximal yield of 23 1.7 kg aging maggots
(near pupation) per tray (Fig. 1a) was harvested from the treatment of 1 kg inoculum (i.e., 0.5% of fresh manure weight), which
was accompanied by moisture reduction to 56.2 1.2% (Fig. 1b).
This manure moisture was well in agreement with the optima of

487

Fig. 1. Comparison of maggot yields (a) and moisture contents (b) among the
treatments of fresh pig manure at the end of the rst-stage composting. The
treatments consisted of different levels (0.251.5%) of wheat bran-inclusive
neonate maggot inoculum plus control. Different lowercase letters on the bars of
each graph denote signicant differences (Tukeys HSD, P < 0.05). Error bars: SD of
the mean from three replicates.

5060% for manure composting to maturity (Das and Keener,

1997; Gajalakshmi and Abbasi, 2008). The body weights of harvested maggots were averagely 919.3 mg per capita, decreasing
with the inoculum level.
In the enlarged two-stage experiment, the rst 7 day composting of 1.8 ton fresh pig manure inoculated at the optimized level of
0.5% maggot inoculum resulted in a harvest of 193 kg aging maggots. This maggot yield was equivalent to 10.7% of the fresh manure weight and close to the percent yield of 11.5% at the same
inoculum level in the previous experiment. The results indicated
that fresh pig manure was an excellent substrate for easy use in
the rapid production of maggots as feed supplement.
3.2. Trends of physical and chemical parameters during composting
After the rst-stage composting for maggot culture, the manure
became granular, well in agreement with the previous report on
texture of poultry manure treated by maggots (Miller et al.,
1974). The granular manure was piled up on the ground for entry
into the second-stage composting under rainproof conditions. The
two-stage composting of 13 weeks experienced roughly maggot
treating (days 19), thermophilic (days 1018), mesophilic (days
1943), and thermostable (days 4492) phases, respectively.
During the composting period, ambient mean temperature uctuated daily from 19.7 to 34.9 C (average 28 C). Under such conditions, the maggot treatment increased rapidly the composting
temperature to 43 C on day 6 and maintained 5357 C (average
54.8 C) during days 1018 (Fig. 1a). The mean temperature during
the rst two phases reached 46.9 C in MC but was only 34.5 C in
NC (paired Students t14 = 6.3, P < 0.0001). Note that the temperatures on days 79 were not measured due to the transit from the
rst stage of maggot treatment in cement trays to the second stage
of natural composting on the ground. In the last two phases, contrastingly, the temperature trend of 40.064.3 C (average

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F.-X. Zhu et al. / Bioresource Technology 116 (2012) 485491

Fig. 2. Changes of physical and chemical parameters in pig manure composts during two-stage composting. s, Maggot-treated compost. 4, Natural compost (control). (a)
Composting temperature (d, ambient mean with minimum and maximum). (b) Moisture. (c) pH. (d) Water-soluble carbon content. (e) Total nitrogen content. Error bars: SD
of the mean from three replicates.

50.8 C) in NC was signicantly higher than the trend of 36.0

50.7 C (average 41.5 C) in MC (paired Students t66 = 19.4,
P < 0.0001). These data indicated that the maggot treatment
greatly facilitated the temperature increase during the rst
18 days.
Accompanied with the temperature changes, the two manure
composts showed different moisture and pH changes during the
composting. The initial moisture of 72.3% decreased to 58% and
64% on day 6 in MC and NC, respectively. Upon the transit to the
second phase, the moisture became signicantly lower in MC than
in NC (paired Students t24 = 5.9, P < 0.0001) until it declined to
20% in both composts (Fig. 2b). The pH trend (average 8.1) in
MC was higher than that (average 7.7) in NC (paired Students
t31 = 5.3, P < 0.0001) during the period (Fig. 2c). Apparently, the

maggot treatment accelerated both moisture reduction and pH elevation from the initial levels of 72.3% and pH 6.94 than the control,
respectively.
Apart from the changes in temperature, moisture and pH during
the composting, the contents (range and mean) of water-soluble
carbon (Fig. 2d) and total nitrogen (Fig. 2e) fell in the ranges of
2.054.75 (2.88) g/kg and of 29.362.3 (36.9) g/kg in MC, respectively. These were signicantly lower than the trends of 4.07
5.08 (4.60) g/kg (paired Students t19 = 8.7, P < 0.0001) and 33.3
62.3 (44.3) g/kg measured from NC (paired Student0 s t19 = 7.4,
P = 0.018), respectively. The MC samples taken on day 10 lost
44% water-soluble carbon and 34% total nitrogen compared with
the NC counterparts. At the end of composting, the carbon and
nitrogen losses diminished to 35% and 16%, respectively. The lost

F.-X. Zhu et al. / Bioresource Technology 116 (2012) 485491

carbon and nitrogen in MC apparently contributed to the development of maggots and the release of more CO2 and ammonia during
the early stage. From the point of economic view, the cost is worth
because the harvested maggots can be sold as aquaculture feeds at
the current price of 64 US dollars per 100 kg while each ton of mature compost at the sale price of 95126 US dollars is usually converted from 3.5 tons of fresh pig manure. Noticeably, the lower
contents of water-soluble carbon in MC indicated that the maggot
treatment could reduce excessive carbonation for less CO2 release
after the early stage although the measurements were consistently
above 2%, which was higher than the level of <1.7% suggested for a
mature compost (Zmora-Nahum et al., 2005).
Overall, the maggot treatment greatly shortened the period of
composting to maturity by altering the physical and chemical
parameters. The changed parameters allowed for the recognition
of thermophilic, mesophilic and stable phases in the process of
composting, as shown previously in the composting of animal
manure or agro-industrial wastes (Bao et al., 2010; Bernal et al.,
2009; Raj and Antil, 2011). Since a composting temperature
exceeding 55 C for more than 3 days (Yu et al., 2007) or 50 C
for consecutive 8 days (Bao et al., 2010; China National Standard
for Manure Composting: GB 7959-87) was recommended to disinfect animal and plant pathogens in waste materials, the secondphase mean temperature of 54.8 C for consecutive 9 days was sufcient for pathogen elimination from MC.
3.3. Activity patterns of different enzymes during composting
The assayed enzymes in the MC and NC samples taken during
the 13 week composting included catalases to reduce the accumulation of reactive oxygen radicals, alkaline phosphatases to catalyze
the hydrolysis of organic phosphorus to inorganic forms (Raut et al.,
2008), polyphenoloxidases relating to the level of humication (Ma
et al., 2003), dehydrogenases to participate in microbial respiratory
chains (Barrena et al., 2008), ureases involved in the peptide bond
hydrolysis of organic matter and the decomposition of urea into
ammonia and carbon dioxide, nitrate and nitrite reductases involved in the transformation of nitrogen compounds, and invertases relating to the levels of humus and water-soluble organic
matter. Their activity trends are illustrated in Fig. 3.
The MC catalase activity was stabilized earlier during the rst
3 weeks (Fig. 3a) and was consistently lower than the NC counterpart from then on (Students t20 = 5.2, P < 0.0001). The polyphenoloxidase activity increased from the initial level of 107 mg PPG g1
3 h1 to a maximum in MC faster than in NC (Fig. 3b) during the
early stage and uctuated around the initial level in both composts
(Students t22 = 0.52, P = 0.61) after a rapid decrease. The dehydrogenase activities in MC and NC dropped drastically from the initial
level of 3712 lg TPF g1 6 h1 to low levels in the rst phase
(Fig. 3c), followed by uctuating similarly around the low levels
(Students t38 = 0.52, P = 0.60). Such trends are well in accordance
with the previous trends of dehydrogenase activities in the composts of municipal solid waste and sewage sludge (Wong and Fang,
2000; Vargas-Garcia et al., 2010). In an overview to the enzymes of
three types associated with redox balance and microbial activity,
their activities were stabilized more rapidly in MC than in NC,
mostly in agreement with the changes of composting temperature
and moisture in both composts. Particularly, the catalase activity
trends of MC during days 539 and of NC during days 1370 reected temperature changes in the thermophilic and mesophilic
phases in this study, as shown in the same phases of other composts (Piotrowska-Cyplik et al., 2009; Volchatova et al., 2002),
and thus could be more indicative of the compost maturity and
microbial activity.
As enzymes involved in phosphorus cycle, alkaline phosphatases were highly active in the initial samples (up to 6404 lg

489

phenol g1 3 h1), an indication for a large amount of organic

phosphate compounds existing in fresh manure (Raut et al.,
2008). Their activities in MC and NC decreased to a bottom on days
6 and 22, respectively, followed by a rebound to higher levels within one week (Fig. 3d). The subsequent activity trend was lower in
MC than in NC (Students t14 = 2.6, P = 0.02) despite a similar level
at the end of composting. Again, the phosphatase activity in MC
was stabilized more rapidly.
Among the enzymes involved in nitrogen cycles, nitrate and nitrite reductases were not active in fresh manure samples due to
their undetectable activities at the beginning time. The nitrate
reductase activity (lg NO3-N g1 24 h1) in MC was not measurable until a maximum of 5995 was observed on day 3, followed
by dropping to an undetectable level on day 16 and rebounding
to high levels from day 34 onwards (Fig. 3e). The same enzyme
activity in NC was measured as 2571798 during days 15 but became undetectable during days 613, followed by a fast rebound to
5669 on day 28 and a high-level uctuation afterward. The nitrite
reductase activity (lg NO2-N g1 24 h1) in MC was not detectable
during the 13 week composting except the periods of days 25
(115576) and days 4292 (10542720) while the same enzyme
activity in NC was not measurable until day 25 (404), followed
by an increase to a maximum of 4661 on day 70 (Fig. 3f). Interestingly, the urease activity (lg NH4-N g1 3 h1) in MC increased
from the initial level of 46629548 on day 1 but became consistently undetectable from day 3 onwards (Fig. 3g) whereas three
activity peaks were observed in NC during the periods of days 1
5 (6005804), days 2845 (1964287) and days 7282 (196
903), respectively.
Finally, the invertase activity (ml 0.1 M Na2S2O3 g1 24 h1) in
MC decreased faster to a bottom from the initial level of 297 than
that in NC during the rst 5 weeks (Fig. 3h) and then uctuated at
lower levels (Students t14 = 5.8, P < 0.0001). Our observations are
in agreement with the invertase activity trends from the beginning
time to mature stage in the composts of other materials (Ni and
Xue, 2005; Volchatova et al., 2002; Zameer et al., 2010).
3.4. Changes in the phytotoxicity of samples during composting
In the germination experiments, Chinese cabbage seeds were
more sensitive to the sample extracts, irrespective of MC or NC,
than cucumber seeds, as reported previously (Tiquia et al., 1996).
The cabbage seeds failed to germinate in the extracts of MC and
NC samples taken before days 10 and 25 (Fig. 4a), respectively,
an indication for the existence of substantial phytotoxicity in the
early samples of both composts. Subsequently, the cabbage germination index increased more rapidly to >100% in the extracts of MC
(on day 34) than NC (on day 55). Similarly, the cucumber germination index in the extracts of early NC samples maintained at the
low level of <10% for 19 days longer than that in the MC counterparts, followed by a rapid increase (Fig. 4b). Since the germination
index above 60% is indicative of the maturity of composting (Zucconi et al., 1985), our observations indicate that MC was mature for
the germination of cucumber seeds (71.4%) on day 10 and of cabbage seeds (86.5%) on day 25 whereas the NC samples showed
toxic effect on the germination of cabbage and cucumber seeds
until day 42, when both germination indices exceeded 60%. Apparently, the maggot treatment accelerated greatly the detoxication
of pig manure during the period of composting.
4. Conclusion
The two-stage composting of fresh pig manure was efcient
for the production of maggots as feed supplement and organic
fertilizer. The optimized maggot inoculation for the rst-stage

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F.-X. Zhu et al. / Bioresource Technology 116 (2012) 485491

Fig. 3. Temporal activity patterns of different types of microbial enzymes in pig manure composts during two-stage composting. s, Maggot-treated compost. 4, Natural
compost (control). Error bars: SD of the mean from three replicates.

F.-X. Zhu et al. / Bioresource Technology 116 (2012) 485491

Fig. 4. Germination index trends of Chinese cabbage (a) and cucumber (b) seeds in
the aqueous extracts of 10% sample suspensions during two-stage composting. s,
Maggot-treated compost. 4, Natural compost (control). Error bars: SD of the mean
from three replicates.

composting not only warranted the maggot production of at least

100 kg/ton within 1 week but shortened greatly the time length
for the second-stage composting to maturity in terms of the
changes of physical, chemical and enzymatic parameters and
phytotoxic effects monitored during the 13 week composting.
Our study highlights a high economic value of pig manure that
can be achieved through the two-stage composting without the
use of bulking agents.
Acknowledgements
The funding of this study was provided jointly by the Agricultural Ministry of China (Grant Nos.: 2011-G27 and 201103004),
the Ministry of Environmental Protection of China (Grant No.:
2008ZX07101-006), the Zhejiang Provincial R/D Program (Nos.:
2010C02001 and 2008C12045-1) and the Zhejiang Academy of
Agricultural Sciences (No. 2010CX20).
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