Accepted Manuscript

Vermicomposting of distillery sludge waste with tea leaf residues

Moorthi Mahaly, Abbiramy K. Senthilkumar, Senthilkumar Arumugam, Chitrapriya

Kaliyaperumal, Nagarajan Karupannan

PII: S2468-2039(17)30203-0
DOI: 10.1016/j.serj.2018.02.002
Reference: SERJ 120

To appear in: Sustainable Environment Research

Received Date: 20 July 2017

Revised Date: 27 October 2017
Accepted Date: 7 February 2018

Please cite this article as: Mahaly M, Senthilkumar AK, Arumugam S, Kaliyaperumal C, Karupannan N,
Vermicomposting of distillery sludge waste with tea leaf residues, Sustainable Environment Research
(2018), doi: 10.1016/j.serj.2018.02.002.

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to
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 ACCEPTED MANUSCRIPT

Received 20 July 2017

Received in revised form 27 October 2017

Accepted 7 February 2018

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Vermicomposting of distillery sludge waste with tea leaf residues

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Moorthi Mahalya,*, Abbiramy K. Senthilkumara, Senthilkumar Arumugamb, Chitrapriya

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Kaliyaperumala, Nagarajan Karupannanc
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a
PG and Research Department of Zoology and Wildlife Biology, A.V.C. College (Autonomous),
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Mannampandal 609305, India

b
PG and Research Department of Zoology, Chikkaiah Naicker College, Erode 638 004, India
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c
PG and Research Department of Zoology, Sri Vasavi College, Erode 638 316, India
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* Corresponding author:

E-mail address: moovim24@gmail.com

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ABSTRACT

Studies on bioconversion of solid wastes have been performed for controlling pollution

due to waste disposal. Vermicomposting has been identified as an efficient technology for this

purpose. The present study aims at the management of distillery sludge waste (DSW) combined

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with tea leaf residues (TLR) using the eco-biological tool, vermicomposting. DSW and TLR

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were combined in different ratios and subject to vermicomposting for a period of 45 days using

the earthworm, Eisenia fetida. The physico-chemical properties were analyzed in vermicompost

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samples taken on the 0, 15, 30 and 45th day. It was observed that the values of pH, total nitrogen,

total phosphorus, total potassium, available nitrogen, available phosphorus, available potassium,

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total calcium and total magnesium were increased with a declining trend in electrical
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conductivity, total organic carbon and C/N ratio from its initial value. Finally, the solid wastes

were successfully transformed into a nutrient-rich vermicompost.

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Keywords: Distillery sludge waste, Tea leaf residue, Eisenia fetida, Vermicomposting, Physic-
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chemical parameters

1. Introduction
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In a developing country like India, distillery industries have become a major source of

pollution, as 88% of its raw materials are converted into waste and discharged into the water
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bodies causing water pollution. In India, there are about 579 sugar mills and 295 distilleries.
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These industries generate a large amount of Distillery Sludge Waste (DSW) which is the major

source of aquatic and environmental pollution [1]. Similarly, tea is the most popular beverage

and about 3.5 Mt of tea is consumed annually in the world. A large volume of spent tea leaf and

tea leaf residue is disposed into the environment through daily tea drinking and extraction of

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instant tea and ready-to-drink teas. Many investigations have been conducted to test tea waste for

the field of bioremediation to remove metals [2,3]. Tea and vegetable are important food crops of

the world which are cultivated over an area of more than seven million ha and their annual

production is more than eighty Mt.

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India is the second largest producer of tea; subsequently, the utilization rate is also high.

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Tea leaf residue (TLR) was produced as waste after utilization as a beverage and is generally

disposed off in landfills. Though raw green tea leaf contains tannin, the tannin content is found

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negligible in TLR. This is due to conversion of tannin to tannic acid and is extracted in the tea.

But the lignin is present in a high amount which may enter the water body by leaching from TLR

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during the rainy season. Thus a renowned method of recycling is in need for recycling these
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wastes. As the tea factory products contain a high amount of lignin and low amount of organic

carbon [4], it is best to combine the waste with other carbonaceous waste for recycling. Thus the
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TLR was combined with carbonaceous waste of DSW which can compensate organic carbon
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content. This project was thus intended to recycle both the wastes (DSW and TLR) by
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vermicomposting.

Vermicomposting is getting enormous importance in the amelioration of severe problems

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associated with the disposal of large quantities of organic wastes. It is the most suitable

bioconversion method for solid waste disposal and recycling of organic waste [5]. Many studies
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relating DSW, combining it with cow dung, municipal solid waste and agricultural waste [6-8]
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have been studied. But the efficiency of composting DSW with TLR was not studied yet. Hence,

the aim of the present study was to recycle the DSW and TLR using the earthworm Eisenia

fetida. In short, the conversion of these solid wastes into vermicompost provides a sound solution

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for their disposal as well as the problem of pollution. Also vermicomposting is a low-cost

technology used for conversion of organic waste into organic fertilizers.

2. Materials and methods

In order to achieve the objectives, vermicomposting was done with the help of an exotic

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earthworm species, E. fetida in the Zoology lab, Sri Vasavi College, Erode affiliated to

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Bharathiar University, Coimbatore. The details of the experiment, materials used and analytical

methods are presented below.

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2.1. Collection of DSW and TLR (spent tea powder)

The distillery sludge was collected from a distillery industry located in Erode District,

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Tamil Nadu, India. The TLR was collected from a tea shop close to the study area Sri Vasavi
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College. Both the raw materials were shade dried and physic-chemically analyzed before

implementation.
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2.2. Partial degradation of DSW and TLR

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Though any non-toxic organic wastes can be served as food for earthworms, they cannot
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be directly implemented for vermicomposting. Thus it must be subjected to partial

decomposition and then used as a medium for vermibed. For the process of partial degradation,
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the selected wastes were mixed in different ratios and subjected to degradation by conventional

heap method. The combination of DSW and TLR was done in the following way and the final
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quantity of mixture was made up to 20 kg.

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Treatment I - 60% DSW + 40% TLR (12 + 8 kg)

Treatment II - 70% DSW + 30% TLR (14 + 6 kg)

Treatment III - 80% DSW + 20% TLR (16 + 4 kg)

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The DSW and TLR were shade dried, powdered and sieved using 2 mm mesh. Then they

were weighed and arranged in alternate layers as a heap spread on a clear floor of a thatched

shed. Cow dung mixed in portable water was used as inoculum and was sprinkled in between

each layer. 50% of moisture was maintained in the heap and mulching was done with thatched

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coconut leaves. Partial degradation was carried out for 40 d. The wastes were turned over with a

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spade once in a week. The microorganisms present in the cow dung start degrading the solid

wastes. After 40 d the partially decomposed DSW and TLR were recovered and used as a

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medium for vermibed.

2.3. Vermicomposting

2.3.1. Preparation of vermibeds

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Rectangular plastic containers (bins) of 45 cm height, 40 cm length, and 25 cm width

were chosen for vermicomposting. Vermibeds were prepared with the partially degraded DSW
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and TLR (various ratios). Due to partial degradation, the whole amount (20 kg) of DSW and
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TLR were reduced to approximately three-fourths, or only 15.345 kg was obtained after 40 d
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which were subject to vermicomposting in five replicates. The process was carried out in

Vermicomposting lab of Zoology Department, Sri Vasavi College.

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2.3.2. Inoculation of earthworms

About 25 young non-clitellate earthworms E. fetida, weighing not less than 200-250 mg
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were introduced in the vermibeds. The moisture level of the containers was maintained around
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35-40% throughout the study by sprinkling water. All containers were incubated in a humid and

dark place at room temperature and were kept undisturbed and allowed to produce vermicast.

They were covered by a net to avoid the escape of worms and to safeguard from pests. E. fetida

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(commonly called as compost worm or red worm) is an epigeic worm and hence produces the

castings on the surface of the vermibed.

The vermicast was collected periodically, shadow dried, grounded and sieved. The

physic-chemical parameters, pH, electrical conductivity (EC), total organic carbon (TOC), total

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nitrogen (TN), total phosphorus (TP), total potassium (TK), available nitrogen (AN), available

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phosphorus (AP), available potassium (AK), carbon/nitrogen ratio, total calcium (TCa), and total

magnesium (TMg) were analyzed in the vermicast collected periodically (0, 15, 30 and 45th

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days). I changed to lowercase

2.4. Analytical methods

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The following standard methods were followed to analyze the vermicompost samples.
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The pH and EC were analyzed through pH meter and conductivity meter in sample water

suspension at 1:10 ratio [9]. TOC was analyzed by chromic acid wet digestion method [10] and
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Macro Kjeldahl method for the analysis of TN followed by Humphries [11]. TP was analyzed by
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colorimetric method, molybdenum in sulphuric acid by Van ado molybdate yellow color method
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and TK was analyzed by flame photometric method [12]. AN, AP and AK were analyzed by

following Subbiah and Asija, Olsen et al. and Black method respectively [13-15]. TCa and TMg
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were analyzed by triple acid digestion and extraction (HNO3:H2SO4:HCIO4 at 9:2:1) - Versenate

method [16].
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2.5. Statistical analysis

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Two-way analysis of variance was computed using SPSS (version 22) to test the level of

significance of the difference between various vermicompost with respect to the physic-chemical

parameters. T-test analysis was conducted to calculate the p-value.

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3. Results

3.1. Physic-chemical parameters of raw materials

The initial physic-chemical parameters of DSW, cow dung (CD) and TLR were analyzed

and presented in Table 1. The main parameters to be considered were the TOC of DSW and the

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lignin and tannin content of the TLR. The value of TOC was very high in DSW and was creating

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an odor problem in the environment. When the lignin and tannin content were analyzed in TLR,

lignin was comparably high than the other two (CD and DSW; Table 1) which may leach out and

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pollute the water bodies during the rainy season. Tannin content was non-detectable in all the

three raw materials. Usually, tannin is the major pollutant present in the tea leaf. But as the tea

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leaf is boiled it gets extracted in the tea extract and a negligible amount is seen in the TLR.
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Tannin, when diluted with water, turns into tannic acid and dissolves in the extract [4]. Thus the

raw materials combined in different ratios were partially degraded using CD as inoculum.
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3.2. Vermicomposting
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The casting activity of E. fetida was found to be increased with the increase in TLR
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content i.e. in the order, treatment I > treatment II > treatment III. The highest casting activity

was found in unit of 60% DSW. This infers the fact that earthworms preferred less DSW with
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more TLR content. This may be due to the C/N ratio which is found favorable in the 60:40 ratio

than others. The presence of high amount of TOC leads to an unpleasant odor. Hence the
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assimilation process was seen more in 60:40 ratio. The 80% DSW with 20% TLR showed a
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lower rate of assimilation and casting activity. Thus their physic-chemical characters were

reduced limitedly when compared to other two in the 45th day (Tables 2 to 5).

3.3. Physic-chemical parameters of vermicomposted DSW and TLR

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The Physic-chemical values of the vermicomposted DSW and TLR in different

combinations were presented in Tables 2 to 5. The characterization was done in different time

intervals (i.e., initial day, 15, 30 and 45th day). The 0th day parameters represent the physic-

chemical parameters of the partially degraded DSW and TLR combinations. Table 2 shows the

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values of pH, EC, and TOC. The results were almost similar in all the three different

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combinations. The pH was acidic in nature during the initial stage while it was nearing neutral

during 15th day. On 30 and 45th days, the pH subsequently reached neutral in all the three

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treatments. EC was gradually declining from 0 to 45th day (Table 2). Nearly 50% of EC was

reduced in the 45th day castings. This neutral pH and reduced EC of vermicompost enables the

plant growth which when applied as manure.

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As the process of vermicomposting proceeds, the TOC level decreased. The TOC ranged

from 42 to 45% in the different ratio of DSW with TLR combinations on the initial day.
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Naturally, the DSW is highly rich in TOC. Thus the ratios with high percentage of DSW
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contained high level of TOC. The TOC range was abridged on all the 45th day vermicomposts
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when compare to the initial stage (Table 2). This significantly decreased level of TOC might be

the reason for the reduced bad odor of DSW. Though the lignin content measured in raw TLR
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was high, it was decreased to non-detectable level after partial degradation and hence was not

measured in the following consecutive days.

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The major nutrients, i.e., the total and AN, TP and TK content analyzed in DSW and
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TLR combinations are presented in Tables 3 and 4. Higher concentration of nutrients was

observed at end of vermicomposting in all the three ratios. The increase was gradual as the days

goes on (Fig. 1). About 28% increase in TN% was recorded in 60, 70 and 80% DSW

combination. AN was observed assortment in 0.38 to 0.55% in 60:40 ratio; 0.24 to 0.42 in 70:30

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and 0.19 to 0.37% in 80:20 ratio of DSW with TLR combinations (Table 4). Thus the increase in

nitrogen level was found invariable and independent of the ratios. In the whole, the important

plant nutrients, NPK were found increasing in the vermicomposted DSW and TLR.

The other micronutrients and C/N ratio analyzed are presented in Table 5. TCa and TMg

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levels were increased with time. The 60% ratio of DSW already contained a higher amount of

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TCa, thus the amount in the final day was also found to be increased when compared to the other

two (Table 5). Thus the vermicomposted DSW and TLR is also rich in the micronutrients which

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infer the fact that a superior quality vermicompost can be prepared from the solid wastes DSW

and TLR.

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Consequently, after vermicomposting changes in C/N ratios were significant in the 45th
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day from its initial stage and those present in the raw materials (16; 21 and 25). This indicates

that the undergoing decomposition was more intense and an apt C/N ratio (16 to 24) was
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obtained in all the three combinations of DSW and TLR (Table 4). The C/N ratios are far much
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reduced from its original state and the solid wastes have been recycled into a perfect
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vermicompost. Among the three ratios experimented, 60: 40 ratio was found to be suitable for

vermicomposting.
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4. Discussion

The vermicompost was much darker in color and had been processed as more or less
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homogenous mixture after 45 days of earthworm’s activity. The vermicomposting significantly

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modified the physic-chemical properties of different carbonaceous mixtures tested. The pH of all

the vermicompost was almost neutral and stabilized at matured stage (45 days) which may be

due to the buffering nature of humic substances [17]. The similar results were observed by

Nagavallemma et al. [18]. Similar studies made on vermicomposting of DSW with municipality

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waste showed that the decomposition process removed the unpleasant odor and increased the

number of earthworms considerably [19].

In all ratio of vermicompost, a significant decline in EC from its initial day was observed.

The declination of EC might be due to the utilization of soluble salts by microorganisms for the

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synthesis of microbial biomass [20] and also due to the absorption of soluble salts by earthworms

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and enhanced microbial activities [21]. Significantly lower TOC of all the vermicomposts

suggested enhanced oxidation of organic carbon, as the results of the increased microbial

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activity. The loss of TOC may be due to the conversion of CO2 through microbial respiration and

mineralization of organic matter causing an increase in total nitrogen. The microorganisms use

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the carbon as a source of energy decomposing the organic matter.
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C/N ratio is one of the most widely used indicators of vermicompost maturation,

decreased sharply during vermicomposting. Generally, the C/N ratio of vermicomposting ranges
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from 14 to 22 [22]. The C/N ratio in the present results is within this range except in 80:20 ratio
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of distillery sludge with tea leaf residue combination. This may be due to the high amount of
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carbon present in the distillery sludge. According to Senesi [23] declined C/N ratio (less than 20)

is the indications of the advanced degree of organic matter stabilization and reflects a satisfactory
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degree of organic wastes.

Significantly higher TN content suggested the superior composting ability of E. fetida.

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Vermicompost contains a comparatively high level of nitrogen compared to other bulky organic
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manures. Generally, TN content in the vermicomposting range from 1.5 to 2.7% [24]. The

increase in TN may be due to the mineralization of the organics present in DSW and TLR.

Similar results were also shown by Carbera et al. [25] in their work. The other nutrients, TP, TK,

AP and AK level in vermicompost indicate that a higher mineralization rate occurs during the

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process of vermicomposting. Earthworms turn the insoluble and particulate organic matter into

soluble forms with the help of solubilizing microorganisms present in the gut making it more

available for the future plantations [26]. Significantly higher amount of nutrients in

vermicomposts might be due to more effective humification in these treatments [27].

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When the micronutrients level was monitored, higher TCa and TMg content was

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recorded in the matured vermicomposts produced by E. fetida. Higher concentration of

micronutrients in vermicompost was also reported in earlier workers [28,29]. Thus the solid

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wastes, DSW along with TLR was recycled into a nutrient-rich vermicompost successfully in

this project with the help of the earthworm, E. fetida.

5. Conclusions
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Vermicomposting was adopted for the recycling of solid wastes, DSW and TLR in

different combinations involving the earthworm, E. fetida. Periodical physic-chemical analysis

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(0, 15, 30 and 45th day) of different combinations of vermicompost was studied. pH, Total N, P
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and K, Available N, P and K, TCa and TMg were observed in increasing trend while EC, TOC
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and C/N ratio were observed in declining trend from its initial days of vermicomposting.

Considering the above facts, the present study concludes that vermicomposting of DSW and
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TLR is possible with the bioconversion of these organic wastes into manure probably

minimizing the environmental pollution caused by their disposal.

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Table 1. Physic-chemical values of raw DSW, CD and TLR

Physic-chemical values DSW CD TLR

pH 5.47 7.62 5.71
EC (dS m-1) 0.71 ± 0.09 3.80 ± 1.53 0.12 ± 0.03

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TOC (%) 62.4 ± 2.2 34.3 ± 1.9 3.9 ± 0.5
TN (%) 0.12 ± 0.03 2.30 ± 0.14 0.09 ± 0.01
TP (%) 5.63 ± 0.24 0.42 ± 0.06 1.62 ± 0.03

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TK (%) 0.98 ± 0.02 0.49 ± 0.03 0.81 ± 0.06
AN (%) 0.12 ± 0.01 1.15 ± 0.06 0.09 ± 0.03
AP (%) 5.0 ± 1.0 3.7 ± 1.0 1.1 ± 0.3

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AK (%) 0.08 ± 0.03 0.12 ± 0.08 4.03 ± 1.06
C/N Ratio 520 15 43
TCa (%) 0.62 ± 0.08 0.47 ± 0.02 0.21 ± 0.07
TMg (%) 0.41 ± 0.05 0.39 ± 0.23 0.38 ± 0.08

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Lignin (%) 0.03 ± 0.09 1.32 ± 0.91 16.34 ± 2.45
Tannin (%) ND ND ND
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P-Value 0.001* 0.001* 0.001*
ND: Non-detectable * Significant (P < 0.05)
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1 Table 2. pH, electrical conductivity and total organic carbon in vermicompost prepared from different combinations of DSW and
2 TLR.
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pH EC (dS m-1) TOC (%)
Days

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60:40 70:30 80:20 60:40 70:30 80:20 60:40 70:30 80:20
0 6.29 6.54 6.32 0.66 ± 0.11 0.67 ± 0.06 0.67 ± 0.12 42 ± 1 44 ± 1 45 ± 0.5

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15 6.82 6.87 6.81 0.54 ± 0.14 0.56 ± 0.13 0.56 ± 0.11 40 ± 1 42 ± 2 44 ± 1
30 7.04 7.01 7.05 0.42 ± 0.09 0.41 ± 0.09 0.40 ± 0.13 39 ± 1 39 ± 1 41 ± 1

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45 7.09 7.12 7.01 0.39 ± 0.06 0.39 ± 0.06 0.39 ± 0.11 35 ± 0.5 37 ± 1 39 ± 0.5

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P-Value 0.001* 0.001* 0.0001*
4 * Significant (P < 0.05)

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13 Table 3. Total nitrogen, total phosphorus and total potassium of vermicomposted DSW and TLR in different combinations.

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TN (%) TP (%) TK (%)

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Days
60:40 70:30 80:20 60:40 70:30 80:20 60:40 70:30 80:20
1.5 ± 0.2 1.3 ± 0.1 1.1 ± 0.2 2.7 ± 0.0 3.5 ± 0.0 4.4 ± 0.1 0.7 ± 0.1 0.8 ± 0.1 1.0 ± 0.2

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0
1.6 ± 0.1 1.5 ± 0.1 1.3 ± 0.1 2.8 ± 0.0 3.6 ± 0.0 4.5 ± 0.0 0.9 ± 0.0 1.0 ± 0.0 1.2 ± 0.0

AN
15
30 1.9 ± 0.1 1.6 ± 0.2 1.4 ± 0.1 2.9 ± 0.0 3.7 ± 0.0 4.8 ± 0.0 1.0 ± 0.0 1.2 ± 0.1 1.5 ± 0.1
2.1 ± 0.1 1.8 ± 0.0 1.6 ± 0.0 2.9 ± 0.0 3.8 ± 0.0 4.8 ± 0.0 1.2 ± 0.1 1.4 ± 0.1 1.5 ± 0.1

M
45
P-Value 0.0005* 0.0006* 0.001*

D
14 * Significant (P < 0.05).

TE
15
EP
16
C

17
AC

18

19

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20

21 Table 4. Available nitrogen, available phosphorus and available potassium in vermicompost prepared from different combinations of

PT
22 DSW and TLR.

RI
AN (%) AP (%) AK (%)

SC
Days
60:40 70:30 80:20 60:40 70:30 80:20 60:40 70:30 80:20

0.37 ± 0.10 0.24 ± 0.03 0.19 ± 0.12 2.14 ± 0.02 3.11 ± 0.09 4.09 ± 0.01 0.14 ± 0.11 0.13 ± 0.42 0.12 ± 0.13

U
0

AN
15 0.42 ± 0.09 0.31 ± 0.06 0.23 ± 0.10 2.16 ± 0.01 3.13 ± 0.01 4.11 ± 0.01 0.18 ± 0.52 0.17 ± 0.90 0.21 ± 0.81

30 0.46 ± 0.10 0.36 ± 0.12 0.31 ± 0.10 2.17 ± 0.06 3.15 ± 0.04 4.14 ± 0.01 0.22 ± 0.51 0.23 ± 0.41 0.26 ± 0.11

M
45 0.55 ± 0.01 0.41 ± 0.01 0.36 ± 0.08 2.19 ± 0.01 3.17 ± 0.09 4.15 ± 0.03 0.29 ± 0.63 0.28 ± 0.82 0.30 ± 0.92
P-Value 0.0005* 0.0001* 0.0001*

D
23 * Significant (P < 0.05)

24
TE
EP
25
C
AC

26

27

28

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30 Table 5. Carbon/nitrogen, total calcium and total magnesium in vermicompost prepared from different combinations of DSW and

PT
31 TLR.

RI
C/N ratio Ca (%) Mg (%)
Days

SC
60:40 70:30 80:20 60:40 70:30 80:20 60:40 70:30 80:20
0 28 ± 0 34 ± 1 44 ± 1 1.0 ± 0.1 0.9 ± 0.0 0.8 ± 0.0 0.8 ± 0.3 0.6 ± 0.2 0.5 ± 0.1

U
15 40 ± 1 42 ± 2 44 ± 0 1.4 ± 0.1 1.2 ± 0.1 1.0 ± 0.1 1.2 ± 0.1 1.1 ± 0.1 1.0 ± 0.1

AN
30 21 ± 1 24 ± 1 30 ± 1 1.9 ± 0.1 1.7 ± 0.1 1.3 ± 0.1 1.3 ± 0.1 1.1 ± 0.2 1.0 ± 0.2

17 ± 1 21 ± 1 25± 1 2.0 ± 0.1 1.9 ± 0.1 1.6 ± 0.1 1.4 ± 0.2 1.4 ± 0.0 1.3 ± 0.0

M
45
P-Value 0.0005* 0.049* 0.0001*

D
32 * Significant (P < 0.05)

33
TE
EP
34
C
AC

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PT
RI
U SC
AN
35

M
36 Fig. 1. Chart showing the Total N, P and K analyzed in 60: 40 combination in different periods.

D
TE
C EP
AC