Desalination 282 (2011) 95103

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Desalination
journal homepage: www.elsevier.com/locate/desal

Urban wastewater treatment using vermi-bioltration system

Priyanka Tomar, Surindra Suthar
School of Environment & Natural Resources, Doon University, Dehradun-248001, India

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

Article history: This work illustrates the potential of a novel vermi-bioltration system in treatment of urban wastewater. A
Received 3 June 2011 small-scale vermi-bioltration reactor was constructed using vertical subsurface-ow constructed wetlands
Received in revised form 8 September 2011 (VSFCWs) aided with local earthworms Perionyx sansibaricus. The coco-grass: Cyprus rotundus (density 0.14
Accepted 9 September 2011
plants/in. 2) was used to construct VSFCW. Another reactor without earthworms acted as experimental con-
Available online 13 October 2011
trol. The wastewater was treated through this system for a total of eight repetitive cycles and after each cycle
Keywords:
the changes in pH, electrical conductivity (EC), total dissolved solids (TDS), and total suspended solids (TSS),
Wastewater treatment chemical oxygen demand (COD), NO3 and PO43 of water were measured. Vermi-bioltration caused signif-
Perionyx sansibaricus icant decrease in level of TSS (88.6%), TDS (99.8%), COD (90%), NO3 (92.7%) and PO43 (98.3%). There were
COD about 38.8, 20.8, 80.6, 50.8 and 144.6% more removal of TSS, TDS, NO3, PO43 and COD, respectively in
TDS vermi-bioltration than control. Results thus suggested that vermin-bioltration system is more efcient
Vermi-bioltration than VSFCW in terms of contamination removal efcacy. However, this work provides a preliminary idea of
Marshy plants using earthworms in wastewater treatment system and further detailed studies are required on some key issues
(e.g., loading rate, ow alternation impacts and earthworm stocking density) of this system.
2011 Elsevier B.V. All rights reserved.

1. Introduction in order to conserve the water and nutrient resources. An economical

The urban runoff in general, carries organic load along with sever-
al hazardous chemicals which not only spoils the aesthetic sense of
the river but at the same time also degrades the aquatic ecosystem.
Due to high establishment and running cost of a sewage treatment
plant (STP) the majority of urban centers in developing world dispose
urban runoff and sewerage water directly into urban river without
any treatments or with partial treatments. Several mechanical and
chemical approaches are being applied widely for urban wastewater
treatments systems in urban centers mainly by sewage treatment
plants (STPs). Apart to construction costs the operation and mainte-
nance problems in STPs has raised the question of sustainability [1].
Moreover, excess sewage sludge produced by STPs has been sub-
jected to increasingly stringent limitations on discharge during the
last few decades [2]. According to Sinha et al. [3] many developing
countries cannot afford the construction of STP and therefore; there
is growing concern over developing some ecologically safe and eco-
nomically viable small-scale wastewater treatment technologies for
onsite wastewater treatment. However, at this crucial juncture some
ecologically engineered tools can solve issues related with safe and
cost-effective wastewater treatments technologies. The majority of
present wastewater treatment systems are a disposal-based liner
system and they should be transformed into cyclical treatments [4]
Corresponding author. Tel.: + 91 135 2255103.
E-mail address: suthariitd@gmail.com (S. Suthar).
and manageable wastewater treatment approach is often required
and deserves to be explored [5].
Biological wastewater treatment process involves the potentials of
some living organisms to remove contaminants and sludge from
wastewater in order to make it suitable for surface irrigation and
other industrial use. Biological wastewater treatment involves the
transformation of dissolved and suspended organic contaminants to
biomass and evolved gases: CO2, CH4, N2 and SO2 [6]. A variety of or-
ganism like aquatic plants, marshland plants, protozoa, nematodes,
oligochaetes have been tested in both laboratory and eld conditions
to develop a low-cost bioreactor for wastewater treatment and sludge
reduction. The potential of oligochaetes for wastewater treatment
and sludge has been explored widely in many parts of the world. In
general, Oligochaetes can be divided into two distinct groups, rstly,
microdrilli (aquatic and small sized worms) and, secondly, terrestrial
oligochaetes (earthworms) [7]. The aquatic oligochaetes can be divid-
ed into two groups: (i) the large aquatic worms (Tubicidae, Lumbri-
culidae and the semi-aquatic or terrestrial Enchytraeidae) and, (ii) the
small aquatic worms such as Naidids and Aeolosomatids [8]. In recent
years, both aquatic and terrestrial oligochaetes have been tested by
several authors under lab-based trials to remove water contaminants
and excess quantity of sludge [3, 6, 8-13]. The major components and
outcomes of previous experiments on vermi-bioltration are de-
scribed in Table 1.
The utilization of earthworms in wastewater or sludge treatment
is called vermi-bioltration. It was rst advocated by the Prof. Jose
Toha at the University of Chile in 1992 [22]. Vermi-bioltration is a

0011-9164/$ see front matter 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.desal.2011.09.007
96 P. Tomar, S. Suthar / Desalination 282 (2011) 95103

Table 1
Earlier studies indicating potential of oligochaetes in sludge stabilization and wastewater treatment.

Type of sludge/wastewater Worm species used Major observations References

Aquatic worm ((Tubicidae, Lumbriculidae, Naidids and Aeolosomatids)

Waste sludge produced in wastewater Lumbriculus variegatus Sludge reduction was 77% during Hendrickx et al. [11]
treatment plant the process
Waste sludge produced in wastewater Lumbriculus variegatus TSS reduced up to 99% after Elissen et al. [14]
treatment plant treatment
Efuent of activated sludge process Lumbriculida hoffmeisteri Worm effectively removes major Wei et al. [15]
pollutant from sludge
Sludge from wastewater water Branchnria Sowerbyi, Limnodrilns There was drastic impact on sludge Song et al. [16]
(46.4% reduction in rst stage),
COD (more than 80% removal) and
BOD (more than 81%) during the process
Sewage of domestic sludge Lumbriculus variegatus 2040% sludge converted into worm Buys et al. [17]
biomass and nitrate as well as nitrite
removed efciently

Earthworms
Domestic wastewater (assessment of toxicity of Eisenia fetida High salt concentration may cause Hughes et al. [10]
ammonia on earthworm in vermi-bioltration system) damage to earthworms in vermitration
units
Liquid waste products from dairy industry E. fetida Removal of 5 day BOD by 98%, Sinha et al. [3]
COD by 8090%, TDS by 9092%
during the process
Domestic wastewater sludge E. fetida Signicant reduction in pollutant Wang et al. [5]
during vermistabilization process
Secondary liquid efuents from Gelatine Industry Lumbricus rubellus Decrease in COD by 90% and BOD Ghatnekar et al. [18]
by 89%.
Raw sewage E. fetida, Perionyx excavatus, Removal of COD by 8090% and Sinha et al. [19]
Eudrilus euginae BOD by 90% during vermi-bioltration
Treatment of sewerage and sludge E. fetida Removal of COD by 8186% and Xing et al. [20]
BOD by 9098% during vermi-bioltration
Domestic wastewater treatment E. fetida Removal of COD by 5566% and Xing et al. [21]
BOD by 4765% during process

process that adapts traditional vermicomposting system into a pas-
sive wastewater treatment process by using potentials of epigeic
earthworms. According to Komarowski [23] in vermi-bioltration
system suspended solids are trapped on top of the vermilter and
processed by the earthworms and fed to the soil microbes immobi-
lized in the vermilter. The dissolved and suspended organic and in-
organic solids are trapped by adsorption and stabilization through
complex biodegradation processes that take place in the living soil
inhabited by earthworm and the aerobic microbes. Intensication of
soil processes and aeration by earthworms enable the soil stabiliza-
tion and ltration system become effective and smaller in size [19].
In general, inoculated earthworms in vermibeds accumulate many or-
ganic pollutants from the surrounding soil environment, passive absorp-
tion through the body wall and also intestinal uptake during the passage
of soil through the gut [24]. The efcacy of vermi-bioltration system is
already described in literature (Table 1). Sinha and his group investigated
the potential of vermi-bioltration system in treatment of dairy industry
efuent [3]. They claimed that earthworms have been found to remove
the 5 day BOD by over 98%, COD by 8090%, TDS by 9092% from any
liquid wastes by the general mechanism of ingestion and biodegrada-
tion of organic wastes. According to a study conducted by Ghatnekar
et al. [18] suggested that the vermi-bioltration system is efcient to re-
move COD and BOD load of wastewater generated from gelatin indus-
try. They applied a three-tier biotechnology unit coupled with vermi-
bioltration system to convert secondary liquid efuents from a gelatin
manufacturing unit into bio-safe clean water. Results thus, suggested a
signicant decrease in COD by 90% and BOD by 89%. Recently, Zhao et al.
[2] studied the stabilization of domestic waste water sludge using
earthworms and results have revealed that the presence of earthworms
in the vermibeds to the signicant stabilization of the sludge. The vola-
tile suspended solids (VSS) reduction in the vermibeds was in the
ranges of 56.266.6% in different treatment units aided with earth-
worms. Similarly, Sinha et al. [19] developed a low-cost sustainable
technology over conventional systems to recycle the domestic waste-
water with potential for decentralization facility for waste manage-
ment. They claimed removal of 5 days' BOD (BOD5) by over 90%, COD
by 8090%, total dissolved solids (TDS) by 9092%, and the total sus-
pended solids (TSS) by 9095% from urban wastewater after the treat-
ment with worms. The microbes play an important role in vermi-
bioltration system and they also provide some extracellular enzymes
to facilitate the earthworms for rapid degradation of organic substances
in vermibeds [25]. Likewise, Zhao et al. [2] investigated the interactions
between microorganism and earthworm in vermi-bioltration system.
They demonstrated that earthworm biolm was dominated by the
members of the phylum Proteobacteria and Pseudomonas sp.
The majority of previous studies are available on either utilization
of vermi-bioltration or only constructed wetland ltration system
for removal of nutrients/pollutants from wastewaters, but no com-
prehensive report is available on utilizing potentials of both systems
to develop an effective integrated system, comprising of earthworm
and construction wetland system, for wastewater treatment. Al-
though, Chiarawatchai [26] has conducted an interesting study on
combining vertical sub-surface ow constructed wetlands (VSFCWs)
with earthworm. The integration of these two ecological techniques
(traditional wetlands system with vermi-bioltration mechanism)
can be a cost effective and sustainable option for onsite wastewater
treatment.
The aim of this study was to assess the potential of an integrated
vermi-bioltration system with VSFCWs constructed by using earth-
worm Perionyx sansibaricus and a wetland weed Cyprus rotundus
(coco-grass or red nut sedge) under a small-scale laboratory experiment.
C. rotundus is one of the most invasive weeds and have been reported
from tropical and temperate regions of the world. It is a perennial
plant and mainly occurs in gardens, agriculture plots, around stagnate
water bodies etc. Few earlier studies have demonstrated the capabilities
of C. rotundus in wastewater treatment and phytoremediation [2729].
 P. Tomar, S. Suthar / Desalination 282 (2011) 95103
C. rotundus was selected for vermi-bioltration system due its local and
perennial availability, short length (up to 40 cm) and easy cultivation
capabilities.
2. Materials and methods
2.1. Earthworm, plant collection and wastewater collection
Individuals of earthworm Perionyx sansibaricus of different age group
were collected from mud of a gray water drain in university campus,
Dehradun, India. The stock of P. sansibaricus was cultured in laboratory
using garden soil spiked with leaf litter and cow dung inappropriate ra-
tios. Plant Cyprus rotundus used for construction of bioltration unit
was originally obtained from moist soils around grey water drains in uni-
versity campus. The identication was made using standard taxonomic
key and conrmed by plant taxonomist in university.
The urban wastewater was collected from a wastewater stream
owing over nearby location of university campus. The wastewater
was collected from main streamline of wastewater drain in large-
size pre-cleaned circular plastic containers of 20 L capacity. Collected
wastewater was brought immediately to laboratory and collected in
large-size wastewater reservoir unit of the vermi-bioltration sys-
tem. Wastewater was collected just before the starting of experimen-
tation in order to avoid alternation in the wastewater characteristics
mainly due to open storage of sample.
Other accessories like aeration pump, ow control units, water
pumps etc. were procured from a local sanitary engineering shop
and scientic equipment supply rms.
2.2. Construction of vermi-bioltration and bioltration units
The experimental vermi-bioltration /bioltration units were com-
prised of two reactors/batches: (i) long cylindrical unit: Reactor-I and
(ii) rectangular unit: Reactor-II. Long unit was constructed with a tradi-
tional water ltration system using gravels and sand column at the base
and a bioltration system at the top made of living individuals of wetland
plant stand, i.e. Cyprus rotundus and earthworms in its root zone system.
An aeration unit was also xed in the middle layer of the Reactor-I. A plas-
tic circular cylinder of 80 L capacity was used to construct Reactor-I of
vermi-bioltration/bioltration unit. Following materials/layers were
used to ll (from bottom to top) the circular cylinder to construct the
vermi-bioltration/bioltration unit:
Layer I Large stones (1015 cm in diameter) up to 5-inch this
layer creates a kind of air chamber system and for water storage
in base of system.
Layer II Thick layer of small stones and gravel (57 cm diameter) up
to 2-inch acts as ltration unit and creates a kind of turbulence
during water ow and provides space for aeration of wastewater.
Aeration pipe (pierced 1 inch diameter and 15-inch length). Aera-
tion pipe was covered with 1 inch layer of small pebbles. A ne
plastic net was placed over the pebble layer aeration device
was installed in order to remove BOD load of the wastewater.
Layer III A thick layer of sawdust spread over the net (2-inch)
saw dust acts as good absorbent for several kinds of inorganic pol-
lutants of wastewater.
Layer IV Dried leaves of Sal tree were placed over sawdust layer
(2-inch) as natural adsorbent to remove nutrients from waste-
water. It also acts as feed for microbial communities helping in
wastewater mineralization.
Fine net The ne plastic net (b0.5 mm pore-size) was placed over the
leaf litter layer in order to check the entry of earthworm in deep
layers of the vermi-bioltration system in order to avoid moving
earthworm to deep bottom layers of the reactor.
97
Layer V Vermi-bioltration bed mainly constructed using thick
bedding of soil mixed with small stones and pebbles along with
complex root-zone system of surface plant Cyprus rotundus. The
thickness of this layer was about 10 in. earthworm acts as bio-
logical agent to remove solid fractions of wastewater and mineral-
ization of wastewater mainly driven by earthworm-microbe
interactions in root-zone system.
Layer VI Composed of surface vegetation stand of Cyprus. It was about
4 6 in. in length wetland plant provides air in root-zone system
and removes nutrients from wastewater through general absorption,
adsorption and translocation processes. Also provides shelters to
benecial microbial communities responsible for N mineralization.
The detail of vermi-bioltration/bioltration unit is given in Fig. 1.
In the top layer of Reactor-I, i.e. Layer-V the fresh and viable speci-
mens of Cyprus were planted in top soil layers. The roots of plant were
planted deeply and surface layer was irrigated regularly (for one
week) by tap water in order to x the planted Cyprus in top layer of ver-
mireactor. The mean density of Cyprus in vermireactor-I was about19
plants/in.2. The open space between plant stand was lled with a thin
layer of small stone to avoid direct hydraulic impact on the plant and
earthworm. In this vermi-bioltration system efforts were made to cre-
ate a kind of soil ecological system mainly comprised of thick soil layer
spiked with complex rooting system of Cyprus rotundus. The pieces of
stones and pebbles in this root-zone-ltration system create an appro-
priate space for air and inoculated earthworm in sub-soil system. None-
theless, the root-zone system not only enhances the efciency of
wastewater ltration but at the same time also provide shelters to bac-
terial communities (e.g., N-xers, ammonifying and denitrication bac-
teria) responsible for nutrient removal from wastewater.
Reactor-II: Another unit of reactor, i.e. Reactor-II was introduced in
order to enhance the removal efciency of the system. In Reactor-II
the biological component of the ltration unit was of more importance
therefore the majority of the reactor volume was lled with earth-
worm and plant root zone layers. A rectangular plastic container of
size (23. 5-inch length 18-inch width 15-inch depth) was used to
construct the second unit (Reactor-II) of vermin-ltration system.
In Reactor-II there were two district layers: rstly, base layer (large
pebbles; 1015 cm in diameter and height about 6-inches), and sec-
ondly, top layer (small pebbles; 57 cm in diameter mixed with ne
sand and height up to 10-inches). The top layer acts as bedding sub-
strate for earthworms in Reactor-II. A thin plastic net sheet was placed
between the both layers to avoid movement of earthworms from top
layers to base layer of the vermireactor. Fresh and viable specimens
of plant: Cyprus rotundus were planted in top layer and thin patches
of small stones were placed over the open spaces around Cyprus
plant to avoid direct hydraulic impact of inow water in Reactor-II
on plant stand and earthworms. The mean plant density in vermireac-
tor was 0.14 plants/in. 2 (calculated using values of total surface area
of Reactor-II and plant numbers in reactors). After plantation Cyprus
stand was allowed to grow for one week and during this period ade-
quate amount of tap water was supplied in vermireactor to facilitate
the xing of roots of plant in top layers of Reactor-II. Initially, both
reactors were run for twothree days using fresh tap water to
wash and x the layers of vermibeds in proper functioning forms.
After establishment of plant stands (after one week) the reactor
was run for wastewater treatment experimentations. In both exper-
imental vermi-bioltration systems, i.e. Reactor-I and Reactor-II indi-
viduals of earthworm P. sansibaricus were introduced over the
top layer the reactors. Small passages were made in the surface
layers of both reactors in order to facilitate worms to enter in the
top soil layers of the vermireactor. The initial earthworm density in
both vermi-bioltration systems was measured in the ranges of
22.024.5 g/L. The earthworms were allowed to settle in vermireac-
tors for initial 23 days and thereafter, vermireactor was run for
98 P. Tomar, S. Suthar / Desalination 282 (2011) 95103

Water Sprinkling Device

Supply of wastewater
from tank to reactor-I
through pump

Layer -VI

Layer - V

Layer - IV
Layer -III
Aeration pipe

Layer - II

Layer - I

Aerator
Reactor-I
Wastewater Tank
Outlet from Reactor-I

Layer -III

Layer -II

Layer -I

Reactor -II
Outlet (completion of cycle

Fig. 1. Vermi-bioltration system used for wastewater treatment.

experimentation. A reactor without earthworm (bioreactor) acted as of top layer, i.e. layer-V. The care was taken to avoid the overowing of
experimental control for this study. water. The wastewater was retained for 1 h in Reactor -I and a continu-
ous air was supplied during this period using an electronic aeration de-
2.3. Observation and data collection vice. After that the outlet of Reactor-I was opened into Reactor-II and

The wastewater was used without any dilation for this experimen- Table 2
tation. However, prior to putting wastewater in experimentation cycle Characteristics of wastewater used for experimentations.
a sample of wastewater (about 1 L) was separated from stock and an- Parameters Range
alyzed for its physic-chemical characteristics (Table 2). As illustrated
pH 7.61 0.10
in Fig. 1, during experimentation cycle the stock wastewater was sup-
EC (S/cm) 922.0 5.29
plied in Reactor-I through a mechanical pump and a ow control device TSS (mg/L) 216.67 7.64
was also xed in main water-supply pipe. The wastewater was sprinkled TDS (mg/L) 56813.3 51.3
over the surface of top layer of Reactor-I through a perforated plastic pipe NO3 (mg/L) 384.2 1.00
PO43 (mg/L) 36.37 0.67
and outlet of Reactor-I was closed to ll the reactor with wastewater. The
COD (mg/L) 863.0 3.60
wastewater was lled in reactor continuously up to the saturation level
 P. Tomar, S. Suthar / Desalination 282 (2011) 95103 99

ow of outlet was controlled using a mechanical ow control device. The
outlet water from Reactor-I was sprinkled over the surface of Reactor-II
using a perforated pipe system. In second cycle of treatment (i.e., treat-
ment in Reactor-II) the water retention mechanism and time framework
was same as used in Reactor-I. After that the outlet of Reactor-II was
opened to release the water from second reactor, i.e. Reactor-II. The com-
plete passing of water from both reactors was counted as one treatment
cycle and water after each cycle was putted back into new cycle. The
wastewater was repeatedly passed through both units of vermiltration
system for complete 8 cycles. An interval (stabilization period) of 24 h
was kept between two subsequent treatment cycles in order to stabilize
the microbial environment and earthworm population in sub-surface of
the vermireactor after each cycle. A sample of wastewater was collected
in pre-cleaned and sterilized polythene bottle of 1 L capacity from outlet
of Reactor-II after each treatment cycle and stored at 4 C for further in-
vestigations on changes in physico-chemical characteristics of waste-
water during each cycle.
2.4. Chemical analysis
The chemical characteristics of wastewater samples collected after
each treatment recycle were analyzed for different physic-chemical pa-
rameters by following methods as described by APHA-AWWA-WPFC
[30]. pH was measured using digital pH meter (Metrohm, Swiss-made).
Conductivity was measured using digital conductivity meter (Remi,
India). Total dissolved solids (TDS) and total suspended solids (TSS)
was measured was measured ltration and gravimetric and oven drying
methods. Chemical oxygen demand (COD) was measured using po-
tassium dichromate oxidation method. Nitrate, sulphate and phos-
phate contents in water were analysed spectrophometrically by
following methods as described by APHA-AWWA-WPFC [30].
2.5. Statistical analysis
A paired sample t-test between control (without earthworm) and
experimental (with earthworm) vermi-bioltration unit was per-
formed for each chemical parameter to analyze the differences.
One-way analysis of variance (ANOVA) was also preformed to mea-
sure the difference among different cycles for each physic-chemical
parameter of wastewater. SPSS statistical package (Window Version
13.0) was used for data analysis. All statements reported in this study
are at the p b 0.05 levels.
3. Results and discussion
The quality of wastewater in terms of phyico-chemical character-
istics is described in Table 2. The collected sample of urban wastewa-
ter showed relatively high values of some key pollution indicating
parameters of water: TDS (50813 mg/L), NO3 (384.2 mg/L), PO42
(36.37 mg/L), SO42 (293.3 mg/L) and COD (863.3 mg/L). The waste-
water after vermi-bioltration process showed a drastic change in
its major physico-chemical parameters, after each treatment cycle.
Although, there was signicant reduction in key pollutants of urban
water in both bioltration (without earthworm) and vermi-bioltration
(with earthworm), but difference was more prominent in water from
vermi-bioltration unit than initial levels. The changes in all reactors
pH Experiment
8.4 Control
8.2
8
7.8
7.6
7.4
7.2
7
6.8
0 1 2 3 4 5 6 7 8
Treatment cycle
Fig. 2. Changes in pH during different cycles of treatment in control and experimental
reactor.
7th cycle followed by pH stabilization state during last treatment
cycle. pH of water mainly depends upon a variety of chemical factors,
e.g., dissolved gases, organic acids, humic fractions and inorganic
salts. The decomposition of organic fractions of wastewater, mainly
by microbes in water, produces some acidic species of mineralized or-
ganic materials (CO2, ammonia, NO3 and organic acids) which plays
an important role in shifting of pH scale of treated water. Probably
the reduction in the level of ammonia, NO3 during bioltration and
vermi-bioltration treatment caused sight changes in pH. There was
statistically signicant difference between bioltration and vermi-
bioltration process for pH level (t-test: p b 0.05) of efuent from reac-
tors. The pH value of efuent obtained at the end of treatment process
was 7.81 in bioltration (about 9.1% more than initial) and 8.15 in
vermi-bioltration (about 13. 8% more than initial) reactor. The differ-
ent between control and experimental reactor for pH could be related
to earthworm mediated rapid mineralization of organic fractions of
wastewater. Also few earlier researchers have reported increase in
pH after vermi-bioltration processes [19, 21].
3.2. Electrical conductivity (EC)
Electrical conductivity (EC) of wastewater showed signicant changes
after treatment through ltration system in both bioltration and vermi-
bioltration processes. The conductivity of treated water was: 1230.0
S/cm in bioltration and 984.7 S/cm in vermi-bioltration system. The
difference between control and experimental reactor was statistically
signicant (t-test: p =0.002). The changes in EC during different cycles
are described in Fig. 3. In bioltration system EC showed a linear trend
of increase till last observation while in vermi-bioltration reactor EC
showed different patterns of uctuation during experimental processes.
In vermi-bioltration system EC value of efuent showed increasing
pattern up to 5th cycle of treatment thereafter, it reduced sharply till
last observation. The increasing EC could be attributed to mineralization
of organic waste fractions of wastewater through microbial and
EC Experiment
1400 Control
1300
1200
1100
S/cm

could be attributed to the development of biological communities with- 1000

in reactors [26].
900
800
3.1. pH
700
The change in pH during different treatment cycle is illustrated in 600
0 1 2 3 4 5 6 7 8
Fig. 2. In bioltration system (control) a trend of slight increment in
Treatment cycle
pH was observed till last observation. On the other hand, in vermi-
bioltration system pH decreased sharply up to 3 rd cycle of treat- Fig. 3. Changes in EC during different cycles of treatment in control and experimental
ment thereafter, a trend of gradual increment was observed up to reactor.
100 P. Tomar, S. Suthar / Desalination 282 (2011) 95103

earthworm activities in reactors. The higher EC of efuent from vermin-
bioltration than experimental control was possibly due to high miner-
alization processes driven by inoculated worm community in reactor
[31]. In general, when organic waste transits through earthworm
gut some fraction of it is then converted into plant available forms
[25]. Probably the release of different mineral ions, such as phosphate,
ammonium and potassium results in increased EC of substrate. The
EC reects the salinity of any material and it is a good indicator of
the mineralize fraction of wastewater. However, after 56 cycles of
treatment EC of efuent water from vermi-bioltration reactor showed
sharp decrement till last observation. This could be due to adsorption
and/or absorption of inorganic constituents of water by different biolog-
ical or non-biological components [32] of vermi-bioltration system.
Moreover, accumulation of salts by inoculated worms, during later l-
tration in vermi-bioltration process, might be responsible for reduc-
tion in EC.
3.3. COD
COD is an important indicator of organic load of urban wastewater.
The COD load in efuents from bioltration and vermi-bioltration sys-
tem was signicantly low than initial levels, but vermi-bioltration
showed more removal efciency than bioltration reactor (t-test:
p b 0.001). The removal rate was 90% (as compared to initial level) in
vermi-bioltration system and 36.8% in bioltartion system. This
could be because earthworms and aerobic microbes act symbiotically
to accelerate and enhance the decomposition of organic matter [33]. Re-
sults clearly indicated the potential of worms in removal of organic load
from wastewater through direct feeding of solid fractions of water or by
promoting microbial-mediated organic decomposition process. The
COD removal rate was 84% in vermi-bioltration system after comple-
tion of 8 cycles of treatment. The changed in COD load of wastewater
during different treatment cycles is illustrated in Fig. 4. In control biol-
tration system COD reduced gradually during treatment cycles while in
vermi-bioltration system COD level of efuent water reduced rapidly
after 1st treatment cycle (Fig. 4). Earlier worker have also reported sig-
nicant reduction in the COD load during bioltration and vermi-
bioltration processes [3, 21]. Sinha et al. [3] studied the vermiltration
of wastewater originated from dairy industry under a pilot-scale project.
They claimed the average COD reduction in the ranges of 8090% at the
end. Under another laboratory trial of urban wastewater treatment
through vermiltration, Sinha and his associates reported about 45% re-
duction in COD load after treatment [19] and removal rate was signi-
cantly high in experimental reactor than control one (without worms).
In general, the geological and microbial system in control bioltration
unit is responsible for COD reduction while in vermi-bioltration system
enzymes, secreted by earthworm and gut-associated microora, reduce
the those chemicals which otherwise cannot be decomposed by mi-
crobes [19, 25]. Recently Xing et al. [21] have reported signicant COD
reduction (47 58% than initial) during vermi-bioltration of domestic
wastewater. Also Ghatnekar et al. [18] have investigated the impact of
vermi-bioltration system on chemical characteristics of wastewater
generated from gelatin industry. They claimed about 90% reduction in
level of COD at the end of process. Wang et al. [5] also reported
90.2% average removal efciencies of vermi-bioltration system for
COD of a domestic wastewater. The microbial association with worms
in vermiltration system could be important for removal of organic
load form wastewater. The presence of earthworm also promotes the
microbial colonization in vermibeds and evidences from recent investi-
gation supports this hypothesis [2, 34]. Zhao et al. [2] investigated the
earthworm-microorganism interaction during wastewater sludge
treatments and results suggested about 46% reduction in the contents
of volatile suspended solids due to earthworm-microbial action after
treatment process. The easy assimilable source of carbon and other
available nutrients from earthworm products, i.e. casts and mucus ac-
celerates the microbial colonization in earthworm-containing vermi-
beds. According to Singleton et al. [35] earthworm hosts millions of
decomposer microbes in their gut and excreate them in soil along
with nutrients in worm casts. Such nutrients further enhance the micro-
bial quality and quality of the vermibeds. Moreover the formation of
biolms of decomposer microbes in the geological system of the ver-
mireactor also promotes COD reduction during vermiltration process
[3]. Chiarawatchai and Nuengjamnong [36] and Chiarawatchai [26] sug-
gested that earthworms contributed to the wastewater remediation
during the treatment process within the VSFCWs. Results thus, clearly
indicate that vermi-bioltration may be an efcient treatment tool for
designing of a low-cost domestic wastewater treatment facility.
3.4. NO3N
Nitrate is an important indicator of water pollution and its high
concentration in freshwater bodies leads to eutrophication problem.
In this study there was signicant impact on nitrate concentration
in efuents after treatment in both experimental reactors. But NO3
reduction arte was prominent in vermi-bioltration unit than biol-
tration system (Table 3). The level of NO3 in efuent after nal treat-
ment cycle was 27.9 mg/L for vermi-bioltration unit and 186.9 mg/L
for bioltration unit. There was about 92.7% total removal of NO3N
in vermi-bioltration unit that was signicantly higher than total re-
moval in bioltration unit (51.3% removal) (t-test: p b 0.001). In tradi-
tional wetland bioltration system the nutrients and metals may be
removed from the polluted water and retained in the sediment and
taken up by the plants and by microorganisms associated on the sur-
face of the roots and sediments, by immobilization in sediments via.,
mechanism such as adsorption on ion exchange sites binding to or-
ganic matter, incorporation into lattice structure and precipitates
into insoluble compounds [37]. The results of removal efcacy of bio-
ltration system were similar to those observed in constructed wet-
lands by other researchers [28, 32] and were moderately higher
than those detected in horizontal ow constructed wetlands planted

Table 3
Chemical characteristics of outlet from bioltration (control reactor) and vermi-bioltration
(experimental) at the end of process.

Parameters Control Experimental t-test c Signicance

reactor a reactor b (t-coefcient value) level

pH 7.81 0.01 8.15 0.01 51.50 P b 0.001

EC (S/cm) 1230.0 8.0 984.67 11.7 22.86 P = 0.002
TSS (mg/l) 78.40 1.00 24.8 2.82 1179.71 P b 0.001
TDS (mg/l) 9875.0 15.0 91.3 5.13 26.113 P b 0.001
NO3 (mg/l) 186.90 1.02 27.92 2.93 69.55 P b 0.001
PO43 (mg/l) 12.67 0.03 0.62 0.04 374.86 P b 0.001
COD (mg/l) 545.6 5.0 86.67 2.03 160.93 P b 0.001
a
Reactor without earthworms (vermi-bioltration).
Fig. 4. Changes in COD during different cycles of treatment in control and experimental b
Reactor with earthworms (bioltration).
reactor. c
Paired sample t-test between control and experiment.
 P. Tomar, S. Suthar / Desalination 282 (2011) 95103
with Phragmites by Vymazal [38] and with Canna and Heliconia by
Konnerup et al. [39]. The high NO3N removal efciency of current
bioltration system than previous reports could be explained in
terms of oxygen supply in rhizosphere of bioltration system. In gen-
eral, oxygen is released by roots of plants in constructed wetlands and
it should be sufcient to meet the demand for the aerobic bacterial
communities which are mainly responsible for NO3N removal
from wastewater. Therefore, addition of aeration device in current
bioltration system was an advantage over traditional bioltration
or wetland ltration systems. Vymazal [38] also pointed out that in
most system designed for the treatment of domestic or municipal
sewage the supply of dissolved organic matter is sufcient and aero-
bic degradation is limited by oxygen availability. The supply of oxy-
gen also promotes the activities of heterotrophic and ammonifying
bacteria which are mainly responsible for nitrate removal from
wastewater. In general, denitrication processes involved the initial
NO3N reduction to NO2N, followed by further reduction to nitric
oxide (NO), nitrous oxide (N2O) and nally to molecular nitrogen
(N2) [40]. However, NO3N reduction rate was relatively high in
this study than previous reports and probably that attributed to oxy-
gen supply in the system. Wang et al. [5] reported efcient removal of
NH4+N (with 85.797.1% of removal rate) while studying wastewa-
ter treatment using an earthworm-based ecological lter integrated
constructed rapid inltration (Eco-CRI) system. Xing et al. [21]
reported about 7.63 to 14.9% total N and 21.0 to 62.3% NH4N removal
in wastewater after treating through vermi-bioltration system. The
different between bioltration system and vermiltration system for
removal rate should be explained in terms of the population and activ-
ities of nitrogen metabolizing bacteria. Chiarawatchai [26] reported sig-
nicant reduction in level of nitrate in efuents obtained from a lab-
scale microcosm wastewater treatment unit than efuent from reactor
without worms. The presence of earthworm in rhizosphere sub-system
has some advantages over traditional bioltration system because of
the direct impact of earthworms on aerobic heterotrophic bacterial
communities which are mainly responsible for N-mineralization in
wastewater bioltration systems. The trend of changing NO3 -N level
during the treatment cycles is described in Fig. 5. There was rapid re-
moval in vermi-bioltration unit than the bioltration unit in the rst
and second cycle of the treatments and that could be due to ltration
of suspended substances during rst cycle of treatment which are con-
sidered to be feed materials for earthworms in vermi-bioltration sys-
tem. Moreover, earthworm-mediated rapid nitrogen transformation
leads to rapid NO3 -N loss from wastewater.
3.5. Phosphate (PO43)
As described in Table 3, there was signicant different between
inlet and outlet water for PO43 concentration in both treatment reac-
tors, i.e. bioltration and vermi-bioltration. The source of phosphate
Fig. 5. Changes in NO3 during different cycles of treatment in control and experimental
reactor.
101
in wastewater is household drains and urban runoff water containing
excreta and other organic substances [21]. The nal efuent from vermi-
bioltration system showed low concentration of PO43 (0.62 mg/L) than
sample collected from nal stage of bioltration system (12.67 mg/L).
The PO43 removal efciency of vermi-bioltration was recorded 98.3%
and that was signicantly higher than removal efciency of bioltration
system (65.2%) (t-test: pb 0.001). The patterns of PO43 removal during
different treatment cycles is described in Fig. 6. It is clear that in bioltra-
tion system the removal trend for PO43 is slow and linear but in vermi-
bioltration reactor there was a trend of sharp PO43 removal up to
56 cycles of treatments thereafter; removal rates declined sharply.
The ligand exchange reactions and physical adsorption or sorption sites
rapidly removes phosphorous from the soil solution. In soil column the
hydroxides and oxides of Al and Fe, calcium carbonate and layer silicate
minerals are important sites for sorption of phosphate anions [41, 42].
The PO43 removal efciency of current bioltration system with Cyprus
stand was relatively higher than those detected in horizontal ow con-
structed wetlands planted with Phragmites by Vymazal [38]. In current
vermitration system the top layer composed of sandy soils along
with mixtures of large stones and pebbles. Probably the sand mixed col-
umn of current bioltration reactor was advantage over the previous
bioltration systems. Preetha and Kumar [43] demonstrated more
than 99% removal of PO43 from wastewater using sand-column treat-
ments device. According to Bostrom et al. [44] aerobic conditions are
more favourable for P sorption and co-precipitation therefore; it is sug-
gested that high PO43 removal could be due to addition of aeration de-
vice in our vermi-bioltration system. However, results of phosphorus
removal contrasts with nding of Chiarawatchai [26] who reported
least impact of earthworm inoculation on phosphorous removal from
wastewater during vermi-bioltration process. He suggested some
technical improvements like replacement of substrates from gravel or
sand to ones with high phosphorus adsorption capacities to enhance
phosphorous removal capability of vermi-bioltration unit. However,
better results of this study than previous report could be attributed to
substrate quality, design and biological components (earthworm spe-
cies, plant type etc.). Moreover, activities of earthworm and associated
microora in vermibeds also promote rapid P-mineralization in the sys-
tem. The level of PO43 in treated efuent from bioltration and vermi-
bioltration is of prime concern because high concentration of such
substance is responsible for eutrophication in surface freshwater re-
sources. Although, the level of PO43 in nal efuent from bioltration
system was comparatively high than prescribed limit, i.e. 5.0 mg/L as
decided by national pollution monitoring agency, i.e. Central Pollution
Control Board (CPCB) for surface discharges of treated water.
3.6. TSS and TDS
Total suspended solids (TSS) and total dissolved solids (TDS)
showed drastic reduction during bioltration and vermi-bioltration
Fig. 6. Changes in PO43 during different cycles of treatment in control and experimen-
tal reactor.
102 P. Tomar, S. Suthar / Desalination 282 (2011) 95103

Fig. 7. Changes in TSS during different cycles of treatment in control and experimental Fig. 9. Changes in different parameters of wastewater during different cycles of treat-
reactor. ment in control and experimental reactor.

process (Fig. 7 and 8, respectively). The total reduction in TDS content domestic wastewater treatment. According to this study earthworm
was about 99.8% in vermi-bioltration unit and that was signicantly presence in treatment system caused about 57 to 79% reduction in
higher than total removal in bioltration system, i.e. 82.6% (t-test: total content of suspended solids in wastewater. The results of present
p b 0.001). Results thus clearly suggested the capability of earthworms study corroborates with the ndings of other scientists who claimed
to remove solid fractions of wastewater during vermi-bioltration importance of earthworm in vermiltration system. However, more de-
processes. Similarly TSS also reduced signicantly in wastewater tailed is needed to establish the relationship between removal of solids
obtained from both experimental reactors (bioltration and vermi- and earthworm working mechanism in vermi-bioltration system.
bioltration) at the end of process. The removal rate was high in
vermi-bioltration unit (88.6% than initial level) than bioltration
4. Conclusions
system (63.8%) (t-test: p b 0.001) (Table 3). The removal pattern of
TDS in both ltration systems is described in Fig. 7. The control (bio-
This work provides an opportunity to explore the efciency of a
ltration) system showed a gradual removal of TDS during different
vermi-bioltration system (mainly constructed by using a wet-
cycles of treatments process while in vermi-bioltartion system TDS
land weed Cyprus rotundus and live biomass of a local earthworm
removed sharply during initial 34 cycles thereafter; the removal
P. sansibaricus) in treatment of urban wastewater. Earlier scientic
process was more or less steady till last observation. The difference
approaches were based upon the use of either plant or earthworm in
between both systems could be due to difference in biological compo-
bioltration unit design. But in this study the integration of these two
nents and working capabilities of both reactors. According to Cooper
components (traditional constructed wetlands system and earthworms
et al. [45] and Vymazal et al. [46] the suspended solids that are not re-
has been applied to design a cost effective and sustainable option for
moved in pre-treatment system are effectively removed by ltration
onsite wastewater treatment. Results clearly suggested that integrated
and settlement processes. Mustafa et al. [47] reported the potential
vermi-bioltration reactor was more efcient than traditional bioltra-
of integrated constructed wetland system with Typha latifolia, Carex
tion system in terms of removal of key chemical pollutant from waste-
riparia, Glyceria maxima, Philarius arundiraecae and Juncus effuses in
water (Fig. 9). Although, results clearly indicates the efcacy of vermi-
wastewater treatments. They have reported signicant removal of
bioltration system in wastewater treatment but further detailed
TSS, i.e. 93.7% after treatment process. According to a study conducted
studies are still required to answer few key issues of this system,
by Prabu and Udayasoorian [28] Phragmitis australis, Cyperus pangorei
e.g. hydrolic load, retention time impact, microbial ecology in ver-
and Typha latifolia planted bioltration system removed about 77, 72
mibeds, earthworm-microbial interaction etc.
and 67%, respectively TSS from wastewater after treatments. The efca-
cy of vermi-bioltration system in TDS and TSS removal is also reported
by earlier authors. Sinha et al. [3] reported total removal of TSS and TDS Acknowledgement
in the ranges of 9092% and 9095%, respectively. They have attributed
the TSS removal to continuous consumption by earthworms. Xing et al. We would like to thank four anonymous reviewers for critical com-
[21] demonstrated the results of a small-scale vermiltration unit for ments and fruitful suggestions on earlier version of the manuscript. The
kind cooperation of laboratory staffs (Mr. Ganesh Bahuguna, Namita
Tiwari, Kamal and Digpal Negi) during experimentation is also acknowl-
edged here.

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