Environmental Technology & Innovation 25 (2022) 102193

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Environmental Technology & Innovation

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

Organic waste management by two-stage composting process

to decrease the time required for vermicomposting
∗
Heena Kauser , Meena Khwairakpam
School of Agro and Rural Technology, Indian Institute of Technology Guwahati, 781039, Assam, India

article info a b s t r a c t

Article history: A fully matured product from a traditional vermicomposting method takes 45–60 days,
Received 10 September 2021 which is 25–30 days longer than a rotary drum composting technique. Therefore, the use
Received in revised form 1 December 2021 of rotary drum compost would minimize the vermicomposting period by 15–20 days.
Accepted 6 December 2021
Very limited studies were available on combined technologies used for the biodegra-
Available online 10 December 2021
dation of invasive weeds. In this research, in-vessel composting and vermicomposting
Keywords: technologies are proven to be best in time reduction and produce mature, stable, and
Weed management nutrient-rich compost that is more superior quality than conventional rotary drum
Composting time compost. Esenia fetida, Eudrilus eugeniae, and Perionyx Ceylanesis were used to choose the
Rotary drum composting best earthworm species for surviving with a low death rate and producing high-quality
Vermicomposting
compost. For vermicomposting of pre-degraded waste, four separate reactors were set
Earthworm species
up: one with E. fetida (VrEF), one with E. euginae (VrEE), one with P. ceylanesis (VrPC), and
one sans earthworm species (VrC). Vermicompost made from waste by all earthworm
species exhibited a better nutritious value than control. Furthermore, E. fetida produced
vermicompost with total Kjeldahl nitrogen (TKN) content of 3.24%, 12.87 g/kg total
phosphorus (TP), and 22.08 g/kg potassium. The breakdown of complicated chemicals
into simpler forms is confirmed by FTIR spectra. To confirm the vermicompost product,
a germination assay was conducted on Vigna radiata. As a result, vermicomposting is the
most effective nutrient recovery method. Weed management and the usage of two-stage
compost products in the agro-industry could benefit from the current research.
© 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

Due to the increased population and rapid urbanization, solid waste management became a vital environmental threat
in the 21st century. There are viable approaches for processing of organic waste locally to convert into valuable products.
Composting is the most acceptable and viable option for organic waste management, since it is easier to operate in
enclosed areas as long as a good quality product is managed properly. Composting is the process of organic matter
degradation in the presence of oxygen under controlled conditions, resulting in a good product that can be used as a
soil conditioner for gardening, agriculture, and horticulture, among other applications (Singh et al., 2011). Composting
within vessels has demonstrated high efficiency in improving soil characteristics, like soil conductivity, stabilization,
tolerance to erosion, soil fertility, and plant nutrition (Celik et al., 2004). In-vessel composting technology is more suitable
for organic substrate degradation as it takes less space and time along with better control and high performance (Kim
et al., 2008). On the other hand, vermicomposting is also an aerobic degradation of organic compounds involving the

∗ Corresponding author.
E-mail address: heena176154002@iitg.ac.in (H. Kauser).

https://doi.org/10.1016/j.eti.2021.102193
2352-1864/© 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.
org/licenses/by-nc-nd/4.0/).
H. Kauser and M. Khwairakpam Environmental Technology & Innovation 25 (2022) 102193

use of earthworms to convert the organic compound into a humus-like substance known as vermicompost (Munroe,
2007). Composting and vermicomposting are also commonly recommended as biological waste treatment methods for
converting organic waste into usable soil conditioners (Tognetti et al., 2005). In vermicomposting, which does not require
a thermophilic process (earthworms may perish if the temperature of the overall reactor or vermicomposting system
rises over 35 ◦ C), both earthworms and microbes are closely intertwined to stabilize the organic materials (Riggle and
Holmes, 1994). Vermicomposting is a low-cost technology solution that converts agricultural waste into organic fertilizers
by allowing earthworms and microorganisms to engage throughout the worm gut (El-Haddad et al., 2014). However,
vermicomposting time ranges from 100 days (Garg et al., 2006), 90 days (Bansal and Kapoor, 2000), 75 days (Bharadwaj,
2010) respectively. The main disadvantage of vermicomposting is that it takes longer than in-vessel composting, despite
having greater consumer recognition than compost due to its higher nutritional content recovery. Before earthworm
inoculation, it is preferred to achieve thermophilic phase (pre-composting), which facilitates pathogen elimination, waste
stabilization, poor heat resistance in earthworms, the reduced earthworm mortality rate (Domínguez et al., 1997),
and a shorter vermicomposting cycle time. Therefore, composting and vermicomposting can be done in sequence. The
thermophilic phase of composting is required for mass reduction, waste stabilization, and pathogen removal before the
vermicomposting process (Frederickson et al., 2007). Many researchers have used composting and vermicomposting
technologies to solve the issue of terrestrial weeds like Parthenium hysterophorus (Rai and Suthar, 2020; Sharma and
Chowdhary, 2020; Devi and Khwairakpam, 2021), Lantana camara (Devi and Khwairakpam, 2020a,b) and (Rai et al., 2021),
Mikania micrantha Kunth (Kauser et al., 2020 and Kauser and Khwairakpam, 2021) by converting the biomass into a value-
added resource like compost or vermicompost products. Sharma and Garg (2020) described the 3-week pre-degradation
of Parthenium and buffalo dung before the vermicomposting process using E. fetida, which lasted 90 days. Therefore, most
research focused on local/regional specific invasive weeds using traditional composting techniques, but no research has
focused on a two-stage composting technique (Rotary drum composting followed by vermicomposting) for weeds like
‘mile-a-minute.’
The novelty of the current study lies in reducing the time required for vermicomposting by incorporating an in-vessel
compost technique for 10 days, followed by a 20-day vermicomposting process and obtaining nutritional ‘vermicast’
from it. The overall aim of this study is to monitor various physicochemical and biological parameters during the Rotary
followed by vermicomposting (RVC) process to compare its efficiency to that of traditional vermicomposting techniques
in terms of nutrient quality. This study employs terrestrial weed biomass ‘mile-a-minute’, also known as M. micrantha,
because no extensive research has been conducted on this weed biomass using a two-stage composting technique. It is
regarded as one of the most invasive weeds and is readily available in huge quantities in the Northeastern part of India.
Earthworms require more time for acclimatization than the actual process to happen. So a pre-degradation process is
conducted in a 550 L capacity Rotary drum composter to acquire a significant rise of temperature (thermophilic phase)
and then vermicomposting for 20 days using bamboo bins.

2. Materials and methods

2.1. Rotary drum composter

Fig. 1 shows a schematic representation of a pilot-scale rotary drum composter. The total capacity of the drum (R1)
is 550 L in which 150 kg of waste is pre-degraded for 10 days. The drum is mounted on a metal stand and rotates
mechanically with the help of four rubber wheels. The length and diameter of the drum are 1.022 m, 0.76 m, and 4 mm
metal sheets respectively. 40 mm angles are welded inside the drum longitudinally so that the waste is adequately mixed,
agitated, and aerated during rotation. Anti-corrosive material in the drum is used to prevent rusting as it might damage
the drum due to the production of leachate from waste. Two 10 cm holes adjacent to the drum are designed to drain the
surplus water.

2.2. Vermireactor design and earthworm culture

The experimental procedure was carried out in a bamboo reactor of almost 86.12 × 102 mm3 sizes. Banana leaves
that have been degraded for 1–2 weeks have been used as bedding material for vermireactor. 5:4:1 ratio is used for the
mixture of waste that has been obtained from previous study on drum composting of MM (Kauser et al., 2020). MM was
collected from the vicinity of the IIT Guwahati campus, Amingaon, Assam, India. Cow dung (CD) and sawdust (SD) were
collected from a local dairy farm nearby IITG campus and Amingaon market respectively. A total amount of 1.5 kg waste
has been used including MM as the first part, CD as the second part, and SD as the third part. For each reactor, a total of
120 earthworms were used. For the process, three separate earthworm species were used for application in the reactor
namely: applying E. fetida (VrEF), applying E. euginae (VrEE), applying P. ceylanesis (VrPC), and without earthworm species
(VrC). Wet jute bags were used to cover the vermireactor because earthworms can perform more efficiently with less light
exposure.
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Fig. 1. In-vessel composting technique (Rotary drum composter).

Table 1
Characterization of various parameters of feedstock material.
Parameters Unit MM CD SD
TOC % 48.01 ± 1.02 46.92 ± 1.4 8.19 ± 1.8
pH 5.24 ± 0.09 5.8 ± 0.1 6.8 ± 0.09
EC mS/cm 7.87 ± 0.4 3.78 ± 0.01 1.23 ± 0.1
Na g/kg 3.87 ± 0.09 2.12 ± 0.09 1.4 ± 0.2
K g/kg 5.01 ± 0.09 0.23 ± 0.1 0.8 ± 0.08
Ca g/kg 5.24 ± 0.1 8.72 ± 0.1 1.87 ± 0.09
Pb mg/kg 28.23 ± 1.1 12.12 ± 0.09 ND
Cu mg/kg 11.24 ± 0.7 17.02 ± 0.08 1.01 ± 0.8
Ni mg/kg 2.15 ± 0.2 24.23 ± 1.12 0.4 ± 0.02
Mn mg/kg 2.12 ± 0.05 10.12 ± 0.08 0.54 ± 0.01

2.3. Rotary drum followed by vermicomposting (RVC)

The waste mixture was first to feed into the Rotary drum reactor (R1) to obtain thermophilic temperature. When the
temperature drops to 35–40 ◦ C, the partially decomposed material was used for vermicomposting with three different
earthworm species, E. fetida, E. eugeniae, and P. Ceylanesis. E. fetida and E. eugeniae were obtained from Krishi Vigyan
Kendra, Kamrup Rural, Assam. However, P. ceylanesis, popularly known as ‘Jai Gopal’, is a genetically engineered species
established by IVRI, Barielly, India. The initial characterization of the feedstock material has been illustrated in Table 1.

2.4. Analyses (Physico-chemical, biological, total heavy metal content, and FTIR)

The temperature was measured in situ with a portable thermometer (Mextech Digital thermometer, Mextech, India) at
various base levels every 4 h during the first 10 days of the composting period. The sample was oven-dried (Hot air oven,
ICT, India) at 105 ◦ C for 24 h before grinding and sieved with 0.2 mm US Mesh 70 (Jain and Kalamdhad, 2018; Tiquia and
Tam, 1998). The volatile solid was measured first by igniting 5 g of the sample at 550 ◦ C (Muffle furnace, ICT, India) (EPA
196). As described by Maturi et al. (2021), total organic carbon (TOC %) was calculated using Eq. (1):

TOC (%) = VS (%) /1.83 ∗ 100 (1)

Chemical parameters like pH (µ pH system 361, 132, Systronics, India) and Electrical conductivity (EC) (Digital conduc-
tivity meter, VSI-04-Deluxe, India) was measured using aqueous extract of the sample by vortexing the same in the ratio
1:10 (w/v) for 2 h at 120 rpm (Tiquia and Tam, 1998). Nutrient content namely potassium (K), and calcium (Ca) were
measured using flame photometry (Flame photometer 128, Systronics, India). Biological parameters: sBOD was estimated
using Winkler’s method, and sCOD (COD digester, Hech DRB 200, USA) was measured using the Closed Reflux-Dichromate
method (Paul et al., 2020). CO2 evolution and oxygen uptake rate (OUR) were measured as described by Varma and
Kalamdhad (2014). Total metal content was estimated using Atomic Absorption Spectroscopy (Varian Spectra 55B, India)
by digesting (Kelplus Digestion Unit, KES 42 R, Pelican, India) 0.2 g powdered sample using di-acid (5:1 v/v of sulfuric
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acid and perchloric acid) for 2 h at 300 ◦ C (Singh and Kalamdhad, 2012). To investigate changes in the functional group,
FTIR (PerkinElmer, UK) spectra were observed using a mixture combination of 300 mg KBr and 1 mg of the sample which
is recorded in 400–4100 cm−1 wave number scans.

2.5. Germination assay

The Germination Index (GI) test was performed using a compost of reactor R1 and vermicompost (VrEF, VrEE, VrPC, and
VrC) extract, as preceded by Matthews and Hastings (1987) modified procedure. Samples were prepared by combining
100 g of a single sample with 300 mL of dH2 O and shaking mechanically in a rotary shaker for 24 h at 120 rpm to achieve
a homogenized mixture. After 24 h of continuous shaking, samples were purified, and the filtered sample was used for
phytotoxicity evaluation. With different dilutions (0%, 25%, 50%, 75% and 100% v/v), compost and vermicompost extract
solutions were formulated using dH2 O. As a test plant for phytotoxicity evaluation, V. radiata (Mung bean) was used for
analysis. Mung bean was acquired from a certified store in Assam, Guwahati. Mung bean seeds were immersed in HgCl2
solution (0.1 percent w/v) for proper sterilization for 10 min before starting the experiment and washed thoroughly with
dH2 O to eliminate traces of HgCl2 . For the experiment, glass Petri plates were used to be sufficiently moistened with the
respective All the tests were conducted in triplicates, incubated for 4–5 days at 25 ± 1◦ C. After 5 days of incubation,
seedling growth, and biomass were analyzed (8 h duration of light cycle and 16 h duration of dark cycle).

2.6. Statistical analysis

Findings were statistically analyzed in ANOVA using IBM-SPSS (v.25) at p<0.05. Every reactor was placed in triplicate,
for which mean and standard deviation was calculated. The degree of interaction between measured parameters and their
phenomenon is explained by correlation coefficient analysis. As a relationship between parameters, Pearson correlation
gives an output of −1 to +1. A positive value indicates that the measured parameters have a positive relationship, while
a negative value indicates that they have a negative relationship.

3. Results and discussion

3.1. Evolution in temperature

Microorganisms play an important role in composting, whereas earthworms play a major role in vermicomposting.
Vermicomposting takes place at room temperature instead of composting, which has a thermophilic phase that ensures
material sanitation and maximum degradation (Fornes et al., 2012). In the present experimental setup, a temperature
profile was recorded to evaluate the completion of the thermophilic stage. The maximum temperature reached during
this stage was 54.3 ◦ C on day 2 in the reactor R1 as shown in Fig. 2. The thermophilic stage was almost over by day 6.
But the rotary drum composting was continued till day 10 to ensure further degradation of the organic compounds and
to obtain the optimum temperature required for the earthworms. The temperature begins to drop after the fourth day
of composting, and on the tenth day, it was measured at 37–38 ◦ C, which is advisable for the vermicomposting process
(Reinecke and Kriel, 1981). The rate of oxidative decomposition of various waste is greatest during composting in the
thermophilic phase of temperature range 50–60 ◦ C (Varma and Kalamdhad, 2015) which may reduce the time required
for vermicomposting as maximum reduction occurs during this phase and earthworm gets pre-degraded material, which
is more useful for making better quality vermicompost. Similar research was conducted on a combination of both the
composting technologies for the stabilization of cattle manure (Vicentin et al., 2021).

3.2. Evolution in pH and EC

The substrate used in the vermicomposting process has a significant impact on pH variation. Deviation of pH towards
acidic and alkaline conditions during the process is responsible for the production of acids and NH3 based on the raw
material used (Varma et al., 2015). In reactor R1, pH rose from 7.20 to 7.90 during the first 10 days. The final pH value
was in the range of 7.60, 7.99, 7.94, and 7.97 for Vr EF, Vr EE, Vr PC, and Vr C respectively (Fig. 3a). The pH values during
the vermicomposting had a decreasing trend. Sharma and Garg (2020) achieved a comparable neutral pH value while
vermicomposting Parthenium plants after 90 days of vermicomposting (pre-degradation for 3 weeks). The author reported
−,
that pH between 7–7.5 is mostly obtained due to the formation of PO34− , NO3 and CO2 emissions. Sharma et al. (2021) also
reported that the condition is mainly gained due to various compounds secreted by earthworms, that play a significant
role in neutralizing the R-CO2 H group and C6 H5 OH group. pH in all the reactors was in the range of acceptable range of
6.5–8.0 (Zhang and Sun, 2015). In addition to pH, EC is an important parameter when compost toxicity is seen, as EC above
4ds/m for application of soil is not recommended (Karak et al., 2013). The initial EC values were found to be 5.42 mS/cm,
which was reduced to 5.08 mS/cm in the reactor R1 whereas during vermicomposting the final values were reduced to
3.21, 3.78, 3.45, and 4.65 mS/cm in the reactor Vr EF, Vr EE, Vr PC, Vr C as shown in Fig. 3b. The EC values of all the samples
after 30 days were lower than the initial days, which could be due to organic acid decomposition leading to higher pH
and precipitation of soluble salts. At p<0.05, pH and EC differed significantly between tests during the RVC process.
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Fig. 2. Temperature profile during rotary drum composting.

Fig. 3. Transformation in (a) pH and (b) EC during RVC process.

3.3. Evolution in TOC and CO2 evolution

Varying physico-chemical changes attributed to microbial and earthworm actions throughout the vermicomposting
process result in a reduction of volatile solids as well as total organic carbon (TOC). Microorganisms usually utilize
substrate for the building of cells (anabolic reaction) and the production of energy (catabolic reaction). Throughout the
vermicomposting process reduction of TOC was observed due to the degradation of organic matter by earthworms and
microorganisms in all the reactors as illustrated in Fig. 4a. Earthworms contain a variety of enzymes such as proteases,
lipases, cellulases, amylases, alkaline phosphatases, dehydrogenases, and ureases that contribute to the breakdowns of
organic compounds (Usmani et al., 2019). In reactor R1 initial TOC was observed to be 37.6% that has further declined to
32.27% at the end of 10 days having a thermophilic phase of almost 4 days. Reduction of TOC for the reactor VrEF, VrEE,
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Fig. 4. Transformation in (a) TOC and (b) CO2 evolution during RVC process.

VrPC, and VrC was recorded as 28.69, 27.32, 28.89, and 29.81% respectively during the first 10 days of vermicomposting.
At the end of the vermicomposting process, TOC reduction was observed as 19.50, 21.73, 22.48, 27.85% for the reactor
VrEF, VrEE, VrPC, and VrC respectively. (Devi and Khwairakpam, 2020a,b) reported a 38.5% reduction in TOC after 45
days of vermicomposting with the invasive weed A. conyzoides. In the current study, a remarkably similar result was
observed within 30 days of the period. Organic material degradation produces CO2 , and CO2 emissions represent a reliable
microbial indicator of composting activity (Zhong et al., 2020). During the first 10 days of rotary drum composting, the
rate of respiration reduced by 40.69%. The reduction during vermicomposting appeared to be consistent across all trials;
conversely, as evidenced by the results, various substrate combos tend to influence the earthworm species. CO2 evolution
in the vermireactors was observed to be decreasing, as shown in Fig. 4b. The breakdown of organic matter is primarily
represented by the amount of O2 consumed and CO2 released, which is dependent on the chemical composition of the
substrate used. CO2 evolution was observed to be decreasing as the TOC decreased during the RVC process. van Ginkel
(1996) described the catabolic and anabolic reaction that takes place during the degradation of the organic compound.
The reaction is shown as follows where Eq. (2) represents a catabolic reaction, (3) represents an anabolic reaction, and
Eq. (4) represents the combination of catabolic and anabolic reaction.

(1 − A) Csubstrate + (1 − A) O2 → (1 − A) CO2 + (1 − A) e1 (2)

A Csubstrate + A e2 → A Cmicro (3)
Csubstrate + (1 − A) O2 → AC micro + (1 − A) CO2 + {(1 − A)e1 − Ae2 } (4)
Where,
e1 denotes the catabolic reaction’s energy in kJ mol C−1
e2 denotes energy required to build microbial bodies from organic biomass in kJ mol C−1
and, A represents the yield in microbial-C/ mol of substrate-C.
On the right-hand side of the equation in reaction (4), the term (A Cmicro ) describes the microbial cell yield, the second
term ((1-A) CO2 ) describes the CO2 yield, and the third term {(1-A) e1 – A. e2 } describes the heat produced during the
degradation. In this study, during the first 10 days of composting process microbes plays an important role where the
maximum amount of heat is produced due to the degradation of the organic compound, after 10 days earthworms play
an important role to make the mixture stable and mature. The RVC process has helped to achieve that within 30 days
because the conventional vermicomposting process takes more time to degrade organic compounds. At p<0.05, TOC and
CO2 evolution differed significantly between tests during the RVC process.

3.4. Evolution in TKN, total phosphorus content (TP), and nutrient content

Compost quality is determined in part by the amount of nutrients present. Reactor Vr EF, Vr EE, Vr PC had significantly
higher nutrient contents (p<0.05). TKN content was increasing throughout the process of vermicomposting process. During
the first 10 days of rotary drum composting slight increase of TKN was observed that may be due to the reduction
of substrate. After 10 days of rotary drum compost, the partially degraded substrate was used for vermicomposting
for almost 20 days. At the end of the first 10 days of vermicomposting, TKN was recorded as 2.54, 2.03, 1.91, 1.59%
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for the reactor Vr EF, Vr EE, Vr PC, and Vr C respectively as shown in Fig. 5a. It was seen that after the next 10 days of
vermicomposting it increased to 3.24, 2.89, 3.02, and 1.79%. The current study attained 3.24% (reactor VrEF) of TKN
within 30 days of vermicomposting process. Total nitrogen increased during the first ten days which was due to a
decrease in the carbon content of the substrate as a result of CO2 loss, as well as the action of azotobacteria that fix
nitrogen from the atmosphere (Zorpas et al., 2003). An increase of TKN was also observed due to earthworm activity
that inclines the nitrogen production through microbially mediated nitrogen transformation by addition of mucus and
nitrogenous substances by earthworm (Gunadi et al., 2002). The author further reported that nitrogen is abundant in
amino acids, which are the basic components of proteins and nucleic acids. Ammonia, which can be procured from the
breakdown of protein in the compost, is used to meet the microbial need for nitrogen that is mostly done by hydrolyzing
enzyme protease. Nitrogen is then retrieved from the carbon chain as NH+ 4 -N, which is volatilized and released into the
atmosphere (van Ginkel, 1996). MM, cow dung, and sawdust is good source of protein that was degraded by help pf
earthworms and microorganisms during the process. Devi and Khwairakpam (2020) observed a similar trend of increase
of TKN during vermicomposting of Ageratum conyzoids using E. fetida. Increment in TKN may be also due to the loss
of organic compounds as CO2 as well as water loss through evaporation (Viel et al., 1987). All the earthworm species
have shown positive results on the degradation of organic compounds and increment of TKN throughout the process of
vermicompost. Total nitrogen is typically influenced by proteolytic bacteria as well as the temperature where nitrogen is
released into the atmosphere through volatilization (Wagner et al., 1990). Similarly, mineralization of organic phosphorus
increases TP during composting process Fig. 5b. An increase in the concentration of TP was observed in all the trials
throughout the process. Earthworms pass organic materials through their guts, which transform phosphorus into a form
that can be used by plants (Lee, 1992), and earthworms are benefited from the materials that are pre-degraded as they can
easily digest them fast. The pre-degraded waste material has helped earthworms for proper digestion and acclimatization
eventually increasing the nutrient content. Similar findings have been reported, with the augmentation of TP and its
mineralization during vermistabilization of organic substrates is being attributed to phosphatase and phytase activity by a
gut microorganism of earthworm (Parthasarathi et al., 2016). Maximum TP was recorded for the reactor VrEF (12.87 g/kg)
followed by VrEE (10.45 g/kg) and VrPC (9.45 g/kg). It was observed that in the reactor VrC a gradual decrease of TP
was recorded which may be due to leaching of the nutrient from the vermireactor. Since the substrate was pre-degraded
before the vermicomposting process, cutting the vermicomposting time did not affect the final product’s nutrient content.
The concentration of Na increased from 1.53 to 2.02 g/kg during the rotary drum composting. The final concentration
of Na after vermicomposting was 2.81, 2.34, 2.58, and 2.41 g/kg for Vr EF, Vr EE, Vr PC, and Vr C respectively at the end of
the 30th day (Fig. 5c). K concentration was also found to be increasing throughout the process. K concentration increased
from 19.89 to 20.72 g/kg during 10 days of rotary drum composting. During the vermicomposting phase, the value of
potassium concentration increased to 26.78, 22.28, 24.65, and 22.14 g/kg in Vr EF, Vr EE, Vr PC, and Vr C respectively at the
end of 30 (Fig. 5d). The value of Ca concentration increased from 1.54 to 1.82 g/kg during the first 10 days of rotary
drum composting. The concentration of Ca after the vermicomposting was found to be 3.16, 3.86, 3.45, and 2.45 g/kg
for Vr EF, Vr EE, Vr PC, and Vr C respectively at the end of 30 (Fig. 5e). The action of pH change and microbial activity on
organic matter as it passes through the earthworm gut can be attributed to an increase in macronutrients (Kouba et al.,
2018). Varma et al. (2016) also reported the rise of nutrient content during vermicomposting of the aquatic weed water
hyacinth. In terms of nutrient concentration, reactor VrEF has demonstrated better results than other reactors. p<0.05,
nutrient content differed significantly between tests during the RVC process.

3.5. Evolution in sBOD, sCOD, and Oxygen uptake rate (OUR)

The decreasing trend in the concentration of sBOD was observed in all the trials during the RVC process that
is an important parameter for compost quality (Kalamdhad and Kazmi, 2007). For reactor R1, it was observed that
sBOD decreased to 208.42 mg/L from 356.5 mg/L after the first 10 days. However, when the pre-degraded waste was
vermicomposted, it was observed that the sBOD concentration decreased from 208.42 mg/L to 46.4 mg/L for reactor Vr EF,
208.42 mg/L to 90.2 mg/L for reactor Vr EE and 208.42 mg/L to 72.2 mg/L for reactor Vr PC as shown in Fig. S1(a). sBOD
reduction during the RVC process was due to the degradation of an organic compound by earthworm and microorganisms
that attends stabilization which can be further used for the crop in soil (Wang et al. 2004). Comparisons among the
different techniques and trials with rotary drum composting and vermicomposting showed that sCOD was decreased from
672 mg/L to 576 mg/L in the first 10 days for reactor R1. It was reduced further to 117.34 mg/L, 181.34 mg/L, 202 mg/L
and 240 mg/L for the reactor Vr EF, Vr EE, Vr PC and Vr C respectively as shown in Fig. S1(b). As described by Jain et al. (2018),
organic matter deterioration determines whether sBOD and sCOD increase or decrease. Biological content reduction
reduces sBOD and sCOD, reducing emissions of carbon dioxide, suggesting the compost has undergone stabilization. sBOD
and sCOD analyzed varied differently at p<0.05 during the RVC process. OUR was reduced by 44.87% in reactor R1 during
the first 10 days, and 90.96% in VrEF, 89.54% in VrEE, 87.57% in VrPC, and 69.39% in VrC during the RVC process. OUR is the
most appropriate parameter for determining compost or vermicompost stability. Microorganisms will consume organic
matter for their metabolic process which results in high oxygen consumption rates conferring compost instability (Said-
Pullicino et al., 2007). As the organic matter decreases, less consumption of oxygen will occur by a microorganism that
confers compost stability. The rate of OUR evolution was minimum in the first 10 days of composting process, however,
it was increased once the pre-degraded waste was vermicomposted where earthworm and microorganisms helped in the
degradation of organic matter resulting in a decrease of OUR (Paul et al. 2020). Since conventional vermicomposting takes
more than 45 to 60 days, the current study not only reduced the duration but also achieved OUR value >1 for the VrEF
reactor followed by VrEE and VrPC, which shows vermicompost stability within 20 days as shown in Fig. S1(c).
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H. Kauser and M. Khwairakpam Environmental Technology & Innovation 25 (2022) 102193

Fig. 5. Transformation in (a) TKN and (b) Total phosphorus (c) Sodium (d) Potassium and (e) Calcium during RVC process.

3.6. Total heavy metals (HM)

In all the reactors, four different HMs (Pb, Cu, Ni, and Mn) were examined. In reactor R1, Pb was found to be 15.75
mg/kg which was increased to 24.17 mg/kg after 10 days. After the vermicomposting of the pre-degraded waste, Pb was
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H. Kauser and M. Khwairakpam Environmental Technology & Innovation 25 (2022) 102193

Table 2
Concentration of heavy metals during RVC process.
Lead (Pb) (mg/kg)
Days R1 Vr C Vr EE Vr EF Vr PC
0 15.75 ± 0.09 – – – –
10 24.17 ± 0.007 – – – –
20 – 25.74 ± 0.02 25.89 ± 0.02 24.75 ± 0.09 26.78 ± 0.04
30 – 26.78 ± 0.05 27.34 ± 0.04 26.14 ± 0.07 27.74 ± 0.02
Copper (Cu) (mg/kg)
0 49.87 ± 0.07 – – – –
10 50.10 ± 0.05 – – – –
20 51.42 ± 0.08 52.10 ± 0.005 48.98 ± 0.07 51.25 ± 0.02
30 51.89 ± 0.02 54.78 ± 0.04 49.52 ± 0.09 52.47 ± 0.04
Nickel (Ni) (mg/kg)
0 24.02 ± 0.07 – – – –
10 27.45 ± 0.1 – – – –
20 – 30.23 ± 0.06 31.54 ± 0.05 30.98 ± 0.07 30.21 ± 0.07
30 – 34.23 ± 0.07 32.21 ± 0.06 34.56 ± 0.02 36.43 ± 0.05
Manganese (Mn) (mg/kg)
0 44.85 ± 0.07 – – – –
10 47.4 ± 0.06 – – – –
20 – 55.87 ± 0.02 61.7 ± 0.08 53.43 ± 0.1 52.72 ± 0.04
30 – 57.42 ± 0.04 62.5 ± 0.04 58.78 ± 0.02 54.87 ± 0.05

increased at the end of the 30th day for all reactors. Cu, Ni, and Mn were also found to increase in all the reactors after
the end of the RVC process. As discussed by Yadav and Garg (2011), the increase of HM in the final compost is due to the
release of CO2 during the process that lowers down the total weight of the compost and increases its material content.
Table 2 describes the total concentration of HM on Day 0, day 10, day 20, and day 30 of the RVC process for all the
reactors. The increase in HM in all the reactors as illustrated in Table 3 for the reactor R1, Vr EE, Vr EF, Vr PC, Vr C was due
to the loss of organic content that attributed to the reduction of total weight and increase of material contained in the
final compost. Balanchandar et al. (2021) observed similar results for the vermicompost of Ipomoea staphylina, an invasive
weed. The dynamic increase in HM, according to the authors, is due to the loss of TOC and weight reduction by degradative
microorganisms and earthworms. The increased change of HM during vermicomposting of Parthenium hysterophorus has
been well documented (Yadav and Garg, 2011) which is in support of the present study. The concentration of all HMs
was within the permissible limit of the Fertilizer Control Order (FCO) (NEERI, 2009). At p<0.05, heavy metals differed
significantly between tests during the RVC process.

3.7. Germination test (Vigna radiata)

Germination index (GI) is an important parameter to assess the phytotoxicity of vermicompost. It is always recom-
mended that a GI value exceeding 80% indicates a very good and healthy compost product (Wang et al., 2021). An
exploration of the phytotoxicity of various concentrations of different phased vermicompost samples was performed using
mung bean (V. radiata) germination test for 5 days. GI (%) for the reactors inoculated with earthworms were in the range
of 80%–100% for all the concentrations of vermi extract whereas GI% for reactor Vr C was in the range of 50%–90% where
after 50% concentration the growth of mung bean gradually decreased as illustrated in Table 3. A combined review of
physicochemical analysis and toxicity tests is more useful for estimating the consistency of compost. In reactor R1, the
exclusion of root and shoot growth of V. radiata indicates the presence of toxic compounds. Given that there is sufficient
evidence that MM contains some allelopathic compound that has a negative effect (Ullah et al., 2014), the decrease in GI%
as the concentration of extract increased in the untreated waste mixture could be attributed to the allopathic compounds
that it contains. The authors also reported a similar study in which the extract of MM inhibited the growth of Fymbristylis
population. Li and Jin (2010) reported about water-soluble phytochemicals present in MM that inhibit the germination of
Coix lacryma-jobi. There was a significant increase in root length and shoot length for the reactor Vr EE, Vr EF, Vr PC as the
pre-degraded waste was vermicomposted where the degradation was mostly done by earthworms and its gut bacteria. The
length was highest in the reactor Vr EF as compared to the other two reactors inoculated with earthworms. The increasing
growth of root length and shoot length signifies the reduction of toxicity during the vermicomposting process. Table 4
shows the GI %, root length, and shoot length of V. radiata at different aqueous extract concentrations. The germination
test differed significantly among the reactors at p<0.05.

3.8. Growth and development of earthworm

The development of earthworm species used in the vermiconversion of organic waste materials is a good indicator
of a successful vermicomposting process. The current study emphasized the growth and breeding of earthworm species
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H. Kauser and M. Khwairakpam Environmental Technology & Innovation 25 (2022) 102193

Table 3
Germination Index test for compost produced from RVC process.
Rotary drum compost R1 (Day 0)
Sample (%) Germination percentage (%) Root length (cm) Shoot length (cm)
0 100 ± 0 5.1 ± 0.02 2.4 ± 0.08
25 73.3 ± 1 1.3 ± 0.08 1.8 ± 0.09
50 63.3 ± 0.5 1.4 ± 0.07 1.9 ± 0.07
75 36.6 ± 0.8 1.2 ± 0.1 0.5 ± 0.05
100 26.6 ± 0.9 1.2 ± 0.02 0.7 ± 0.05
Vr C (Day 30)
0 100 ± 0 5.1 ± 0.1 2.2 ± 0.1
25 76.6 ± 0.05 1.8 ± 0.04 2.1 ± 0.09
50 70 ± 0.02 1.1 ± 0.02 1.7 ± 0.07
75 56.6 ± 0.07 1.2 ± 0.04 1.5 ± 0.08
100 43.3 ± 0.02 1.0 ± 0.05 1.2 ± 0.03
Vr EE (Day 30)
0 100 ± 0 5.1 ± 0.07 2.4 ± 0.07
25 93.3 ± 0.1 3.9 ± 0.04 3.5 ± 0.03
50 96.7 ± 0.08 4.5 ± 0.08 2.9 ± 0.08
75 86.6 ± 0.04 5.7 ± 0.09 3.2 ± 0.1
100 87.6 ± 0.02 4.8 ± 0.1 3.1 ± 0.2
Vr EF (Day 30)
0 100 ± 0 5.1 ± 0.09 2.4 ± 0.1
25 100 ± 0.02 3.6±.0.08 3.2 ± 0.08
50 96.6 ± 0.2 4.2 ± 0.2 4.2 ± 0.04
75 93.3 ± 0.04 5.5 ± 0.04 3.6 ± 0.05
100 96.7 ± 0.08 4.8 ± 0.06 3.2 ± 0.2
Vr PC (Day 30)
0 100 ± 0 5.1 ± 0.07 2.4 ± 0.05
25 96.7 ± 0.04 3.4 ± 0.05 2.9 ± 0.02
50 80 ± 0.08 4.5 ± 0.1 3.1 ± 0.08
75 73.3 ± 0.02 5.2 ± 0.02 2.9 ± 0.07
100 73.3 ± 0.04 4.6 ± 0.04 1.8 ± 0.02

E. fetida, E. euginae, and P. Ceylanesis. Except for the reactor VrPC, where there was a decline in earthworm growth, the
other two species showed an increasing trend in the growth and development during vermicomposting. A typical vermi-
composting process takes 45–60 days for full maturity and the growth of earthworm species varies accordingly. However,
in the current study due to pre-degradation of waste, earthworms could feed it easily without much acclimatization
for which the number of worms increased more rapidly within 30 days. At the end of the 30th day, earthworms were
evaluated in terms of growth rate, cocoon production, juvenile and adult number (Table 4). It has been observed that in the
reactor VrEF, the number of adult earthworms increased to 341 (juvenile 116 and cocoons 23/100 g) at the end of 30 days
followed by reactor VrEE with 296 (juvenile 96 and cocoons 14/100 g) number of earthworms at the end of 30 days. In
reactor VrPC, earthworm number was decreased to 112 (juvenile 18 and cocoons 11/100 g) at the end of 30 days and it was
seen that despite feeding degraded material, earthworms were seen coming out of the reactor during the initial days of
the process. The mortality of P. Ceylanesis could be due to the weed biomass that has been mixed with CD and SD, making
it unsuitable for it to survive. An upsurge in the metabolic activity of microorganisms associated with the earthworm’s
gut, as well as an escalation in the growth of earthworm biomass, characterize the mesophilic hydrolytic stage of the
vermicomposting process (Vivas et al., 2009). Throughout the vermicomposting process, earthworm species increased in
number, with E. fetida showing a large number of adults, cocoons, and juveniles. Pre-degrading or thermo-composting of
biomass helped in better nourishing and growth of the earthworms.

3.9. Correlation coefficients of different parameters during the composting process

Correlation is the statistical technique that indicates the linear association of variables. The coefficient varies from
+1 (high positive) to −1 (high negative) (Reddy et al. 2020). Correlation coefficients of different parameters during the
composting process for the reactor Vr EE, Vr EF, Vr PC, Vr C has been briefly represented in Table S1. For the reactor Vr EF, it
was observed that there is a strong correlation between TOC and CO2 evolution (−0.98), because of the degradation of
organic matter, carbon is released in the form of CO2 that gets stable once the organic matter is degraded and there is a
lesser amount of organic matter for the microbes. A similar trend was also followed in other reactors during the process.
Nutrient parameters like TP (−0.98), K (−0.95), Na (−0.99), Ca (−0.96), and TKN (−0.89) had a strong correlation with
TOC, as TOC is decreased all these parameters have seen to be increased. A similar trend has also been observed with other
reactors except for Vr C that showed a moderate correlation with TOC, which may be due to low degradation of organic
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H. Kauser and M. Khwairakpam Environmental Technology & Innovation 25 (2022) 102193

Table 4
Growth and development of earthworm species.
Eisenia fetida Adults Juveniles Cocoons (Nos. per
100 grams)
Day Vr EF
10 120 0 0
20 198 ± 5.56 33 ± 4.54 9 ± 1.95
30 341 ± 2.45 116 ± 2.53 23.67 ± 5.31
Eudrilus eugeniae Adults Juveniles Cocoons (Nos. per
100 grams)
Days
10 120 0 0
20 170 ± 2.62 22 ± 2.45 24 ± 2.41
30 296 ± 7.21 96 ± 3.68 14 ± 1.92
Perionyx ceylanensis Adults Juveniles Cocoons (Nos. per
100 grams)
Days
10 120 0 0
20 138 ± 4.21 10 ± 0.94 12 ± 2.05
30 112 ± 2.05 18 ± 2.16 11 ± 2.49

matter. pH has a strong negative correlation with sBOD and sCOD, as in when sBOD and sCOD decrease pH increases in
all the reactors.

3.10. Fourier transform infrared spectroscopy (FTIR)

The FT-IR spectra of vermicompost using earthworm species and without earthworm has been shown in Fig. S2. O-
H stretching was observed at 3685 cm−1 and 3306 cm−1 that signifies a strong bond that represents mainly phenolic
compounds (Hussain et al., 2015). The next peak at 2992 cm−1 is attributed to aliphatic C–H stretching of fatty acids and
lipids (Hussain et al., 2016). The peak at 1490 cm−1 indicates the presence of C==C aromatic compounds in the sample
(Mochochoko et al., 2013). The peak at 1034 and 842 cm−1 , according to the author, primarily represents C–O stretch
and CH in-plane deformation, which is consistent with our findings. The peaks in the reactor VrEF are shallower than in
the other reactors after treatment, owing to degradation by various types of enzymes found in the earthworm gut and
microflora The FT-IR graphs show a significant reduction during the vermicomposting process.

3.11. Practical implication of the study

In North-eastern India, MM’s spread has mostly impacted rubber, tea, and banana plantations, and locals have not
established a control strategy to remove this invasive weed. This weed can be managed on-site without discarding into
low-lying areas. So, local people could use the biomass for the production of compost and utilize in agricultural fields to
increase the crop yield of a variety of plants, including vegetables and fruits to attain a sustainable livelihood. There is a
paucity of research on MM composting and vermicomposting, as well as field studies of the compost, which could aid in
the paradigm change in the management of such an invasive terrestrial species. This biomass compost can be a substitute
for chemical fertilizer in the application of agricultural fields and has the potential to be used for the soil remediation
process as well.

4. Conclusion

The current study examined the use of rotary drum composting for pre-degrading waste to achieve a thermophilic
temperature, followed by 20 days of vermicomposting to improve the compost quality and shorten the vermicomposting
time. Through the two-stage composting process, E. fetida achieved 3.24% TKN with considerable decreases in organic
carbon and enhanced total phosphorus up to 12.87 g/kg. The reduction in sBOD, sCOD, and CO2 evolution rate was found
to be greater in the earthworm-aided reactor. The earthworm growth and development were higher for the species E.
fetida and E. eudrilus. After exposure to V. radiata vermicompost extract, seed germination increased, indicating that the
phytotoxic chemicals in the weed biomass may have transformed into less toxic compounds. The FTIR spectra showed a
significant transition of the complex organic compound into a simpler form. Heavy metal was significantly immobilized
with the degradation of organic compounds during the process of vermicomposting. The two-stage composting technology
could be an additional strategy to the conventional composting process in terms of quality improvement and time
reduction. The further recommendation would be an application of this weed compost for pot and field study using
different plant models in terms of plant growth and fruit quality.
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H. Kauser and M. Khwairakpam Environmental Technology & Innovation 25 (2022) 102193

CRediT authorship contribution statement

Heena Kauser: Data curation, Conceptualization, Investigation, Writing – original draft, Methodology, Software,
Validation. Meena Khwairakpam: Conceptualization, Supervision, Writing – review & editing, Visualization, Project
administration.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have
appeared to influence the work reported in this paper.

Acknowledgments

The authors are thankful to SART, IIT Guwahati, Assam, India, for providing the space and helping in the smooth
running of research work. The authors acknowledge Mr. Chejarla Venkatesh Reddy, Department of Civil Engineering,
Indian Institute of Technology, for helping throughout this research work. The author also acknowledges the DST-SERB,
Government of India for the financial help (File No. -EEQ/2018/000357). The authors are thankful to Central Instrument
Facility, IIT Guwahati for providing a platform for FTIR analysis.

Appendix A. Supplementary data

Supplementary material related to this article can be found online at https://doi.org/10.1016/j.eti.2021.102193.

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