Bioresource Technology 293 (2019) 122047

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

Fly ash as an additive for enhancing microbial and enzymatic activities in in- T
vessel composting of organic wastes
⁎
Ashootosh Mandpea,b, Sonam Paliyaa,b, Sunil Kumarb, , Rakesh Kumarb
a
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 2010 02, India
b
CSIR-National Environmental Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur 4400 20, India

G R A P H I C A L A B S T R A C T

A R T I C LE I N FO A B S T R A C T

Keywords: In the present study, fly ash was used as an additive for the rapid composting of biomass and kitchen waste with
Microbial jaggery as an activator. The in-vessel composting experiments were carried out in ten plastic vessels of 15 L
Compost capacity containing varying amounts of fly ash, biomass waste, and kitchen waste. The effect of fly ash on the
Dehydrogenase activity microbial and enzymatic activity of the composting system was also ascertained. The compost in Vessel No. 10
Fly ash
having a combination of 50% biomass waste, 30% kitchen waste, and 20% fly ash seemed to have a significant
Substrate
effect on the compost quality. Statistical analysis revealed Vessel No. 10 as significantly separated from other
vessels whereas microbial activity (342.67 CFU 108/g), pH (8.15), temperature (48.50 °C), and heavy metals,
such as Ni (0.1239 mg kg−1), Ba (0.6109 mg kg−1), Cd (0.0004 mg kg−1) were the most influential variables for
the separation.

1. Introduction rate of waste generation is in direct proportion to the present popula-

It is generally observed that economic prosperity and larger the
number of the urban population led to a higher generation rate of solid
wastes. According to the United Nations Department of Economic and
Social Affairs (UNDESA) 2017, it is projected that the total world po-
pulation may reach 8.6 billion by the year 2030. It was found that the
tion; however, the waste characteristics may vary depending on dif-
ferent factors including income groups, food habits and standard of
living of the masses (Population Reference Bureau, 2017). In the Indian
context, solid waste management (SWM) was continued to be the most
neglected part of the infrastructural development. The infrastructural &
economic constraints, coupled with space unavailability for discarding

⁎
Corresponding author.
E-mail address: s_kumar@neeri.res.in (S. Kumar).

https://doi.org/10.1016/j.biortech.2019.122047
Received 15 July 2019; Received in revised form 17 August 2019; Accepted 20 August 2019
Available online 22 August 2019
0960-8524/ © 2019 Published by Elsevier Ltd.
A. Mandpe, et al.
wastes and lack of awareness among the common masses led to re-
straining the progress in the efficient SWM practices (Beliën et al.,
2014).
For fulfilling the food requirements of the ever-increasing popula-
tion, the agrarian production has been intensified rapidly by the ap-
plication of chemical fertilizers and pesticides which suppresses the
quality of soil (Kumar et al., 2009). In India, the crop production with
other allied activities associated with agriculture generate around 500
Million Tonnes (MT) of biodegradable lignocellulosic wastes and dis-
posing this tremendous amount of wastes within a short period is a
major challenge for the researchers and the environmental planners in
India (NPMCR, 2014; Bhuvaneshwari et al., 2019). To manage the
enormous volume of biodegradable wastes effectively, several tech-
nologies like anaerobic digestion, pyrolysis, incineration, and gasifica-
tion can be implemented, however; composting is a favourable option
as it is comparatively easier to operate and manage. The waste stream
can be processed biologically, and the end product is well-stabilised
humus, which can be utilized as a soil conditioner (Adewumi et al.,
2005).
Similarly, to cope up with the surging demand of energy due to the
increased urbanization coupled with rapid industrialization, several
coal-fired thermal power plants have developed all over the world
(Asokan et al., 2005). The combustion of bituminous, sub-bituminous
and lignite coal for energy production in these thermal power plants
generate huge quantities of fly ash (FA), bottom ash, flue gas desul-
phurization (FGD) material and boiler gas as a by-product (Kishor et al.,
2010). Disposal of such behemoth volumes of FA is also one of the
major issues in developing countries like India. In the Indian context,
the primary source for power generation is coal, which accounts for
approximately 56.8 per cent of total power generation capacity (CEA
Report, 2018). The alternate sources of energy are required to be de-
veloped for reducing the stress on excessive utilization of coal for en-
ergy production.
FA, which is being dumped in ash ponds by thermal power stations
occupies around 1,00,000 ha of land in India (Jha and Kumar, 2011)
and disposal of such a tremendous amount of FA has become an en-
vironmental threat since it has been an utterly under-utilized resource
for a long time. There are numerous reasons behind the under-utiliza-
tion of FA, such as lack of awareness, stringent policies regarding FA
management, and the most critical factor is the readily available land. It
has been found that the percentage utilization of the generated FA in
Denmark is 100%, 85% in West Germany, 85% in France, 50% in the
United Kingdom, 45% in China and it is only 38% in India (Kishor et al.,
2010). FA is enriched with both macro & microelements and also
contains heavy metals, such as Cadmium (Cd), Lead (Pb), Nickel (Ni),
Chromium (Cr), Zinc (Zn), Molybdenum (Mo), Arsenic (As) and Copper
(Cu) (Tiwari et al., 2008). Depending on the sulphur contents present in
the parental coal, the pH ranges from 4.5 to 12.0 (Naiya and Das,
2016). FA contains all the elements similar to those present in soil ex-
cept Nitrogen and a negligible amount of organic carbon (Banerjee
et al., 2014). Thus, except Nitrogen, all other essential elements re-
quired for the metabolism and growth of microbes and plants are
readily available in FA.
In today’s scenario, FA is used for different potential applications
including cement production, pavement construction, manufacturing
bricks, synthesis of zeolites, manufacturing of pavement blocks and has
also been used as fillers in the polymers (Abdel Salam et al., 2011). It
has been used as a soil conditioner for the amelioration of soil quality.
Pappu et al. (2007) reported that FA could also be used for the ex-
traction of heavy metals and treating the wastewater. Wong et al.
(2009) suggested that instead of lime powder, FA can be utilized due to
its unique mechanism which can defile the bioavailability of heavy
metals using physical adsorption and precipitating at a higher range of
pH. FA contains MgO and CaO due to which heavy metals get pre-
cipitated in the soil after the addition of FA to it (Wong et al., 2010;
Singh and Kalamdhad, 2013).
2
Bioresource Technology 293 (2019) 122047
Considering utilization of FA in various applications, it can be un-
derstood that despite FA has numerous applications in diverse fields,
the use of FA in agriculture is limited as compared to other sectors as
well as a behemoth volume of FA remains unutilized. Several studies
reported that FA has an elemental property which improves soil health
and crop yield without causing any phytotoxicity on plant composition
(Katiyar et al., 2012) still the applicability of FA in composting process
has rarely been tested (Lau et al., 2001).
Keeping this in view, the present study was carried out to use FA as
an additive in in-vessel composting of organic wastes for its sustainable
utilization and to evaluate its effect on the microbial and enzymatic
activity as well as the quality of compost. The current work is first of its
kind to use FA in rapid composting of biomass and kitchen waste for its
utilization in an environment-friendly manner and to boost its applic-
ability at a commercial level.
2. Materials and methods
2.1. Collection of FA and organic waste
FA samples were collected from Khaperkheda Thermal Power
Station (TPS) located at 21.276138°N 79.121518°E in Nagpur district of
Maharashtra, India. Biomass wastes (BW) comprising agricultural crop
residue was collected from the farmlands located in outskirts of the city,
dry leaves and branches were collected from trees shedding in the CSIR-
NEERI premises. Also, the kitchen waste (KW) was sampled out from
the canteen of CSIR-NEERI campus.
2.2. Physico-chemical analysis of FA, substrate, and compost
The physico-chemical characterization of FA, organic waste sub-
strate and compost samples were performed for analyzing pH using
Mettler Toledo digital probe; moisture content following gravimetric
method; total carbon, total nitrogen, carbon: nitrogen ratio and sulphur
contents using Elementar Vario EL CHNS Analyser. nitrate, sulphate,
phosphate using UV-1650pc, UV-VIS Spectrophotometer SHIMADZU;
and heavy metal analysis using ICP-OES, iCAP 6000 SERIES. Cellulose
and Hemicellulose were analyzed by adopting the method followed by
Liu (2004), and Lignin content was determined using Klasson's method.
2.3. Pre-treatment of substrate
The waste substrates were pre-treated before processing in an in-
vessel composting system. BW was shredded into smaller parts ranging
in 0.5–1-inch in size resulting in increased surface area and porosity,
which also aids in mixing during the process of composting (Rawoteea
et al., 2017). The smaller-sized organic waste material was further
subjected to steam treatment to breakdown the higher lignocellulosic
contents.
2.4. Experimental Set-up
In-vessel composting experiments were carried out in ten plastic
vessels, each having 15 L capacity with a height of 32.5 cm. The top and
the base diameter of the vessels were 28 and 23.6 cm, respectively. The
vessels were drilled in the bottom part for leachate collection, and the
internal assembly involved the use of perforated rubber tube of 5 mm
diameter, and its end was joined to the aerating pump for maintaining
the aeration rate of 4 L per minute (LPM) during the composting pro-
cess.
Each vessel contained a different combination of BW, KW, FA and
jaggery as an additive along with 50 ml of microbial consortia con-
taining Trichoderma viride, T. harzianum, and T. reesei. The different
combinations of waste substrate applied for in-vessel composting are
presented in Table 1. In vessel 1 only BW (1000 g) was loaded, while
150 g jaggery was added as an activator in Vessel 2 along with the
A. Mandpe, et al. Bioresource Technology 293 (2019) 122047

Table 1 pcs = Average TPF concentration of the test sample

Combination of different substrates with FA in different vessels. pbs = Average TPF concentration of the control sample
Sr. No. Vessel Fly Additive Microbial Biomass Kitchen V = Volume of substrate extractant
ash (Jaggery) mixture (ml) waste (BW) waste m = Mass of Soil sample in gram
(FA) (g) (g) (KW) (g) t = Time of incubation in hours
(g)

1. V 1 X X 50 1000 X 2.7. Statistical analyses

2. V 2 X 150 50 1000 X
3. V 3 100 X 50 900 X
All the calculations and the statistical analyses were performed
4. V 4 200 X 50 800 X
5. V 5 300 X 50 700 X using Microsoft Office Excel and Minitab Software (Version 18)
6. V 6 100 150 50 900 X (Chavarria et al., 2018). The data of pysico-chemical properties of FA,
7. V 7 200 100 50 800 X organic waste substrates and the compost samples along with the mi-
8. V 8 300 50 50 700 X crobial population and the enzymatic activity were subjected to ana-
9. V 9 200 X 50 500 300
10. V 10 200 50 50 500 300
lysis of variance (ANOVA), and least significant difference (LSD) at
p ≤ 0.05 was calculated to test the difference between the parameters.
Principal component analysis (PCA) was performed to identify the best
ingredients of Vessel 1. The BW in vessels 3, 4, and 5 was amended with contributing variables for the separation of treatments. Also, the cor-
10%, 20% and 30% FA, respectively on dry basis, while the BW in relation was assessed among the microbial population, heavy metals
vessels 6, 7 and 8 was mixed with 10%, 20% and 30% dry FA along concentration, temperature, and pH using Pearson's coefficient and
with 150 g, 100 g and 50 g of activator, respectively. Vessel 9 was p ≤ 0.05 and p ≤ 0.01 were used to indicate statistical significance.
loaded with 50% BW, 30% KW and 20% dry FA, and in Vessel 10, 50 g
of jaggery was added as an activator along with the ingredients of the 3. Results and discussion
Vessel 9.
3.1. Initial physico-chemical characteristics of FA and substrates
2.5. Estimation of microbial population
The initial physico-chemical characteristics of organic wastes (BW
The microbial population in all the vessels with varying con- and KW) and FA (mean ± standard deviation) are presented in
centrations of additives was determined by serial dilution and spread Table 2. The FA collected for the study was recorded as neutral with the
plate technique. One gram of each combination was weighed and mixed pH value of 7.05 and substrates used for composting were observed
with 9 ml of sterile distilled water in a test tube and then serially diluted slightly acidic, i.e., 5.84 and 6.08 for BW and KW, respectively. The
by 10-fold dilution method up to 10-9 dilution. Further, 0.1 ml of each higher moisture content (MC) was observed in KW (83.50%) followed
sample was inoculated over nutrient agar media petri plates with the by BW (44.96%). The high MC of KW served as a moisture source for
composition of peptone 0.5 g/l along with beef extract 0.3 g/l, sodium microbes during the process of composting. The maximum cellulose,
chloride (NaCl) 0.5 g/l and 1.5 g/l agar. These petri plates were in- hemicellulose, and lignin contents were found in BW followed by KW.
cubated at 37 ± 2 °C for 72 h and post-incubation, the colony forming The cellulose, hemicellulose and lignin contents were not determined
units (CFU) were estimated for viability assessment. for FA as it is the by-product of coal combustion in the thermal power
station. The micronutrients like phosphate (P) and sulphate (S) were
2.6. Measurement of enzymatic activity in vessels found to be in higher proportion in BW as compared to KW and FA.
However, the nitrate content was more in KW as compared to BW. The
Dehydrogenase activity of the total microbial population in all the carbon and nitrogen were found to be in a negligible amount (0.09%
combinations was determined by adopting the protocol followed by and 0.04%, respectively) in FA, whereas in KW and BW, the respective
Klein et al. (1971) with few modifications. One gram of each sample contents were 40.87% and 17.18% for total carbon and 3.03% & 0.96%
was taken in a screw-capped test tube and 1 ml of 0.6% solution of for nitrogen as presented in Table 2. The C:N ratio of BW and KW was
triphenyl tetrazolium chloride (TTC) along with 1 ml of 1% glucose found to be 17.62 and 22.09, respectively, which is recorded in a
solution, 0.02 g calcium carbonate (CaCO3) and 1 ml of sterile distilled negligible amount in FA. According to Wan Razali et al. (2012), the
water was added in all the tubes. Further, the solution was thoroughly nutrients detected in the organic wastes applied for composting were
mixed, and the bottom of each test tube was tapped mildly to prevent found to be suitable, since generally, the values of N and P should be
discharge of trapped oxygen. Later, the test tubes were incubated at
37 ± 2 °C for 24 h. After incubation, the reduced tri-phenyl formazan Table 2
was extracted using 10 ml ethanol by shaking vigorously. Physico-chemical characterization of substrates.
Further, the test tubes were centrifuged for 10 min at 3000 RPM
Sr. No. Parameters Biomass waste Kitchen waste Fly ash (FA)
using the REMI R-8C Plus model. Clear fade pink coloured supernatants (BW) (KW)
were collected and quantified by measuring the absorbance at a wa-
velength of 485 nm by UV-1650PC, UV–VIS Spectrophotometer SHIM- 1. pH 5.84 ± 0.04 6.08 ± 0.05 7.05 ± 0.05
ADZU. The dehydrogenase activity was determined in terms of the 2. MC (%) 44.96 ± 0.48 83.50 ± 0.4 0.017 ± 0.02
3. Cellulose (%) 79.50 ± 0.4 36 ± 0.42 ND
concentration of formazan formed in the samples by comparing it with 4. Hemicellulose (%) 80.50 ± 0.4 32 ± 0.5 ND
graded concentrations of formazan. The results were indicated as mi- 5. Lignin (%) 81.50 ± 0.4 44.5 ± 0.3 ND
crogram (µg) of triphenyl formazan (TPF) formed per gram of sample 6. Nitrate (mg g-1) 16.25 ± 0.24 59.08 ± 0.3 0.01 ± 0.01
per hour (μg TPF h−1 g−1). Dehydrogenase activity was calculated 7. Phosphate (mg g-1) 17.25 ± 0.24 18.26 ± 0.05 5.261 ± 0.001
8. Sulphate (mg g-1) 109.68 ± 0.2 3.04 ± 0.04 5.65 ± 0.07
using Eq. (1).
9. TC (%) 17.18 ± 0.08 40.87 ± 0.11 0.09 ± 0.03
α = [(pcs − pbs)xVx100]/(mxt) (1) 10. TN (%) 0.96 ± 0.07 2.03 ± 0.04 0.04 ± 0.05
11. C:N 17.62 ± 0.04 22.09 ± 0.06 2.25 ± 0.04
where, 12. S (%) 0.09 ± 0.005 18.99 ± 0.06 0.92 ± 0.003

α = Dehydrogenase activity (µg/g/hr) Columns represent mean of three replications and standard error bars; ND: Not
Detected.

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A. Mandpe, et al. Bioresource Technology 293 (2019) 122047

Table 3
Analysis of heavy metal in substrates used for composting.
Sr. No. Heavy metals (mg kg−1) Biomass waste (BW) Kitchen waste (KW) Fly ash (FA) Indian permissible limit (Awasthi and Bhatnagar, 2000)

1. Cd BDL BDL 0.44 ± 0.005 3.6

2. Co 2.08 ± 0.06 0.06 ± 0.008 5.06 ± 0.06 N/A
3. Cr 7.97 ± 0.15 3.49 ± 0.06 18.39 ± 0.06 N/A
4. Cu 15.29 ± 0.06 6.82 ± 0.05 12.49 ± 0.06 135–270
5. Fe 17.29 ± 0.06 140.16 ± 0.10 489.7 ± 0.50 N/A
6. Mn 50.25 ± 0.3 9.07 ± 0.09 86.39 ± 0.39 N/A
7. Ni 4.09 ± 0.06 0.04 ± 0.05 13.97 ± 0.05 75–150
8. Pb 1.80 ± 0.15 0.41 ± 0.02 4.05 ± 0.05 250–500
9. Zn 4.80 ± 0.15 23.04 ± 0.5 5.13 ± 0.3 300–600

Columns represent mean of three replications and standard error bars; BDL-Below Detection Limit.

more than 1% for providing the adequate nutrition to the plants.

The total heavy metal concentration was found to be higher in FA in
comparison to the organic waste substrate as presented in Table 3,
whereas macronutrients, such as N and C were observed more in the
organic waste substrates. From Table 3, it can be perceived that the
concentration of heavy metals present in FA is less than the permissible
limit, and it was also added in a smaller proportion in the composting
system. Thus it provides essential nutrients, such as P, Mn, Fe, and Cu
for the microbial growth in compost which will further lead to the
enhancement in the growth of plants (Nayak et al., 2015). The contents
of heavy metals followed the order Fe > Mn > Cr > Ni > Cu >
Zn > Co > Pb > Cd in the FA as presented in Table 3. The con-
centration of Fe was found higher as compared to other metals, i.e.,
489.7 mg kg−1 followed by Mn with concentration of 86.39 mg kg−1.
The incorporation of FA in higher amount may pose detrimental effects
to the soil, plants, and the overall environment due to its pozzolanic
effect (Jala and Goyal, 2006).

3.2. Physico-chemical characteristics of matured compost

3.2.1. pH
The variation of pH in different vessels during the process of com-
posting is shown in Fig. 1(A). The value of pH in all the vessels during
the process of composting was recorded in the range 5.25–8.15 which
indicates that at the initial stage, the pH values decreased due to the
production of organic acids by fermentation of carbohydrates, and
subsequently, the pH in all vessels was found to be increased as a result
of release of ammonia to the proteolytic activities of microbes (Pathak
et al., 2012). The analysis of the compost sample in all the vessels at
different stages showed that the pH value increased until the 15th day
of composting. The increment in pH value during composting indicates
the decomposition of organic matter into ammonium ions (Zakarya
et al., 2018). After the 15th day, the highest value for pH of 8.15 was
observed in Vessel 10 followed by Vessel 7 having pH 7.85, Vessel 6
having pH 7.80 and Vessel 4 having pH 7.75, respectively and further,
the pH values were observed to be constant till maturity. From the
initiation of the experiment until the 8th day (thermophilic stage), drop
in pH indicates the accumulation of organic acids resulting due to the
degradation of organic matter (Zakarya et al., 2018).

3.2.2. Temperature
The highest temperature was recorded on the 8th day in all the
vessels, as shown in Fig. 1(B). This initial increment in temperature was
the result of microbial activity in a thermophilic stage, which leads to
the decomposition of cellulose, hemicellulose, and lignin contents
present in organic waste materials. By comparing the temperature re-
corded in all the vessels, as shown in Fig. 1(B), it was observed that the
highest temperature of 48.50 °C was recorded in vessel 10 followed by
Vessel 7 having temperature 47 °C, Vessel 4 having temperature 45.50
°C and Vessel 9 with a temperature of 43 °C on the 8th day. The ambient
temperature measured during the composting process was 29 °C, while
Fig. 1. Variation of temperature (A) and pH (B) in vessels with different sub-
strate combination (results are the mean of three replicates and error bars in-
dicates standard deviation).
the temperature recorded in all the vessels was in the range
29.50–48.50 °C. During the thermophilic stage, the lowest temperature
of 38.5 °C was recorded on the 8th day in Vessel 1, which may be due to
the absence of jaggery and KW. In the present study, the jaggery was
used as an activator, which can be considered as an additional carbon
source resulting in the enhancement of microbial activity in the

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A. Mandpe, et al. Bioresource Technology 293 (2019) 122047

composting process. From the observations in all the vessels, the

highest microbial activity was found in Vessel 10 with the highest
temperature in the thermophilic stage. The decrease in temperature was
observed in all the vessels from the 8th day till maturity indicating the
gradual decrease in microbial activity with time due to the scarcity of
the organic matter in the composting system. Therefore, in the cooling
phase, the microbial activity in all the vessels tend to cease (Zakarya
et al., 2018).

3.2.3. Colour, odour, and texture

The compost in vessels 1 and 2 had a dark brown colour, while the
vessels 3, 4, and 6 had a dark brownish colour. Compost in vessels 5, 7,
8, 9 and 10 exhibited greyish colour which may be due to the presence
of FA in the system. Vessels 1 and 2 had a faint odour, and the vessels
3–8 had an earthy odour, while vessels 9 and 10 had a putrefying odour
which may be due to the moisture present in the system even after the
maturity of the compost. Compost in vessels 9 and 10 displayed a
wringing texture, whereas compost in other vessels had texture similar
to soil (after sieving).

3.2.4. Carbon, nitrogen, potassium, and phosphorus content

The concentrations of carbon (C), nitrogen (N), potassium (K) and
phosphorus (P) in the compost samples are presented in Table 4. It has
been perceived that the amount of total carbon (TC) ranged between
15.07% and 30.1% in different vessels, whereas, the least proportion of
TC, i.e., 15.07% was determined in the compost sample of Vessel 5, and
the highest value of 30.1% for TC was determined in the compost
sample of Vessel 1. Similar results were reported by Batjes (1996),
where the optimum value for TC was recorded more than 10%.
The total nitrogen content for the compost samples in all the vessels
was detected in the range 0.63% to 2.05%. The compost in Vessel 5
exhibited the least amount of total nitrogen content, whereas, the
highest value of total nitrogen content was recorded for the compost
sample in Vessel 1. The results are in agreement with the results ob-
tained by Khater (2015), where the amount of total nitrogen was ob-
served between 0.95 and 1.68%.
The C:N ratio of the compost is a fundamental component which Fig. 2. Analysis of heavy (A) and essential metals (B) in different vessels on
should be less than or equal to 25 after maturity in an ideal ready to use maturity of compost (results are the mean of three replicates and error bars
compost (Khater, 2015). The C:N ratio determines the amount of mi- indicates standard deviation).
crobial activity in compost taking place during the process of com-
posting. The C:N ratio of the samples in all the vessels was found to be The maximum phosphorus content was observed in Vessels 2, 6, 7, 9
in the range 20.77–28.18. The carbon and nitrogen are the building and 10 due to the presence of FA and jaggery while the least amount,
blocks of the cell structure, and when the nitrogen or carbon contents i.e., 0.05% was found in vessel 8. The phosphorus concentration in all
are found in a meagre amount in the compost, then it can be concluded the vessels ranged between 0.05% and 0.92%, as presented in Table 4.
that the microbes have utilized these contents completely (Manyapu Also, the highest potassium content was recorded for Vessel 10 with a
et al., 2018). In the present study, the highest value of C:N ratio of value of 0.04%, followed by Vessels 2, 6, and 7. The higher amount of
28.18 was observed in Vessel 5 followed by Vessel 2 with C:N ratio of potassium in the respective vessels is due to the presence of jaggery,
25.86 and Vessel 1 having C:N ration of 25.70, whereas value of C:N which contained 1056 mg of potassium per 100 g (Manyapu et al.,
ratio less than 25 was observed in Vessel 10 having value of 20.77 2018). The amount of potassium in all the vessels was detected less than
followed by Vessel 7 with C:N value of 21.59 and vessel 4 having C:N 1% which is within permissible limits as per the guidelines of Central
value of 22.64.

Table 4
Physico-chemical characterization of matured compost in different vessels.
Sr. No. Vessel TC (%) TN (%) C:N S (%) P (%) K (%)

1. V1 30.10 ± 0.3 2.05 ± 0.03 25.70 ± 0.6 0.09 ± 0.02 0.61 ± 0.006 0.03 ± 0.001
2. V2 28.04 ± 0.555 1.09 ± 0.01 25.86 ± 0.3 0.24 ± 0.001 0.91 ± 0.02 0.04 ± 0.001
3. V3 27.26 ± 0.26 0.98 ± 0.01 23.23 ± 0.41 0.21 ± 0.002 0.34 ± 0.002 0.03 ± 0.001
4. V4 23.59 ± 0.37 0.98 ± 0.003 22.64 ± 0.3 0.19 ± 0.002 0.25 ± 0.002 0.01 ± 0.007
5. V5 15.07 ± 0.2 0.63 ± 0.005 28.18 ± 0.4 0.09 ± 0.005 0.21 ± 0.004 0.02 ± 0.001
6. V6 21.56 ± 0.3 1.08 ± 0.004 23.24 ± 0.5 0.22 ± 0.002 0.71 ± 0.006 0.03 ± 0.001
7. V7 17.21 ± 0.11 1.04 ± 0.005 21.59 ± 0.6 0.16 ± 0.003 0.09 ± 0.002 0.04 ± 0.001
8. V8 16.17 ± 0.2 0.82 ± 0.004 24.75 ± 0.6 0.15 ± 0.002 0.05 ± 0.002 0.03 ± 0.001
9. V9 22.41 ± 0.4 1.09 ± 0.05 22.20 ± 0.4 0.23 ± 0.004 0.07 ± 0.001 0.02 ± 0.001
10. V10 22.84 ± 0.4 1.08 ± 0.005 20.77 ± 0.43 0.25 ± 0.003 0.92 ± 0.003 0.04 ± 0.001

Columns represent mean of three replications and standard error bars.

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A. Mandpe, et al. Bioresource Technology 293 (2019) 122047

Fig. 3(A). The differences observed for metal concentration in the ma-
tured compost reveal the different cellular mechanism of metal accu-
mulation in microbial cells that may control the translocation of metals
in the plants (Ã et al., 2007; Hanc et al., 2014; Manu et al., 2017). The
bioavailability and bioaccumulation of metals depend upon several
factors, such as mineralogy, pH, humic substances, texture, and che-
mical speciation (Dias et al., 2010; Nayak et al., 2015).

3.3. Effect of FA on microbial population of compost

The development of FA composting strategies with organic waste

Fig. 3. Enhanced percent of microbial population and enzymatic activity in
different vessels with different substrate combinations over control soil.
Public Health and Environment Engineering Organization (CPHEEO),
Government of India (2000).
3.2.5. Heavy metals content
The results of the heavy metal analysis are shown in Fig. 2(A) and
(B), respectively. The amount of diethylenetriamine penta-acetate
(DTPA) extractable metals in the compost samples of all the vessels
after maturity were found to have decreased in concentrations in
comparison to the heavy metals in the waste’s substrate. Also, the metal
concentrations recorded in all the vessels were within the permissible
limit (Awasthi and Bhatnagar, 2000). The results of the heavy metal
analysis in the present study displayed that an adequate amount of FA
amendment during the composting process did not lead to increase the
concentration of heavy metals in matured compost as shown in
Fig. 2(A). It was found that the matured compost in Vessel 10 contained
a minimum concentration of heavy metals in comparison to other
vessels, as shown in Fig. 2(A) and (B). The FA used in this study had a
neutral pH. Thus, the decreased level of DTPA extractable metals in
compost could be due to their lower concentration in FA. Heavy metals
concentration in all vessels were found to be in the order Cu >
Ba > Mn > Zn > Cr > Ag > Ni > Li > Pb whereas, the con-
centration of essential micro and macro elements were observed in the
order Ca > Fe > K > Mg > Al > Na as shown in Fig. 2(B).
In comparison to other trace metals, the concentrations of Ca, Fe
and Cu were found to be higher, whereas the concentrations of Cd, Li,
Ag, Pb, and Ni were detected in a negligible amount as shown in
can be a cost-effective and sustainable method for reduction of toxic
elements in FA through bio-adsorption and can improve the bioavail-
ability of plant nutrients which may lead to significant enhancement of
the nutrient source for plants. FA contains only 0–0.2% of Nitrogen (N)
and 0–0.34% Carbon (C), which makes it an inorganic product (Jala
and Goyal, 2006) and unable to promote the microbial activity. Bio-
logical decomposition of FA can be achieved by adding the rich and
balanced carbon source into the compost (Anbalagan and Manivannan,
2012). Thus, in the present study, FA was composted with organic
waste substrate and jaggery as an additional carbon and nitrogen source
to ascertain its effect on the rapid composting process. The microbial
activity in the soil and compost can be considered as its quality in-
dicator (Nielsen and Winding, 2002) and the application of FA was
found to enhance the microbial activity in all the combinations. The
microbial populations estimated by CFU count with different FA
amendments in compost at different time intervals are presented in
Table 5. The maximum microbial population of 342.67 CFU 108/g was
observed in Vessel 10 followed by Vessel 7 with 325.33 CFU 108/g,
Vessel 9 with 287.00 CFU 108/g, Vessel 4 with 256.67 CFU 108/g,
Vessel 6 with 225.00 CFU 108/g and Vessel 3 having 216.67 CFU 108/g
in comparison to the control as shown in Fig. 3. The enhancement in the
bacterial population of the compost samples in Vessels 3, 4, 6, 7, 9 and
10 might be due to the neutralizing effect of FA on soil (Surridge et al.,
2009) which could have led to increasing the mobility of nutrients in
the compost. A reduction in the bacterial population was observed in
Vessel 8 with 196.33 CFU 108/g and Vessel 5 with 173.00 CFU 108/g as
presented in Table 5. This diminution in the bacterial count of the
compost samples is an attribute of the higher dose of FA, which served
the high availability of heavy metals to the compost.
3.4. Influence of FA amendment on dehydrogenase activity of compost
Enzymatic activity is a direct attribute of the microbial community
for its metabolic requirements (Waghmare et al., 2014; Nayak et al.,
2015) and thus, it is an indicator of the good quality of soil and compost
(Caldwell, 2005). The oxidation of organic compounds is a dehy-
drogenation action of specific dehydrogenase enzymes (Burns, 1978).

Table 5
Effect of fly ash amendment on microbial and enzymatic activity of compost.
Sr. No. Treatment Microbial activity (CFU 108 g−1) Dehydrogenase activity (μg TPF/g−1/h)

8 Days 15 Days Maturity 8 Days 15 Days Maturity

1. V1 114.67 ± 14.29 95.67 ± 13.32 78.00 ± 12.12 4.19 ± 0.32 3.45 ± 0.17 1.86 ± 0.10
2. V2 148.00 ± 9.54 139.67 ± 9.07 132.67 ± 10.07 5.33 ± 0.40 3.70 ± 0.21 2.04 ± 0.06
3. V3 216.67 ± 6.11 206.33 ± 8.08 196.00 ± 9.54 8.29 ± 0.70 5.51 ± 0.41 3.78 ± 0.16
4. V4 256.67 ± 9.29 247.67 ± 9.29 240.67 ± 10.26 11.26 ± 0.70 6.48 ± 0.40 4.40 ± 0.47
5. V5 173.00 ± 8.54 160.00 ± 8.54 145.67 ± 7.77 6.52 ± 0.51 4.44 ± 0.36 2.62 ± 0.51
6. V6 225.00 ± 5.57 218.67 ± 8.50 211.67 ± 7.77 11.02 ± 0.55 6.47 ± 0.46 4.08 ± 0.10
7. V7 325.33 ± 14.19 311.67 ± 13.65 300.00 ± 13.11 14.70 ± 1.15 8.58 ± 0.52 6.56 ± 0.37
8. V8 196.33 ± 12.01 191.67 ± 11.06 186.00 ± 11.53 7.58 ± 0.51 4.60 ± 0.35 2.81 ± 0.18
9. V9 287.00 ± 11.00 275.67 ± 11.50 265.00 ± 11.00 12.93 ± 0.43 7.34 ± 0.28 5.52 ± 0.37
10. V10 342.67 ± 15.63 333.00 ± 16.70 325.00 ± 12.17 16.14 ± 1.33 10.89 ± 0.12 8.56 ± 0.53
11. Soil 95.00 ± 9.54 87.33 ± 9.50 85.00 ± 7.55 3.67 ± 0.57 2.94 ± 0.09 1.87 ± 0.17

Columns represent mean of three replications and standard error bars.

6
 A. Mandpe, et al.

Table 6
Correlation analysis of microbial activity, heavy metals, temperature and pH measured during composting of different FA, OW, KW and additive combinations.
Parameters Microbial Ag Al Ba Ca Cd Cr Cu Fe K Li Mg Mn Na Ni Pb Sr Zn Temperature pH
activity

Microbial 1.000 – – – – – – – – – – – – – – – – – – –
activity
Ag −0.577* 1.000 – – – – – – – – – – – – – – – – – –
Al 0.751** −0.436 1.000 – – – – – – – – – – – – – – – – –
Ba −0.627* 0.099 −0.844** 1.000 – – – – – – – – – – – – – – – –
Ca −0.524* 0.050 −0.830** 0.882** 1.000 – – – – – – – – – – – – – – –
Cd −0.293 −0.003 −0.245 −0.087 0.177 1.000 – – – – – – – – – – – – – –
Cr −0.253 0.462 −0.235 0.200 0.386 −0.029 1.000 – – – – – – – – – – – – –
Cu 0.114 −0.273 0.591* −0.313 −0.356 −0.203 −0.009 1.000 – – – – – – – – – – – –

7
Fe −0.593* 0.127 −0.829** 0.810** 0.869** 0.338 0.312 −0.568* 1.000 – – – – – – – – – – –
K −0.548* 0.152 −0.795** 0.819** 0.930** 0.212 0.539* −0.392 0.868** 1.000 – – – – – – – – – –
Li 0.058 0.564* 0.153 −0.409 −0.401 0.025 0.453 −0.096 −0.236 −0.264 1.000 – – – – – – – – –
Mg −0.526* 0.040 −0.801** 0.771** 0.940** 0.435 0.393 −0.455 0.945** 0.930** −0.288 1.000 – – – – – – – –
Mn −0.345 0.087 −0.554* 0.421 0.731** 0.566* 0.522* −0.318 0.743** 0.668** 0.039 0.840** 1.000 – – – – – – –
Na −0.059 −0.127 −0.338 0.577* 0.509* −0.234 0.346 −0.247 0.478 0.692** −0.213 0.488* 0.103 1.000 – – – – – –
Ni −0.694** 0.263 −0.773** 0.849** 0.891** 0.023 0.509* −0.222 0.784** 0.908** −0.372 0.806** 0.532* 0.523* 1.000 – – – – –
Pb −0.121 0.174 0.016 0.104 0.261 −0.270 0.578* 0.497 −0.066 0.131 0.049 0.073 0.312 −0.181 0.340 1.000 – – – –
Sr −0.574* 0.027 −0.813** 0.717** 0.905** 0.568* 0.252 −0.412 0.871** 0.869** −0.372 0.962** 0.799** 0.349 0.759** 0.036 1.000 – – –
Zn −0.695** 0.220 −0.761** 0.838** 0.904** 0.123 0.599* −0.118 0.777** 0.895** −0.200 0.840** 0.678** 0.456 0.928** 0.442 0.791** 1.000 – –
Temperature 0.919** −0.375 0.598* −0.487 −0.367 −0.405 −0.058 0.088 −0.527* −0.370 0.079 −0.428 −0.294 0.115 −0.513* 0.022 −0.500* −0.549* 1.000 –
pH 0.845** −0.458 0.598* −0.610* −0.390 0.019 −0.206 0.008 −0.446 −0.334 −0.078 −0.323 −0.220 0.048 −0.529* −0.244 −0.319 −0.625* −0.854** 1.000

Significance levels for positive correlations are *p ≤ 0.05 (significant) and **p ≤ 0.01 (highly significant).
Bioresource Technology 293 (2019) 122047
A. Mandpe, et al.
Fig. 4. PCA of various parameters (TC, TN, C/N, temperature, pH, moisture
content, heavy metals, microbial & dehydrogenase activity) for matured com-
post produced from composting of FA with different OW, KW and additive
combinations.
The dehydrogenase activity is an estimate of biological metabolism, and
hence, it is a potential indicator of compost quality, which may further
maintain the fertility of the soil (Makoi and Ndakidemi, 2008). The
results of the present study indicated that the application of FA in
compost significantly enhanced the dehydrogenase activity in all the
vessels over the control soil (3.67 μg TPF/g−1/h) as shown in Fig. 3.
Composting of only garden waste seemed to have not much effect on
the enzymatic activity and was observed to increase slightly in Vessels 1
and 2 as 5.33 and 4.19 μg TPF/g−1/h, respectively over the control as
presented in Table 5. The maximum dehydrogenase activity was re-
corded on the 8th day in Vessel 10 with a value of 16.14 μg TPF/g−1/h
amended with 20% FA and 50 g jaggery followed by Vessel 7 with a
value of 14.70 μg TPF/g−1/h having a combination of 20% FA with
150 g jaggery. These beneficial effects might have been observed due to
the supply of nutrients by the FA for the necessary activity of dehy-
drogenase enzymes which also aided the essential nutrients in the
compost and created a conducive environment for the enzymatic ac-
tivity (Pati and Sahu, 2004). On the other hand, the higher dose (30%)
of FA in compost was found to inhibit the activity of compost enzymes
as recorded in Vessels 5 and 8 with a value of 6.52 μg TPF/g−1/h 5 and
7.58 μg TPF/g−1/h, respectively. However, the recorded enzymatic
activity at the higher dose of FA was better than the compost of organic
waste alone and the control soil. Similar results for nutrient availability
in plants were reported by Tejada and González, (2009).
3.5. Inter-relationship between different parameters
The data recorded in the microbial study and the physico-chemical
characterization of compost was analyzed using the correlation analysis
as presented in Table 6. The correlation analysis showed a significant
positive correlation between the microbial activity, pH and the tem-
perature of the compost while a significant negative correlation was
observed for microbial activity and heavy metals concentration in all
the vessels as presented in Table 6. Also, the pH and temperature were
found to have a strong negative correlation with each other. A similar
relation between pH and microbial activity was also found by Yun et al.
(2016). Principal Component Analysis (PCA) of all the ten vessels was
carried out to determine the preeminent combination of applied sub-
strates on the basis of twelve different parameters like TC, TN, C/N, S%,
P%, K%, microbial activity and dehydrogenase activity on 8th day, 15th
day and at the time of maturity, respectively as shown in Fig. 4. In the
present study, the effect of heavy metals concentration in FA on the
microbial and enzymatic activity in the composting system was
8
Bioresource Technology 293 (2019) 122047
investigated, and effect of different combinations of organic waste
substrate & FA on all the parameters was also ascertained. The first and
second component accounted for 7.198 and 2.477 eigenvalues, re-
spectively.
The substrate combination of vessel 10 was seen separated from the
rest of the vessels combinations in the score plot as shown in Fig. 4 and
unambiguously justifies the best compostable composition of the stu-
died substrates. Apart from vessel 10, vessels 1, 2 and 5 also had sig-
nificant separation from the rest of the vessels and are in line with the
findings of Escobar and Solarte (2015) as they have also reported si-
milar results in chemical and microbiological variables.
4. Conclusion
Substrate combination of compost comprising 50% BW, 30% KW,
20% FA, and 50 g jaggery exhibited maximum microbial and enzymatic
activity with a good compost quality. However, there is a need to
evaluate and scale up these experimental findings on pilot and field
level. Also, emphasis is needed to study and explore the consequences
of enduring use of this compost on soil quality and plant growth.
Further, to ascertain the effectiveness of FA on microbial species and
disinfection of pathogenic microorganism during the composting pro-
cess, metagenomics analysis of compost samples is required to be done
which is in the further scope of the study.
Acknowledgment
The authors acknowledge the Director, CSIR-NEERI for facilitating
the activities & also giving permission to publish this research work.
Declaration of Competing Interest
No Interest.
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