Journal of Environmental Management 293 (2021) 112817

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Journal of Environmental Management

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

Degradation and accumulation rates of fresh human excreta during

vermicomposting by Eisenia fetida and Eudrilus eugeniae
Michael Nyame Acquah a, Helen Michelle Korkor Essandoh a, *, Sampson Oduro-Kwarteng a,
Eugene Appiah-Effah a, Peter Antwi Owusu b
a
Regional Water and Environmental Sanitation Centre, Kumasi, Department of Civil Engineering, Kwame Nkrumah University of Science and Technology, Kumasi,
Ghana
b
Biological Filters and Composters Limited (Biofilcom), Ghana

A R T I C L E I N F O A B S T R A C T

Keywords: This study was carried out to assess the degradation and accumulation rates of fresh human excreta and how
Human excreta degradation addition of anal cleansing materials affect performance during vermicomposting by Eisenia fetida and Eudrilus
Vermicomposting eugeniae. Vermicomposting setups consisting of two transparent containers (length 0.27 m, breadth 0.17 m and
Eisenia fetida
depth 0.12 m) stacked on top of each other were installed and operated under laboratory conditions. Earth­
Eudrilus eugeniae
worms, Eisenia fetida and Eudrilus eugeniae, were obtained from Green Cycle Technologies Limited in Accra,
Anal cleansing material
Helminth eggs Ghana and the species verified and cultured before use. Fresh human excreta for feeding the experimental setups
was collected from an Enviro-loo public toilet and 13 g applied to the setups daily. Similar setups were fed with
0.3 g of anal cleansing material in addition to the fresh excreta. A setup without any earthworms (NW) was set up
as a control. Physicochemical characteristics of vermicompost accumulating in the setups were determined
weekly for 28 days using standard laboratory procedures while the sludge accumulated in the various setups
were weighed and recorded daily. One-way analysis of variance (ANOVA) and Post-hoc LSD tests were conducted
to determine whether the differences in results between the two earthworm species and among setups with and
without anal cleansing materials were statistically significant (p ≤ 0.05). The study showed 12.3% and 26.2%
reduction in volatile solids in EE (Eudrilus eugeniae and excreta only) and EF (Eisenia fetida and excreta only)
while ash content increased indicating good degradation. The percentage mass reductions recorded at the end of
the fourth week were 67.5%, 58.8% and 40.5% in systems EE, EF and NW respectively, while reductions of
73.7% and 68.5% were realized in EEA (Eudrilus eugeniae with excreta and toilet paper) and EFA (Eisenia fetida
with excreta and toilet paper) respectively. There was greater amount of sludge accumulated in systems without
earthworms, 0.00020 m3 (59.5%) in NW, than in systems with earthworms where 0.00011 m3 (32.5%) and
0.00014 m3 (41.2%) were recorded for EE and EF respectively. The rate of accumulation in vermibeds EE and EF
was relatively higher than in vermibeds with excreta and toilet paper (EEA and EFA). EEA and EFA recorded
accumulation rates of 0.00009 m3 (26.3%) and 0.00011 m3 (31.5%) respectively. Complete removal of helminth
eggs was not achieved in any of the treatment systems.

1. Introduction 2017). In addition to the high dependency on OSS facilities, high

Most densely populated urban regions without adequate sanitation
and sewerage infrastructure face major problems in the management of
human excreta (Strauss et al., 2000). Human excreta if not managed
properly can cause waterborne diseases and other health implications. It
is estimated that about 2.8 billion people in low and middle income
areas are served by onsite sanitation systems (OSS) (WHO and UNICEF,
financial cost involved in transportation of faecal sludge to designated
disposal sites, due to the distance and traffic congestion in most urban
cities, often leads to uncontrolled dumping of faecal sludge at unap­
proved sites (Ingallinella et al., 2002).
The problem with onsite sanitation is the incomplete onsite treat­
ment of the excreta for safe removal and disposal. Thousands of tons of
untreated sludge from various OSS installations are indiscriminately

* Corresponding author. Department of Civil Engineering, Kwame Nkrumah University of Science and Technology Kumasi, Ghana.
E-mail address: hmkessandoh.coe@knust.edu.gh (H.M.K. Essandoh).

https://doi.org/10.1016/j.jenvman.2021.112817
Received 9 September 2020; Received in revised form 16 May 2021; Accepted 16 May 2021
Available online 4 June 2021
0301-4797/© 2021 Published by Elsevier Ltd.
M.N. Acquah et al.
disposed of into drainage systems, open spaces and wetlands (Appia­
h-Effah et al., 2014). Helminth eggs found in faecal sludge are trans­
mitted by direct contact with contaminated soils, crops or wastewater
and they usually cause asymptomatic chronic infections (Salazar-Cas­
tanon et al., 2014). Heavy infection is associated with intestinal damage
with loss of nutrients resulting in impaired mental and physical growth,
anaemia, low resistance to other infections in children and poor birth
outcomes of pregnant women (Hotez, 2008). Faecal sludge needs to
undergo some form of treatment to reduce the pH, pathogen level,
chemical oxygen demand and biochemical oxygen demand before dis­
charging into water bodies to prevent oxygen depletion in the water
bodies. Oxygen depletion occurs as a result of degradation of organic
materials in the waste by microorganisms (Bhise and Anoakor, 2015)
and has detrimental effects on the survival of aquatic organisms. In
several aspects of waste management, regeneration of valuable re­
sources is now acknowledged (Suthar, 2010). Use of biological proced­
ures such as waste stabilization ponds, unplanted wetland drying beds
and composting/co-composting are among the effective and less
expensive ways of treating waste (Koné et al., 2010).
Dry sanitation facilities such as composting and ventilated improved
pit latrines are appropriate for communities with basic water supply, but
there are problems with odour nuisance and rapid filling of pits due to
higher usage rate. The emergence of vermi-technology in treating
human waste will help reduce the numerous effects of poor sanitation.
Vermicomposting is a method of decomposition of excreta involving the
shared action of earthworms and microorganisms. Biochemical break­
down of organic matter is often attributed to the processes and action of
microorganisms but earthworms fuel the process by conditioning and
fragmenting the substrate hence changing its biological activity (Dom­
inguez and Edwards, 2004). Despite so many earthworm families, not all
species are appropriate for vermicomposting. Some particular features
of biology and ecology make an earthworm appropriate. It should be
able to colonize organic waste naturally, have a high rate of organic
matter digestion and assimilation, and tolerate a broad variety of envi­
ronmental variables. Few earthworm species have the above charac­
teristics (Medany and Yahia, 2011). These include epigeic species such
as Eudrilus eugeniae, Eisenia fetida and Perionyx excavates. For successful
vermicomposting, suitable earthworm species selection is a vital step
which cannot be overlooked, especially its potential to feed upon
different streams of wastes. The vermicomposting efficiency of a few
epigeic earthworm species such as Eisenia fetida and Lumbricus rubellus in
the digestion of a broad range of organic materials such as animal
excreta, sewage sludge, human faeces, residual crops and agricultural
waste, raw organic waste in small and large projects has been studied
(Garg et al., 2006; Yadav et al., 2010). However, the potential of some
commonly known earthworms such as Eudrilus eugeniae for the degra­
dation of human excreta together with anal cleansing material has not
been extensively explored.
While focusing on improving sanitation technologies, less priority is
often given to the management of anal cleansing material used after
visiting the toilet. There is however a need to manage anal cleansing
material to prevent the spread of diseases (McMahon et al., 2011). Anal
cleansing materials are often not disposed of with excreta but contained
separately and burnt or added to municipal solid waste, which ends up
in landfills or the open environment. Anal cleansing materials are
managed separately due to the use of unconventional anal cleansing
materials, user perceptions over excreta containment structures filling
up quickly or upon advice from some installers of vermicomposting
toilets. So far, extensive studies have not been carried out on vermi­
composting of human excreta. Neither has the effect of anal cleansing
materials on the process been documented. This study was therefore
undertaken to assess the degradation and accumulation rates of human
excreta and the effect of anal cleansing materials during vermi­
composting by Eisenia fetida and Eudrilus eugeniae.
2
Journal of Environmental Management 293 (2021) 112817
2. Study area
Human excreta was collected from a public toilet (an Enviro-loo)
located in Kotei, a community within the Oforikrom Municipal Assem­
bly of the Ashanti Region of Ghana (Figure S1). The climate condition of
Kotei is tropical and humid with an average temperature of about 25 ◦ C.
Kotei is inhabited by both indigenes and students from the Kwame
Nkrumah University of Science and Technology (KNUST). The com­
munity is unplanned with low sanitation coverage with the populace
mostly being petty traders with few having white collar jobs. A large
proportion, about 47%, of Kotei’s residents but up to 80% in the older
residential areas do not have access to sanitation facilities in their homes
and depend on the four public toilets in the community. These facilities
are old, poorly maintained and a public health nuisance, with residents
on average walking 600 m to use them (Leathes, 2012). Vermi­
composting setups were installed and operated under laboratory con­
ditions at the Environmental Quality Engineering (EQE) laboratory of
the College of Engineering, Kwame Nkrumah University of Science and
Technology (KNUST), which is in close proximity to the community.
3. Materials and methods
3.1. Earthworms
The earthworms Eisenia fetida and Eudrilus eugeniae were obtained
from Green Cycle Technologies Limited in Accra, Ghana. The species
were verified at the Biology Department, KNUST, and cultured in the
EQE laboratory. The earthworms were cultured in a plastic container
containing garden soil for one month. The vermibeds were kept moist
during culturing through regular sprinkling of water on top of the bed.
The top of the container was covered in order to create a dark atmo­
sphere and also prevent escape of the earthworms.
3.2. Collection of human excreta
The toilet facility had twelve (12) cubicles, six (6) cubicles each for
males and females. Each cubicle had containers for disposal of anal
cleansing material; hence these were not dropped into the containment
structure. In order to ensure collection of fresh excreta, uncontaminated
by toxic cleaning chemicals, six (6) samplers were designed to directly
receive the human excreta from three (3) male and three (3) female
cubicles. These samplers were removed whenever the facility was to be
cleaned to prevent possible contamination. Human excreta collected
were thoroughly mixed before analysis and feeding of experimental
setups.
3.3. Experimental setup
Factorial design experiment, involving two species of earthworms in
vermibeds and the co-decomposition of excreta and anal cleansing
material, was used to determine the sludge accumulation and mass
reduction as well as pollutants removal efficiencies by the treatment
units. The experiment employed three groups of design, EE and EF, EEA
and EFA, and NW, to assess excreta degradation rates and sludge
accumulation rates and how anal cleansing materials affect these rates.
EE and EF assessed how species type would affect performance while the
control, NW, was to help evaluate how earthworms influence degrada­
tion and sludge accumulation. For the loading rate, an average excreta
generation rate of 300 g/user/day was used. Experimental setups were
therefore designed to investigate a loading rate of 0.28 kg feed/m2/day.
This loading rate was determined based on 1 user/day and to achieve
this, the experimental setups were fed with 13 g of human excreta daily.
In order to estimate the amount of toilet paper used per person per day, a
simple survey of individuals (n = 30) on the length and weight of toilet
paper used per toilet visit was conducted and found to be 6 g/user/day,
corresponding to a loading rate of 0.006 kg feed/m2/day, for a digester
M.N. Acquah et al.
with surface area of 1.08 m2. To achieve this loading for the experi­
mental setups with surface area of 0.05 m2, 0.3 g of toilet paper was
added. Setups with surface area of 0.05 m2 were stocked with 15 worms
corresponding to a field density of 300 worms per m2.
Experimental setups consisted of two transparent containers (length
0.27 m, breadth 0.17 m and depth 0.12 m) stacked on top of each other,
allowing a small gap in between. The upper container, serving as the
vermibed, was perforated at the bottom to allow for drainage of effluent
and subsequent collection in the bottom container (Figure S2). Vermi­
beds consisted of a layer of soil on top of which was placed 200 g of coir
and a plastic mesh. Soil and coir served as supporting and bulking ma­
terials respectively. In addition to enriching the diversity of microbial
population and enzymatic activities, bulking material was needed to
improve the physical environment for support in the vermibeds. The soil
was collected from an agricultural field, below 10 cm of the surface, and
was properly homogenized before placing in the vermibeds. Charac­
teristics of the soil are shown in Table S1.
The vermibeds were covered with perforated lids to provide proper
air ventilation and also allow exchange of gases. The lids also prevented
escape of the earthworms. Twenty setups were prepared, each having a
duplicate. Two sets of experiments were run simultaneously. One was to
determine the degradation and accumulation rates of the fresh excreta
and the other the effect of toilet paper, as anal cleansing material, on the
rate of degradation and accumulation of sludge. Fifteen earthworms of
each species having individual live weight of 120.21 ± 2.81 mg for
Eisenia fetida and 202.46 ± 4.92 mg for Eudrilus eugeniae were released
into each experimental container (Table 1).
Setups EE, EF and NW were each fed with 13 g of human excreta
daily. Setups EEA and EFA were fed 0.3 g of anal cleansing material in
addition to 13 g of excreta daily. Approximately 22 ml of water was
added to each setup daily with the feed to mimic the microflush toilet
system (500 ml to 1 L per flush). The plastic meshes on which the excreta
were placed were weighed daily in order to determine sludge accumu­
lation rates.
Percentage mass reduction was calculated using Equation (1):
[(TFMA – FMR)/TFMA)] × 100
Where; TFMA = total faecal mass added; FMR = faecal mass remaining.
3.4. Physico-chemical and biological analyses of samples
In all the systems, moisture content, pH, electrical conductivity,
volatile solids, total organic carbon content, ash content, chemical ox­
ygen demand (COD) and nitrogen content were determined every week
during vermicomposting for 28 days. Once the samples were collected
analyses were performed immediately or the samples were kept in a
refrigerator at 4 ◦ C until analysis.
Moisture and ash contents were measured by using a gravimetric
method at 105 ◦ C within 24 h and 550 ◦ C for 4 h, respectively. pH and
electrical conductivity were measured using a digital pH meter and an
electrical conductivity meter, respectively. Volatile solids (VS) were
determined as sample weight loss (previously oven-dried at 105 ◦ C)
upon ashing at 550 ◦ C for 2 h in a muffle furnace. Total organic carbon
was calculated by multiplying the VS values by 1.8 (Rostami et al., 2010;
Wu et al., 2000). TKN was measured in accordance with the
Table 1
Description of experimental setup.
Code Description of setup
RW Fresh human excreta
EE Eudrilus eugeniae with excreta only
EEA Eudrilus eugeniae with excreta and toilet paper
EF Eisenia fetida with excreta only
EFA Eisenia fetida with excreta and toilet paper
NW Bed without earthworms (control setup)
Journal of Environmental Management 293 (2021) 112817
micro-Kjeldhal method (Bremner, 1996; Yadav and Garg, 2011). COD
analyses were done using the closed reflux titrimetric method
(APHA/AWWA/WEF, 2005). Helminth egg was measured using
Kato-Katz technique (WHO, 2019).
3.5. Statistical analysis
One-way analysis of variance (ANOVA) was conducted to determine
whether the mean difference of results from various systems were sta­
tistically significant. A Post-hoc LSD test was further conducted and
variations or differences were considered significant when (p ≤ 0.05)
(Montgomery and Runger, 2007).
4. Results and discussion
The characteristics of the fresh human excreta are presented in
Table S2. The results obtained showed that the human excreta had
average pH, conductivity, moisture content and total solids of 6.59,
1067.69 μS/cm, 78.51% and 21.49% respectively. Volatile solids, ash
content, carbon, nitrogen and COD concentration also recorded 83.92%,
16.08%, 48.79%, 9.92% and 167,918.75 mg/L respectively. For hel­
minth eggs analysis, the results obtained for Ascaris lumbricoides
(roundworm) and Ancylostoma duodenale (hookworm) in human excreta
were 305 ± 121.40 and 369 ± 49.58 eggs/g respectively. Analysis of
Trichuris trichiura (whipworm), Strongyloides stercoralis (threadworm)
and Schistosoma haematobium (blood fluke) in 1 g of human excreta
samples showed mean and standard deviation of 288 ± 83.14, 285 ±
65.69, 285 ± 65.69 and 156 ± 49.68 respectively (Figure S3).
A study on human excreta from a non-flush, drop and store type of
toilet from a village named Mandhana near IIT Kanpur, India reported
an average pH of 5.4, moisture content of 80%, total nitrogen of 4.03%
and total carbon of 42% (Yadav et al., 2011a). Another study by Nguyễn
(2012) in Germany, also documented an average pH value of 6.45, total
solids of 16.6%, total nitrogen of 2.5% and carbon of 50.25%. Several
researches have proven that the composition of human excreta varies
from geographical location due to factors such as prevailing environ­
(1)
mental conditions and type of diet (Rose et al., 2015).
4.1. Temperature in experimental setups during vermicomposting
Temperatures measured during vermicomposting are illustrated in
Figure S4. All the systems exhibited temperature values within the range
of 25–32 ◦ C with ambient temperature recording values between 32 and
38 ◦ C. Temperature in all vermibeds followed the trend of the ambient
temperature. Similar finding was made by Furlong et al. (2014a) where
the average temperatures in the vermifilters mirrored the patterns of the
average environmental temperatures. A study to monitor the life cycles
and optimal conditions for survival and growth of Eisenia fetida, Den­
drobaena veneta, Eudrilus eugeniae, and Perionynx excavates revealed that
each earthworm species varied substantially in terms of their tolerance
to varying temperatures. The suitable temperature range for Eisenia
fetida was between 0 ◦ C and 35 ◦ C while optimum temperature was
25 ◦ C. Eudrilus eugeniae and P. excavatus died at temperatures below 9 ◦ C
and above 30 ◦ C but had ideal temperature of 25 ◦ C (Domiguez and
Edwards, 2010b). Lower temperature recorded in various systems (NW,
EE, EEA, EF, EFA) than ambient temperature (ATM) might be due to the
daily watering of the systems that kept the temperature favourable for
the survival of the earthworms (Furlong et al., 2014b).
4.2. Degradation and accumulation of sludge during vermicomposting
4.2.1. pH
Figure S5, shows the changes in pH during vermicomposting. There
was an increase in pH from 7.75 ± 0.07 to 8.70 ± 0.05 from the first to
third week then a decrease to 7.82 ± 0.23 at the fourth week in EE
(Eudrilus eugeniae with excreta). Similar observation was made in EF
3
M.N. Acquah et al.
(Eisenia fetida with excreta) with a pH increase from 7.53 ± 0.17 to 8.56
± 0.39 at the end of the third week then a decrease to 7.75 ± 0.38 at the
end of the experiment. The rise in pH found in EE and EF during the first
21 days of vermicomposting may be attributed to carbonate release due
to urea hydrolysis (Khan et al., 2019). The decrease in pH recorded in
the fourth week in EE and EF could be attributed to the production of
CO2 due to the metabolic activities of earthworms and microorganisms
(Raphael and Velmourougane, 2011). In comparison to the systems with
earthworms (EE and EF), the control without earthworms (NW) recor­
ded an initial pH decrease up to day 7, reaching values as low as 5.62 ±
0.06. This could be attributed to anoxic conditions that favoured organic
acid accumulation (Buzie-Fru, 2010). This was not the case in the ver­
mibeds (EE and EF) presumably due to the aerating effects of earth­
worms. Statistically there was significant difference in pH between EE,
EF and NW (p = 2.17E-09) employing one-way ANOVA test which
means that there is variation in pH change in setups with and without
earthworms. Further LSD test conducted on the samples indicated that
there was significant variation between EE and NW (p = 0.000196) and
between EF and NW (p = 0.000196). Hence this may indicate that
earthworms play a major role in the change of pH of substrate during
vermicomposting.
There was a general decrease in pH at the end of the experiment in
both vermibeds, with and without toilet paper. EEA (Eudrilus eugeniae
with excreta and toilet paper) recorded a decrease in pH from 7.73 ±
0.04 in the first week to 6.67 ± 0.07 in the fourth week whiles EFA
(Eisenia fetida with excreta and toilet paper) decreased from 7.82 ± 0.03
to 6.75 ± 0.20. Generally there was no significant difference in the mean
pH (p = 0.114) in all vermibeds (EE, EF, EFA and EEA) using one-way
ANOVA. This may indicate that the addition of toilet paper does not
cause a difference in pH in various vermibeds over the experimental
period when Eisenia fetida and Eudrilus eugeniae were employed.
4.2.2. Electrical conductivity
Figure S6, shows the changes in electrical conductivity during ver­
micomposting. Electrical conductivity increased gradually in both the
system without earthworms (NW) and those with earthworms (EE and
EF) until day 14 when a decrease was seen in NW and then a slight in­
crease to day 28. There was a general decrease in EC in NW from 1099 ±
6.00 μs/cm in the first week to 736.5 ± 13.50 μs/cm at the end of the
fourth week. Moreover, there was an increase in EC in EE and EF from
1146 ± 46.00 μs/cm and 1099.5 ± 4.50 μs/cm in the first week to 1590
± 85.00 μs/cm and 1437.5 ± 32.50 μs/cm at the end of the fourth week
respectively. The rise in electrical conductivity could be attributed to the
loss of organic matter and the release of various inorganic ions (such as
phosphate, ammonium and potassium) (Garg et al., 2007). The electrical
conductivity variations were significantly different (p = 0.000111) in all
the setups (EE, EF and NW) using one-way ANOVA. Further LSD test
conducted indicated that there was significant variation between EE
(Eudrilus eugeniae with excreta) and NW (system without earthworms)
(p = 0.0000045) and between EF and NW (p = 0.001). This may indicate
that the various earthworms employed altered or played a significant
role in the change in electrical conductivity of the sludge.
One-way ANOVA showed no significant difference in mean EC (p =
0.423) in the various vermibeds (EE, EF, EFA and EEA). This demon­
strates that the addition of toilet roll did not cause a significant change in
EC value over the experimental period when both species of earthworms
were employed.
4.2.3. Moisture content
The percentage changes in moisture content in all setups were in the
range of 52–74% (Figure S7). The moisture content recorded during the
experiment fell within the optimum range for most earthworm species,
that is, between 50% and 90% (Dominguez and Edwards, 2010b). The
moisture content variations were not statistically different (p = 0.687)
in all the systems (EE, EF and NW) using one-way ANOVA.
The moisture content in vermibeds (EE, EF, EFA and EEA) fell within
4
Journal of Environmental Management 293 (2021) 112817
the ranges of 54%–76%. The highest moisture content of 75.11 ± 1.04%
was recorded in EFA (Eisenia fetida with excreta and toilet paper) in the
first week then decreased to 58.10 ± 0.22% in the fourth week. In EEA
(Eudrilus eugeniae with excreta and toilet paper) moisture content
reduced from 68.59 ± 0.56% in the first week to 54.22 ± 0.65% at the
end of the experiment (week 4). Statistical analysis employing one-way
ANOVA showed no significant difference (p = 0.233) in moisture con­
tent in setups (EE, EF, EFA and EEA). Conversely, the weekly moisture
content reductions were statistically different (p = 8.12E-09).
4.2.4. Total organic carbon
The general decrease of total organic carbon indicates mineralization
of organic matter (Fig. 1). The carbon contents of the biosolids for this
study were between 36 and 47% in EE and EF. Greater reduction was
observed in EF followed by EE than there was in the setup without
earthworm (NW) which indicates that earthworm activity greatly lowers
the levels of organic carbon in waste and speeds up the process of sta­
bilization (Suther, 2007). A fraction of TOC is lost as CO2 during ver­
micomposting owing to the consumption of the available carbon as a
source of energy by the earthworms and microbes causing a decrease in
the TOC value of biosolids (Sonowal et al., 2014a). The total organic
carbon variations were significant (p = 3.73E-08) in all the systems (EE,
EF and NW). Further LSD test conducted on the results indicated that
there was significant variation between EE and NW (p = 0.001), EF and
NW (p = 7.70E-09) and EE and EF (p = 0.000023).
In assessing the effect of toilet paper on the degradation in systems,
(EE and EEA) and (EF and EFA), there was a general decrease in carbon
in all vermibeds. An ANOVA test on the results showed significant dif­
ference (p = 0.012) between the vermibeds. A LSD Post-hoc test was
employed to compare the level of significance between the vermibeds
and there was significant difference between EE (Eudrilus eugeniae with
excreta) and EEA (Eudrilus eugeniae with excreta and toilet paper) (p =
0.026). There was no significant difference when EF (Eisenia fetida with
excreta) was compared with EFA (Eisenia fetida with excreta and toilet
paper) (p = 0.820). This indicates that the addition of toilet paper did
not influence carbon reduction when Eisenia fetida species of earth­
worms were employed.
4.2.5. Volatile solids and ash content
This study recorded 12.3% and 26.2% reduction in volatile solids in
EE and EF respectively. NW however recorded only 2% reduction in
volatile solids (Fig. 2a). The decline in VS values in EE and EFmay be
ascribed to quicker breakdown of organic content in sludge as a
consequence of raising its surface region with earthworm burrowing
operations. This phenomenon is consistent with Cofie et al. (2009) who
reported similar observations. The reductions in systems without
earthworms (NW) remained almost steady through the experimental
Fig. 1. Variation of Total organic carbon during vermicomposting (mean ±
standard error). For each treatment system independently, columns carrying
different superscript are significantly different (LSD test, p < 0.05).
M.N. Acquah et al.
Fig. 2. (a) Variation of Volatile solids and (b) Variation of Ash content during
vermicomposting (mean ± standard error). For each treatment system inde­
pendently, columns carrying different superscript are significantly different
(LSD test, p < 0.05).
period. The higher reduction over time found in EE and EF indicates that
earthworms were involved in the decomposition process and accelerated
the decrease of VS. Cardoso and Ramírez (2002) reported decreases in
VS ranging between 12% and 15%. Contreras-Ramos et al. (2005) also
reported a decrease in VS ranging from 13% to 22% during vermi­
composting of biosolids with cow manure and oat straw after 2 months,
using Eisenia fetida. Using one-way ANOVA there was significant dif­
ference (p = 3.74E-08) in all the setups (EE, EF and NW). Further LSD
test conducted on the results indicated that there was significant vari­
ation between EE and NW (p = 0.001), EF and NW (p = 7.73E-09) and
EE and EF (p = 0.000023).
The mean VS% was observed to decrease in all vermibeds with EEA
(Eudrilus eugeniae with excreta and toilet paper) recording the highest
removal at the 4th week. EFA (Eisenia fetida with excreta and toilet
paper) was observed to have slightly higher VS% reduction than EF
(Eisenia fetida with excreta). A one-way ANOVA showed a significant
difference (p = 0.012) in volatile solids between the various vermibeds.
LSD test further conducted indicated a significant variation between EE
(Eudrilus eugeniae with excreta only) and EEA (Eudrilus eugeniae with
excreta and toilet paper) (p = 0.026). Again there was significant dif­
ference (p = 0.005) between EE (Eudrilus eugeniae with excreta) and EFA
(Eisenia fetida with excreta and toilet paper). There was however no
significant difference when EF (Eisenia fetida with excreta) was
compared with EFA (Eisenia fetida with excreta and toilet paper) (p =
0.820).
The ash content in systems EE and EF increased till the fourth week
with almost similar trend with just a slight decrease during the first week
in the system without earthworms (NW) (Fig. 2b). The higher level of
ash content is a good measure of human excreta degradation (Sonowal
et al., 2014b). The ash content variations were significant (p =
0.000023) in systems EE, EF and NW using one-way ANOVA. Further
5
Journal of Environmental Management 293 (2021) 112817
LSD test conducted on the results indicated that there was significant
variation between EE and NW (p = 0.001), EF and NW (p = 7.70E-09)
and EE and EF (p = 0.000023).
In assessing the effect of toilet paper on the variation of ash content
over the experimental period, it was observed that there was an increase
in ash content in all vermibeds (EE, EF, EEA and EFA) at the end of the
vermicomposting period. A one-way ANOVA test on the results showed
significant difference (p = 0.012) in ash content in various vermibeds. A
Post-hoc LSD test indicated significant difference between EE (Eudrilus
eugeniae with excreta) and EEA (Eudrilus eugeniae with excreta and toilet
paper) (p = 0.026). There was no significant difference when EF (Eisenia
fetida with excreta) was compared with EFA (Eisenia fetida with excreta
and toilet paper) (p = 0.820). This may suggest that the addition of toilet
paper did not cause difference in ash content when Eisenia fetida was
employed. There was however significance difference between EE and
EFA (p = 0.005).
4.2.6. Total Kjeldahl Nitrogen
The result of the TKN variation during the experimental period is
presented in Fig. 3. There was a general decrease in nitrogen content
over the experimental period in EE and EF. The decrease in nitrogen
content in EE and EF can be related to the breakdown of organic com­
pounds with the influence of earthworms (Owusu-Antwi et al., 2017).
Buzie-Fru (2010b), observed that pH above 8.4 induced the change of
ammonium ion to ammonia gas which resulted in decrease in nitrogen in
substrate. The pH in this study were observed to be 8.70 ± 0.05 and 8.56
± 0.38 in EE and EF respectively at the third week. TKN variation were
significant (p = 0.015) in systems EE, EF and NW using one-way
ANOVA. Further LSD test conducted on the results indicated that there
was significant difference between EE and NW (p = 0.006) and EF and
NW (p = 0.025).
There was a decrease in nitrogen from the first week to the second
week in vermibeds with excreta and toilet paper (EEA and EFA) but a
gradual increase in nitrogen content in the third week was observed in
EFA (Eisenia fetida with excreta and toilet paper). A one-way ANOVA
showed no significant difference (p = 0.310) in nitrogen content be­
tween vermibeds with excreta and toilet paper (EEA and EFA) and with
excreta only (EE and EF). This indicates that the addition of toilet paper
did not influence nitrogen content when both species of earthworms
were employed.
4.2.7. Chemical oxygen demand
There was decrease in COD in vermibeds EE and EF throughout the
experimental period with the exception of NW which saw a slight in­
crease in COD in the second week then a gradual decrease till the fourth
week (Fig. 4). This reduction in COD concentration over the experi­
mental period was due to the fact that earthworms secrete enzymes that
Fig. 3. Variation of Total Kjeldahl Nitrogen during vermicomposting (mean ±
standard error). For each treatment system independently, columns carrying
different superscript are significantly different (LSD test, p < 0.05).
M.N. Acquah et al.
Fig. 4. Variation of Chemical Oxygen Demand during vermicomposting (mean
± standard error). For each treatment system independently, columns carrying
different superscript are significantly different (LSD test, p < 0.05).
help in the degradation of several other chemicals which cannot be
decomposed by microbes (Garkal, 2015). The COD variations were
significant (p = 0.011) in setups EE, EF and NW using one-way ANOVA.
Further LSD test conducted on the results indicated that there was sig­
nificant difference between EE and NW (p = 0.008) and EF and NW (p =
0.009) which suggest that both earthworms had significant impact on
the reduction of COD concentrations.
A one-way ANOVA conducted showed no significant difference in
COD between the various vermibeds testing the effects of the presence of
toilet paper (EE, EF, EEA and EFA) (p = 0.693). This suggests that the
addition of toilet paper did not influence the reduction of COD con­
centration in the sludge.
4.3. Helminth egg variation during vermicomposting
There was a general reduction in the various helminth eggs con­
centrations in vermibeds (EE, EF, EEA and EFA) at the end of 28 days of
vermicomposting (Figure S8 – Figure S12). Even though general
reduction was recorded, minimal increases in helminth eggs concen­
trations was observed within the first 2 weeks of vermicomposting.
Nuesca et al. (2007) also reported initial average count (mean) of
hookworm ova decreasing after 30 days of vermicomposting and
thereafter showing further increase after 60 days.
4.3.1. Ascaris lumbricoides
The average concentration of Ascaris in the feed was 305 ± 42.92
eggs/g. In EE, the concentration of Ascaris eggs decreased gradually over
the 4 weeks of vermicomposting (Figures S8). In NW, there was increase
in Ascaris egg concentration to the second week then a gradual decrease
to the fourth week. Despite the decrease found in the concentration of
the Ascaris eggs in the fourth week in the NW treatment, the concen­
tration was still higher than it was observed at the end of the first week.
Average Ascaris concentration of 168 ± 0.00 eggs/g, 120 ± 24 eggs/g,
408 ± 24 egg/g was recorded for EE, EF and NW at the end the exper­
iment (Fig. 2). There was statistically significant difference (p =
0.000169) in the Ascaris concentrations between systems with and
without earthworms. Further LSD test conducted on the results indicated
that there was significant variation between EE and NW (p = 0.000196)
and between EF and NW (p = 0.000196).
At the end of the fourth week Ascaris egg decreased to 120 ± 24.00
and 48 ± 0.00 in (EEA and EFA) respectively. However, a one-way
ANOVA showed no significant variation between the various setups
when toilet paper were added (EE, EEA, EF, EFA) (p = 0.512). The
percentage Ascaris egg reduction at the end of experimental period were
44.92%, 60.66%, 60.66% and 84.26% in EE, EF, EEA and EFA
respectively.
6
Journal of Environmental Management 293 (2021) 112817
4.3.2. Trichuris trichiura
There was a gradual decrease in Trichuris eggs throughout the
experimental period in EE and EF. Mean concentration of Trichuris eggs
in the feed throughout the experiment was 288 ± 29.39 eggs/g. Trichuris
concentration remained fairly constant in NW till after the third week
when a gradual decrease was observed to occur (Figure S9). The con­
centration of Trichuris decreased to 72 ± 24 eggs/g at the end of ver­
micomposting in both EE and EF. One-way ANOVA indicated a
significant variation between the samples (EE, EF, NW) (p = 0.006).
Further LSD test conducted on samples indicated a significant difference
between EE and NW (p = 0.031) as well as between EF and NW (p =
0.002).
At the end of the fourth week Trichuris eggs decreased to 24 ± 0.00 in
both EEA and EFA. However a one-way ANOVA showed no significant
variation (p = 0.453) between the various vermibeds (EE, EF, EFA,
EEA). This may indicate that the addition of toilet roll did not cause any
significant difference in Trichuris removal over the experimental period
when both species of earthworms were employed.
4.3.3. Ancylostoma duodenale
The initial average Ancylostoma egg concentration for this study was
369 ± 17.53 eggs/g. It decreased to 120 ± 24 and 96 ± 24 eggs/g in EF
and EE respectively. In NW, the Ancylostoma eggs decreased from the
first to the second week. An increase in number was however observed
in the third week before a further decrease to the fourth week
(Figure S10). The percentage Ancylostoma eggs concentration reduction
at the end of experiment in EE, EF and NW were 67.5%, 74% and 41.5%
respectively. A one-way ANOVA indicated a significant variation be­
tween the samples (p = 0.01). Further LSD test conducted on the results
indicated that there was significant difference between EE and NW (p =
0.006) and between EF and NW (p = 0.000337).
At the end of the fourth week Ancylostoma eggs decreased to 48 ±
24.00 and 24 ± 0.00 in EEA and EFA respectively. However the per­
centage Ancylostoma egg removals EE and EEA after the same duration
of experimentation were 67.48% and 86.99% respectively. 73.98% and
93.51% reduction in Ancylostoma egg concentration were observed in EF
and EFA respectively. A one-way ANOVA on the results showed no
significant variation (p = 0.512) between the various systems (EE, EEA,
EF, EFA) when toilet paper was added to assess its effect.
4.3.4. Strongyloides stercoralis
The mean level of Strongyloides concentration applied as feed into
various vermibeds was 285 ± 23.32 eggs/g. EE performed well in the
reduction of strongyloides concentration at the end of the experiment
with 58% removal. Generally, there was a decrease in Strongyloides eggs
concentration over the four weeks of vermicomposting in EE and EF. In
NW, the Strongyloides egg concentration decreased from the first to the
second week. An increase in concentration was however observed in the
third week before a further decrease to the fourth week (Figure S11).
There was significant difference between EE, EF and NW (p = 0.009)
using one-way ANOVA. Further LSD test conducted on the results indi­
cated that there was significant difference between EE and NW (p =
0.005) and between EF and NW (p = 0.013).
At the end of the fourth week Strongyloides eggs decreased to 120 ±
24.00 and 96 ± 24.00 in EEA and EFA respectively. However a one-way
ANOVA showed no significant variation between the systems (EE, EF,
EFA, EEA) (p = 0.955). The percentage Strongyloides eggs reduction in
EEA and EFA at the end of experimental duration were 57.89% and
66.32% respectively.
4.3.5. Schistosoma haematobium
There was a gradual decrease of Schistosoma eggs concentration from
a mean initial concentration of 156 ± 17.57 eggs/g to 24 ± 0.00 eggs/g
and 48 ± 24.00 eggs/g over the 4 weeks of vermicomposting in EE and
EF respectively (Figure S12). There was an increase in Schistosoma eggs
concentration to the second week then a decrease to the fourth week in
M.N. Acquah et al.
NW. A one-way ANOVA indicated a significant variation between the
samples (EE, EF and NW) (p = 0.004). Further LSD test indicated that
there was significant variation between EF and NW (p = 0.002) and
between EE and NW (p = 0.005).
At the end of the fourth week Schistosoma eggs decreased to 24 ±
0.00 in both (EEA and EFA) respectively. However there was no sig­
nificant difference between the vermibeds (EF, EFA, EE, EEA) (p =
0.973) using one-way ANOVA. This may indicate that the addition of
toilet roll did not cause any difference in Schistosoma concentration
reduction.
4.4. Mass reduction and accumulation of sludge
4.4.1. Mass reduction
Fig. 5, shows the cumulative mass reduction in systems with and
without anal cleansing materials. CMA represents the cumulative mass
of feed added to the various systems. In systems without anal cleansing
material (EE, EF and NW), it was observed that mass reduction was
higher in systems with earthworms (EE and EF) than in systems without
earthworms (NW). This reduction could be attributed to the activities of
earthworms which aided in the rate of degradation and reduction of the
human excreta (Furlong et al., 2014c). The mass reduction recorded at
the end of the fourth week in EE, EF and NW were 67.5% (245.85 g),
58.8% (214.18 g) and 40.5% (147.49 g) respectively. Again 73.7%
(274.38 g) and 68.5% (255.12 g) was recorded at the end of the fourth
week in EEA and EFA respectively. There was significant variation over
the weeks of vermicomposting (p = 0.001) using one-way ANOVA,
confirming that both species of worms were actively degrading human
excreta with respect to time.
4.4.2. Sludge accumulation
Fig. 6, shows the volume of sludge accumulated in the various setups
Fig. 5. Cumulative mass reduction in experimental setup (a) NW, EE, EF, and
(b) NW, EEA and EFA during vermicomposting (mean ± standard error).
7
Journal of Environmental Management 293 (2021) 112817
Fig. 6. Accumulation of sludge during vermicomposting (mean ± standard
error). For each treatment system independently, columns carrying different
superscript are significantly different (LSD test, p < 0.05).
over four weeks using density of faeces, 1070 kg/m3 (Penn et al., 2018).
Quantities of sludge accumulated at the end of the experimental dura­
tion were 0.00020 m3, 0.00011 m3, 0.00014 m3 in NW, EE, EF whilst
EEA and EFA recorded 0.00009 m3 and 0.00011 m3 respectively. The
relatively lower accumulation in EE and EF than NW could be attributed
to the earthworms contributing to the sludge reduction process via
consumption of the human excreta and also aiding the rate of degra­
dation (Yadav et al., 2011b). There was significant difference (p =
0.0000056) in the accumulation across the different setups over the
weeks (EE, EF and NW) using one-way ANOVA. This indicates that the
accumulation of sludge decreases in each experimental setup due to the
actions of the earthworms with respect to time.
In assessing the effect of anal cleansing material, a one-way ANOVA
indicated significant difference (p = 1.69E-10) in accumulation of
sludge in the various vermibeds (EE, EF, EEA and EFA) over the weeks.
This indicates that addition of toilet paper influences sludge accumu­
lation with respect to time.
5. Conclusion
The study revealed that earthworms significantly improved the
degradation and mass reduction of human excreta. The least mass
reduction was observed in the setup without earthworms (NW) 40.5%
(147.49 g), with setups having earthworms recording 67.5% (245.85 g)
and 58.8% (214.18 g) for EE (Eudrilus eugeniae and excreta only) and EF
(Eisenia fetida and excreta only) respectively. The mass reduction in
vermibeds (EE and EF) was relatively lower than vermibeds with excreta
and toilet paper (EEA and EFA). EEA (Eudrilus eugeniae with excreta and
toilet paper) and EFA (Eisenia fetida with excreta and toilet paper)
recorded a mass reduction of 73.7% (274.38 g) and 68.5% (255.12 g)
respectively. With respect to reduction of sludge in experimental setups
with and without toilet paper, it was found that Eudrilus eugeniae species
of earthworm was superior to Eisenia fetida. The addition of toilet paper
in the study did not result in any detrimental effects on the degradation
of excreta, neither did it increase sludge accumulation rates, as feared by
patrons of vermidigesters but rather resulted in lower rates of accumu­
lation. This research corroborates vermicomposting as an effective­
means for the degradation of human excreta together with anal
cleansing material (toilet paper). However, helminth egg can still be
present at potentially unsafe levels after 28 days of vermicomposting
hence would require further treatment and proper handling. The study
recommends that a pilot field test is performed to evaluate the real-time
effects of adding anal cleansing material (toilet paper) to vermidigesters.
Authors’ statement
1. Michael Nyame Acquah: Writing -Original Draft, Methodology,
Formal analysis, Investigation. 2. Helen Michelle Korkor Essandoh:
Conceptualization, Supervision, Writing- Reviewing and Editing,
M.N. Acquah et al.
Visualization. 3. Sampson Oduro-Kwarteng: Supervision, Writing-
Reviewing and Editing. 4. Eugene Appiah-Effah: Writing- Reviewing
and Editing. 5. Peter Owusu-Antwi: Writing- Reviewing and Editing.
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.
Acknowledgements
This study was funded by the Regional Water and Environmental
Sanitation Centre Kumasi (RWESCK) at the Kwame Nkrumah University
of Science and Technology (KNUST), with funding from the Ghana
Government, through the World Bank under the Africa Centres of
Excellence Project. The views expressed in this paper do not reflect those
of the World Bank, Ghana Government and KNUST.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.jenvman.2021.112817.
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