TOPIC

Sabjiyon Ke Sukshma Poshak Tatvon Par

Rasaynik Avam Jaivik Khad Ke Prabhav
Ka Addhyyan
INTRODUCTION

Studies have indicated the effects of chemical and organic manure on the
micronutrient of vegetables. biofertilizers on micronutrients, growth, fruit yield and
quality in solanaceous and okra crops. The chemical fertilizers are not only in
minimum supply, but also they are costly in developing countries like India.
Moreover, continuous use of chemical fertilizer affects the soil health and lead to
the environmental hazards. Therefore, by using the organic manure to supplement
part of the nutrient needs of the plant, not only the cost of inputs be brought down,
but also the environmental hazards associated with chemical fertilizers can be
avoide.

The usage of organic manure adds soil with crucial nutrients and adsorbs nutrients
against leaching (Follett et al., 1981). As a result it also improves the soil fertility
and its texture along with enhanced microbial population, ion exchange capacity of
soil and moisture holding capacity of soil (Arancon et al., 2005). At the same time,
usage of organic manure reduces pollution rate in the soil, water and in the
produced fruits (El-Agory et al., 1996). Often organic manure contain one or more
of symbiotic and/or non-symbiotic N-fixing bacteria or phosphorus dissolving
bacteria or potassium mobilizing bacteria (Shaheen et al., 2016).
Plant Micronutrients
Plants, like all other living things, need food for their growth and development. Plants
require 16 essential elements. Carbon, hydrogen, and oxygen are derived from the
atmosphere and soil water. The remaining 13 essential elements (nitrogen,
phosphorus, potassium, calcium, magnesium, sulfur, iron, zinc, manganese, copper,
boron, molybdenum, and chlorine) are supplied either from soil minerals and soil
organic matter or by organic or inorganic fertilizers (R. Uchida, 2000). Other
elements that are needed by plants in much lower amounts than the macronutrients
are called micronutrients or trace elements. They are as essential to plant growth as
the macronutrients because they perform very essential and specific roles, particularly
in molecules involved with energy transfer process, hormones, and enzymes.
Role of plant Micronutrient:
Micronutrients are needed in very small amounts. Their adequate concentrations in
plants are generally below the 100 parts per million (ppm) level (Table 1). The
essential micronutrients are zinc (Zn), iron (Fe), manganese (Mn), boron (B), chlorine
(Cl), copper (Cu), molybdenum (Mo), cobalt (Co), vanadium (V), sodium (Na), and
silicon (Si).
A primary function of boron is related to cell wall formation, so boron-deficient
plants may be stunted. Sugar transport in plants, flower retention and pollen
formation and germination also are affected by boron. Copper is necessary for
carbohydrate and nitrogen metabolism and, inadequate copper results in stunting of
plants. Iron is involved in the production of chlorophyll, and iron chlorosis is easily
recognized on iron-sensitive crops growing on calcareous soils. Iron also is a
component of many enzymes associated with energy transfer, nitrogen reduction
and fixation, and lignin formation. Manganese is necessary in photosynthesis,
nitrogen metabolism and to form other compounds required for plant metabolism.
Molybdenum is involved in enzyme systems relating to nitrogen fixation by
bacteria growing symbiotically with legumes. Zinc is an essential component of
various enzyme systems for energy production, protein synthesis, and growth
regulation. Because chloride is a mobile anion in plants, most of its functions relate
to salt effects (stomatal opening) and electrical charge balance in physiological
functions in plants (Raun Lohry, 2007).
 Figure -1.1 Role of Micronutrient in plant

Organic farming is a new technique of agricultural production system which includes

naturally and locally available organic materials or agro inputs to meet out the
production system without harming our treasured natural resources.

Organic manure has significant aspect in plant growth as a source of all necessary
micro and macronutrients in accessible forms throughout mineralization and
bettering chemical and physical properties of soils (Hala and Nadia, 2009).
In the prior 50 years history in India, the chemical fertilizers and pesticides have
dedicated an immense appearance in the increment of agricultural productivity.
Aimless operations of chemical pesticides and fertilizers furnished in decline of soil
productivity along with adverse effects on human health (Roychowdhury et al., 2014)

Phytochemicals

Phytochemicals are the concoctions secreted by different parts (bark, leaves, flower,
roots and seeds) of a plant.

Fruits and vegetables are advised to be safe for health of humans. This property is
associated with chemical composition of these foods, specially to phytochemicals
(known as bioactive compounds), which comprises flavonols, flavones, phenolic
acids, isoflavones and glucosinolates. These phytochemicals convey the impression
contrary to progression of distinct types of cancer and cardiovascular diseases, since
these compounds could administer anti-inflammation properties, antioxidant capacity
(AOC), antitumor effects and lipid profile modification. Phytochemicals are
metabolites of which there are several classes including; alkaloids, glycosides,
flavonoids, gums, phenols, polysaccharides, tannins, phlobatannins, terpens and
terpenoids (Mensah et al., 2013).
AIM
Study the effect of chemical and organic manure on micronutrients of vegetables.

OBJECTIVES:
• To assess the potential of the earthworm species (Eisenia foetida) to transform
agricultural waste into useful compost.
• To evaluate the physicochemical parameters of the soil.
• To evaluate the physicochemical parameters of the vermicompost produced.
• To evaluate the effect of (organic manure) vermicompost on vegetable crops as
compare to chemical fertilizers.
• To evaluate the phytochemical and antioxidant activity of vegetable crops.
 MATERIALS AND METHOD
Soil Sample Collection:
The aim of this study was to determine the physicochemical status of soil and
vermicompost samples. Before sampling 15 mm top soil was removed. Soil samples
were collected from different selected agriculture field at the depth of 15cm. A
composite sample of about 500 g was taken through mixing of represented soil sample.
These soils were first sieved by scientific siever . The sieved out fine particles are then
dried at room temperature for several hourshours. The samples were carefully labeled
and packaged.

Vermicompost preparation:
In this study, earthworm feed, mixture of plant
residue, grass and kitchen waste residue mix with
grass and cow dung and temple waste for
vermicompost preparation. All residues were
chopped into small pieces (1 cm per piece) before
feeding on the bedding. Fresh wastes were fed
every 5 days for each condition. Than the 6 month
old earthworms were selected. The length,
thickness and body weight were measured.
Physico-chemical analysis of soil and vermicompost:
Samples were analyzed for physicochemical parameters such as pH, EC, Nitrogen, organic carbon,
bulk density, colour, moisture, phosphorous content, and organic matter and micronutrients by
employing standard methods (Trivedy and Goel, 1984).
pH analysis:
The pH meter was calibrated by using 4.01, 6.86, 9.18 buffer solution. The pH of 1:5 Soil: deionised
water suspension was determined by calibrated pH meter at room temperature.
Bulk density:
Bulk density was determined by ratio of oven dry mass of the soil to bulk volume of the soil using
bulk density apparatus.
Moisture content
Moisture content was determined by ratio of the weight of water in the soil to weight of the dry soil.
The ratio was expressed as percentage. The percentage moisture content was corrected by using
moisture correction factor (MCF).6

Organic carbon
Soil organic carbon was estimated, following titration method of Walkley and Black (1934) by
Tiwari et al., 1987. To 5 g of oven dried mine spoil, 10 ml of 1 N K 2Cr2O7 and 20 ml of conc. H2SO4
were added in a 500 ml flask, thoroughly shaken for 5 min, and was allowed to stand for 30 min.
The suspension was diluted with 200 ml of distilled water, followed by 1 ml of 85% H 3PO4, and 1
ml of diphenylamine indicator. The mixture was titrated against 1N (NH 4)2Fe(SO4)2.6H2O, until the
colour of the mixture flashed to green. Then, 0.5 ml of 1N K 2Cr2O7 was added, and the titration was
completed by adding 1N (NH4)2 Fe(SO4)2.6H2O, till the last traces of blue color disappeared.
Organic carbon (%) was calculated as: [(V1-V2)/W]×0.003×100
Where V1=volume of 1 N K2Cr2O7
V2=volume of 1 N (NH4)2Fe(SO4)2.6H2O
W=wt of soil sample.
Total Nitrogen
Total soil nitrogen was determined by Kjeldahl method. Spoil sample of 10 g was
transferred to a 300 ml Kjeldahl flask and was moistened with 25 ml of distilled water;
allowed to stand for 30 min. To it, 20 g of sodium sulfate and the catalyst mixture (20 g
copper sulfate, 3 g of mercuric oxide, 1 g selenium power were ground) was added. To one
part of this mixture, 20 parts of anhydrous sodium sulfate was added, and a pinch of
granulated zinc was added to the suspension, followed by 35 ml of conc. H2SO4. The
resulting mixture was subjected to low heat treatment for 30 min for digestion, till the digest
become yellow and colorless. The digest was then cooled and 100 ml of water was added,
and allowed to stand for 5 min. The supernatant was then transferred into a flask. 25 ml of
4% boric acid was pipetted into a 500 ml conical flask, and 5 drops of mixed indicator (0.5 g
bromo-cresol green and 0.1 g methyl red dissolved in 100 ml of 95% ethyl alcohol) was
added. The glass tube attached to the lower end of the condenser as dipped into the boric
acid solution. The condenser was connected to the flask and 100 ml of 40% NaOH was
added slowly through the separating funnel. By heating the mixture, 150 ml of distillate was
collected in the conical flask and was titrated against N/14 H2SO4, till the faint pink
coloration was reached. A blank was run instead of soil.
Total soil nitrogen (%) was calculated as: [(T-B)×N×14.007×100]/W;
where T and B are the volume of titrant used against sample and blank;
N=normality of titrant and W=weight of sample.
Soil texture
Soil texture analysis included the estimation of clay (<0.002 mm), silt (0.06 mm-0.002 mm) and
sand (2 mm-0.06 mm) percentage. Soil sample (50 g) was taken in a 500 ml heat resistant bottle,
calibrated up to 250 ml. To this, 125 ml of water was added and the mixture was swirled to wet
the spoil thoroughly. 20 ml of 30% hydrogen peroxide was added to it and the bottle was gently
rotated. Few drops of amyl alcohol were added to the mixture and kept in a boiling water bath,
till the reaction was complete. Then, 2 g of sodium hexametaphosphate was added, followed by
water, to make it up to 250 ml, and was shaken for 28 hr in a mechanical shaker. Then, the
contents were transferred to a 1 L sedimentation cylinder and the volume was made up to 1 L. A
blank cylinder was maintained by dissolving 2 g of sodium hexametaphosphate in water and
made up to the mark with water. Both experimental and blank samples were placed in a water
bath to maintain a constant temperature (25 ± 2°C). After 30 min, the sample cylinder was mixed
vigorously with a plunger. The Bouyoucos hydrometer readings were taken exactly at 40 sec and
5 hr for the samples, and at 5 hr for the blank. Temperature of the water bath was recorded. The
percentage of sand, silt and clay were determined as per the following calculation.
Phosphorus analysis.
Water extracted P from soils were analyzed by ascorbic acid colorimetry (Murphy
Riley) method. To prepare samples, 4.0 mL Reagent B and 19.0 mL Distilled water was
added to 2.0 mL of each extract. Standards consisting of 5.0 mL of each standard P
solution (0.1 ppm to 1.0 ppm P), 4.0 mL Reagent B, and 16.0 mL DI water and a 0.0
ppm P standard consisting of 4.0 mL Reagent B and 21.0 mL DI water were also
prepared. Samples were allowed 30 minutes for color development.
Vermicompost Performance Test on Vegetable Crop:
The vermicompost performance was investigated in Brinjal and Ladyfinger plants. Both
plant seeds were planted in 10 cm of diameter pot. Experiment was conducted in
Randomized Complete Blocks Design (RCBD) with four replication and four treatment
(T1 = Control, T2 = Vermicompost, and T3 = Chemical) at Rapture biotech Institute,
Bhopal during 2017. The material consisted of one brinjal and ladyfinger vegetables.
The organic manure used was vermicompost. The seeds sown in the rainy season and
both plants were grown in four row plots, each plot included two ridges and each ridge
was 2.5 m in length and 50 cm apart. Vermicompost was applied by 500 g every 10 days
after seed germination in condition 1, 2 and 3. In the 4th condition, chemical fertilizer
was applied by 20 g every 10 days after seed germination. Each condition was repeated
for three times. All plants were cultivated in nursery with 50% of actual light intensity.
After 56 day of plant cultivation, these plants were harvested.
Morphological characteristic analysis:
An Agronomic characteristics included plant height, number of fruit per plant,
individual fruit weight per plant, total fruit weight per plant, fruit size and fruit yield
per green house. Data were recorded on 4 competitive plants of each plot was
calculated for the entire plot.
Physico-Chemical Analysis of plants:
The physico-chemical analysis includes number of parameters such as physical state,
colour, taste, percentage of loss on drying as per standard method of (The Indian
Pharmacopoeia, 1996), ash content as per method of (Gupta, 2003 and Indrayan et al.,
2005 ), ash value (water, alcohol and acid soluble or insoluble ash) as per method of
(Ahmad and Sharma, 2001), pH value and conductivity (Sharma, and Kaur, 1998),
Chloride and Sulphate. The eight inorganic elements (Ca, P, Mn, Zn, Ni, Fe, K and Mg)
were detected in plant materials (Archa Vermani et al., 2010).
Phytochemical analysis of vegetable plants:
The extracts were subjected to qualitative tests for detection of phytoconstituents
present in it viz. alkaloids, carbohydrates, glycosides, phytosterols, fixed oils & fats,
phenolic compounds & tannins, proteins, flavanoids, lignins and saponins (Harborne,
1984).
PHYTOCHEMICAL INVESTIGATION
1) Test for Alkaloids
Methanolic extract was warmed with 2% H2SO4 for two minutes. It is filtered and a few
drops of reagents were added and indicated the presence of alkaloids.
a. Mayer’s reagent-A creamy- white colored precipitation positive.
b. Wagner’s reagent-A reddish-brown precipitation positive.
2) Test for Flavonoids
A small quantity of the extracts is heated with 10 ml of ethyl acetate in boiling water for 3
minutes. The mixture is filtered differently and the filtrates are used for the following test.
A. Ammonium Test
The filtrate was shaken with 1 ml of dilute ammonia solution (1%). The layers were
allowed to separate. A yellow coloration was observed at ammonia layer. This indicates the
presence of the flavonoid.
B. Aluminum Chloride Test
The filtrates were shaken with 1 ml of 1% aluminum chloride solution and observed for
light yellow color. It indicated the presence of flavonoid and diluted NaOH and HCl was
added. A yellow solution that turns colorless indicated positive.
3) Test for Terpenoids
Salkowski Test
The extract was mixed with 2ml of chloroform and concentrate H 2SO4 (3ml) is
carefully added to form a layer. A reddish brown coloration of the interface is formed
to show positive result of the presence of terprnoids.
4) Test for Phenol /Tannin
A small quantity of the extract is boiled with 5 ml of 45% solution ethanol for 5
minutes. Each of the mixture is cooled and filtered. The different filtrates were used
to the following test:
b) Lead Sub Acetate Test
1ml of the different filtrate was added with three drops of lead sub acetate solution.
A cream gelatinous precipitation indicates positive test for Tannins.
c) Ferric Chloride Test
1ml each of filtrate is diluted with distilled water and added with two drops of ferric
chloride. A transient greenish to black color indicates the presence of Tannins.
5) Test for Steroids
2ml of acetic anhydride was added to extract 2ml of H 2SO4. The color changed from
violet to blue or green in some samples indicated the presence of steroids.
6) Test for Saponins
Frothing Test
A small quantity of different extract was diluted with 4 ml of distilled water. The
mixture was shaken vigorously and then observed on standing for stable brake.
7) Test for Carbohydrates
The extract was shaken vigorously with water and then filtered. To the aqueous
filtrate was added few drops of Molisch’s reagents. Followed by vigorous shaking
again, concentrated H2SO4 1ml was carefully added to form a layer below the
aqueous solution. A brown ring at the interface indicated the positive.
8) Test for Proteins
5ml of distilled water was added into the extracts. This was left to stand for three
hours and then was filtered. To 2 ml portion of the filtrate was added 0.1ml Millon’s
reagent. It was shaken and kept for observation. A yellow precipitation indicates the
positive.
Determination Total Flavonoid Content
The total flavonoid content (TFC) of the extracts was determined using the aluminium
chloride assay through spectrophotometer (Samatha et al., 2012). An aliquot (0.5 ml) of
extracts was taken in different test tubes, then 2 ml of distilled water was added, followed
by the addition of 0.15 ml of sodium nitrite (5% NaNO 2, w/v) and allowed to stand for 6
min. Later 0.15 ml of aluminium trichloride (10% AlCl 3) was added and incubated for 6
min, followed by the addition of 2 ml of sodium hydroxide (NaOH, 4% w/v) and volume
was made upto the 5ml with distilled water. After 15 min of incubation the mixture turns
to pink whose absorbance was measured at 510 nm using a spectrophotometer. Distilled
water was used as blank. The TFC was expressed in mg of Quercetin equivalents (QE)
per gram of extract. All the determinations were carried out three times.
Determination of Total Phenolic Content
The total phenolic content (TPC) of the crude extracts of vegetable plants was determined
using the method of Singleton et al., (1999) with slight modifications. To 0.5 ml of test
sample, 1.5 ml (1:10 v/v diluted with distilled water) Folin Ciocalteau reagent was added
and allowed to stand for 5 min. After 5 min, 2.0ml of 7.5% of sodium carbonate was
added. These mixtures were incubated for 90 min in the dark with intermittent shaking.
After incubation, development of the blue colour was observed. Finally absorbance of
blue colour in different samples was measured at 725 nm using spectrophotometer. The
results were expressed as tannic acid equivalents TA/g of the plant material.
 PHARMACOLOGICAL EVALUATION OF PLANT MATERIALS

Total Antioxidant Activity

Total antioxidant activity was estimated by phosphomolybdenum assay (Prieto et al.,
1999). 1ml each of 0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM
ammonium molybdate were added in 20 ml of distilled water and made up volume to
50 ml by adding distilled water. All extracts of both plants in different concentration
ranging from 200 µg to 1000 µg were added to each test tube individually containing
3 ml of distilled water and 1 ml of Molybdate reagent solution. These tubes were kept
incubated at 95ºC for 90 min. After incubation, these tubes were normalized to room
temperature for 20-30 min and the absorbance of the reaction mixture was measured at
695 nm. Mean values from three independent samples were calculated for each
extract. Ascorbic acid was used as positive reference standard.
Nitric oxide scavenging activity
The procedure was based on the method, where sodium nitroprusside in aqueous
solution at physiological pH spontaneously generates nitric oxide, which interacts
with oxygen to produce nitrite ions that can be estimated using Greiss reagent.
Scavengers of nitric oxide compete with oxygen leading to reduced production of
nitrite ions. For the experiment, sodium nitroprusside (10mM) in phosphate buffered
saline (PBS) was mixed with different concentrations of all extracts dissolved in
methanol and incubated at room temperature for 150 minutes. The same reaction
mixture without the extracts but the equivalent amount of methanol served as the
control. After the incubation period, 0.5 ml of Greiss reagent [1% sulfanilamide, 2%
H3PO4 and 0.1% N-(1-naphthyl) ethylenediamine dihydrochloride] was added. The
absorbance of the chromophore formed was read at 546 nm (Garrat, 1961) using a
spectrophotometer.

Data analysis: For quantitative characters, data were analyzed for simple statistics
using the compare means and correlation analysis and regression analysis with the
help of computer software SPSS.
 RESULT

PHYSICO-CHEMICAL ANALYSIS OF SOIL AND VERMICOMPOST

The comparative results of the physicochemical analysis of soil samples and
vermicompost exhibited in Table 4.1 and Figure 4.1-4.11. Soil samples tested in the
current study were collected from eggplant (S1) and okra field (S2).
In the present study, pH ranged found between 6.8 to 7.97 (Figure 4.1). The pH of
vermicompost found slightly acidic, which is due to the enzymes and acid secreted by
earthworms.
Result on some selected physico-chemical properties of the compost and soil is
presented in Table 4.1. Conductivity values of soil samples range between 255.3 -331.8
µS/CM (Figure 4.2). Vermicompost prepared by organic matter contains the highest
(5620.6 µS/CM) electrical conductivity. It is represented by µS/CM. Electrical
conductivity (EC) expresses ion contents of solution which determine the current
carrying capacity.
In the current study, the significant amount of organic matter, total nitrogen, phosphate
and total potash was reported in the vermicompost (Table 4.1). The results of this study
revealed that the vermicompost have the highest amount of potassium compared to
nitrogen and phosphate. It was recorded in the amount of 6117.5 mg/Kg while total
nitrogen and phosphate investigated in the vermicompost is 0.072% and 360.6 mg/Kg
(Figure 4.3-4.7).
Vermicompost retains nutrients for a long time and while the conventional compost
fails to deliver the required amount of macro and micronutrients including the vital
NKP to plants in shorter time, the vermicompost does (Sinha 2009).
This study attempts to estimate the status of micronutrients in the soil sample collected
from okra and eggplant field and also in vermicompost prepared by organic matter.
Table 4.1 concluded that the soil samples collected from agro land rich in magnesium
and manganese compared to others. Magnesium content found in the range of 20.30-
26.20 mg/kg (Figure 4.9), while vermicompost prepared with various organic matter
contains Mg in the concentration of 7.94 mg/kg. Manganese was observed in the range
of 14.21-42.12 mg/Kg (Figure 4.10). Sarto et al., (2011) focused on the extraction
methods of micronutrients from soil, which is the primary step of soil analysis, while
Iatrou et al., (2015) used Mehlich 3 and DTPA tests for the determination of
micronutrients.
This study also reported the concentration of micronutrients in the vermicompost.
Vermicompost was rich with macro and micro nutrients. Calcium, manganese and zinc
found in good quantity (Table 4.1). Higher amount of micronutriest in the
vermicompost is due to the enzymatic activity of earthworms.
Vermicompost retains nutrients for a long time and while the conventional compost
fails to deliver the required amount of macro and micronutrients including the vital
NKP to plants in shorter time, the vermicompost does (Sinha 2009).
This study attempts to estimate the status of micronutrients in the soil sample
collected from okra and eggplant field and also in vermicompost prepared by
organic matter.
Table 4.1 concluded that the soil samples collected from agro land rich in
magnesium and manganese compared to others. Magnesium content found in the
range of 20.30-26.20 mg/kg (Figure 4.9), while vermicompost prepared with
various organic matter contains Mg in the concentration of 7.94 mg/kg. Manganese
was observed in the range of 14.21-42.12 mg/Kg (Figure 4.10). Sarto et al., (2011)
focused on the extraction methods of micronutrients from soil, which is the primary
step of soil analysis, while Iatrou et al., (2015) used Mehlich 3 and DTPA tests for
the determination of micronutrients.
This study also reported the concentration of micronutrients in the vermicompost.
Vermicompost was rich with macro and micro nutrients. Calcium, manganese and
zinc found in good quantity (Table 4.1). Higher amount of micronutriest in the
vermicompost is due to the enzymatic activity of earthworms.
 Table 4.1 Physico-chemical analysis of vermicompost and soil samples collected
from two different agro field

S. Parameters UNIT Sample Sample Vermi-

No. (S1) compost
(S2) 8

1 pH 7.97 7.83 6.83 7.8

2 Electrical µS/CM 255.3 331.8 5620.3 7.6

Conductivity
7.4
3 Organic matter % 1.41 2.78 23.72
7.2
4 Total Nitrogen % 0.014 0.018 0.072

pH
5 Total Phosphate Mg/Kg 2.81 10.92 360.6 7

6 Total Potass Mg/Kg 210 336 6117.5 6.8

6.6
7 Calcium Mg/Kg 20.14 12.38 35.28
6.4
8 Iron Mg/Kg 5.32 15.21 8.50
6.2
9 Magnesium Mg/Kg 20.30 26.20 7.94 sample 1 sample 2 vermi

10 Manganese Mg/Kg 14.21 42.12 18.52 SAMPLE ID

11 Zinc Mg/Kg 7.06 9.61 10 Figure 4.1 Determination of pH in both soil

samples and vermicompost
 6000 25

5000
20
4000
EC µS/CM

15

OC %
3000
10
2000

1000 5

0 0
sample 1 sample 2 vermi sample 1 sample 2 vermi
Figure 4.3 Values of organic carbon in vermicompost and
both soil samples
Figure 4.2 Electrical conductivity of vermicompost and Soil 400
sample 1 & 2.
350
0.08
300

Total Phosphate (mg/Kg)

0.07
TOTAL NITROGEN %

0.06 250
0.05
200
0.04
0.03 150
0.02
100
0.01
0 50
sample 1 sample 2 vermi
0
Figure 4.4 Values of total nitrogen in vermicompost and both sample
Figure 4.5 Values of1 total phosphate
sample 2in vermicompost
vermi and
soil samples both soil samples
 40
7000
35
POTASSIUM Mg/Kg

6000

CALCIUM Mg/Kg
30
5000
25
4000 20
3000 15
2000 10
1000 5
0 0
sample 1 sample 2 vermi sample 1 sample 2 vermi
SAMPLE ID SAMPLE ID

Figure 4.6 Values of Potassium in vermicompost and Figure 4.7 Determination of calcium in vermicompost
both soil samples and both soil samples

16
30
14

MAGNESIUM Mg/Kg
25
12
20
IRON Mg/Kg

10
15
8
10
6

4 5

2 0
sample 1 sample 2 vermi
0
sample 1 sample 2 vermi SAMPLE ID

SAMPLE ID
Figure 4.9 Determination of magnesium in vermicompost
Figure 4.8 Determination of iron in vermicompost and and both soil samples
both soil samples
 45

MANGANESE Mg/Kg
40
35
30
25
20
15
10
5
0
sample 1 sample 2 vermi
SAMPLE ID

Figure 4.10 Determination of manganese in vermicompost and both soil samples

10
9
8
7
ZINC Mg/Kg

6
5
4
3
2
1
0
sample 1 sample 2 vermi
SAMPLE ID

Figure 4.11 Determination of zinc in vermicompost and both soil samples

BIOASSAY
Comparative effect of vermicompost and chemical fertilizers on growth of okra
and eggplant:-
Okra Plant:
The results are presented in Tables 4.2 below shows the growth characteristics of the
okra grown in three distinct treatment viz., control (T1), soil treated with vermicopost
(T2) and soil treated with chemical fertilizers (T3).
The current investigation revealed that shoot length, plant height and dry weight of
okra plant was maximum in T2 treatment with 18 cm, 26.33 cm and 2.80 g,
respectively, and minimum in chemical fertilizers treated group (T3), while plant
treated with chemical fertilizers showed minimum growth in root length (4.33 cm)
which was comparatively low to vermicompost and control treatment (Table 4.2 &
Figure 4.12). The present research also studied the variable impact of vermicompost
and chemical fertilizers on fruit length and yield (Table 4.3 and Figure 4.13). In this
investigation fruit length was maximum in T2 treatment with 10.6 cm and minimum in
T1 treatment (6.51 cm), maximum fruit width was observed in T2 treatment with 2.63
cm and minimum in T3 group (0.98 cm). Average weight was maximum in T2
treatment with 15.22 gm and minimum in T1 treatment respectively. The highest yield
of okra fruits per plant recorded in T2 treatment. It was 2.83 kg/ plant (Table 4.3).
 The current investigation revealed that shoot length, plant height and dry weight of
okra plant was maximum in T2 treatment with 18 cm, 26.33 cm and 2.80 g,
respectively, and minimum in chemical fertilizers treated group (T3), while plant
treated with chemical fertilizers showed minimum growth in root length (4.33 cm)
which was comparatively low to vermicompost and control treatment (Table 4.2 &
Figure 4.12). The present research also studied the variable impact of vermicompost
and chemical fertilizers on fruit length and yield (Table 4.3 and Figure 4.13). In this
investigation fruit length was maximum in T2 treatment with 10.6 cm and minimum in
T1 treatment (6.51 cm), maximum fruit width was observed in T2 treatment with 2.63
cm and minimum in T3 group (0.98 cm). Average weight was maximum in T2
treatment with 15.22 gm and minimum in T1 treatment respectively. The highest yield
of okra fruits per plant recorded in T2 treatment. It was 2.83 kg/ plant (Table 4.3).
Table 4.2 Comparative effects of vermicompost and Table 4.3 Comparative effects of vermicompost and
chemical fertilizers on growth of okra plant chemical fertilizers on growth of okra fruits

Treatments Root Shoot Plant Dry Treatme Length Width of Average Quantity
length length Height weight nts of fruits fruits weight (Kg)
(cm) (cm) (cm) (g) (cm) (cm) (g)

T1 9.00 13.33 24.00 1.67 T1 6.51 1.54 4.66 1.5

T2 11.33 18.00 26.33 2.80 T2 10.6 2.63 15.22 2.83

T3 4.33 12.66 18.66 1.45 T3 8.2 0.98 6.09 2.24

Figure 4.12 Growth parameters of okra plant in three various treatments Control (T1), Vermicompost (T2)
and Chemical fertilizers (T3)

T1 T2 T3

Figure 4.13 Effect of control (T1), vermicompost (T2) and chemical fertilizers (T3) on the growth
of okra fruits
T1 T2 T3
The present research also revealed the impact of vermicompost and chemical fertilizers on
nutrition management in okra plant. The micronutrient level, which observed in okra plant
was Mg>Ca>Cu>Mn>Zn (Table 4.4) . The highest Mg, Ca and Cu content of 250, 13.12
and 6.56 mg were recorded with the application of vermicompost. The chemical fertilizer
treatment was recorded the quantity of magnesium, calcium and copper in 98, 11 and 3.28
mg/ g plant extract. In this study, a significant effect of vermicompost on nutrition
management was observed.

Table 4.4 Determination of micronutrients level in okra plant cultivated with three
various treatments (T1) control, (T2) vermicompost and (T3) chemical fertilizers (Urea
& DAP)

Treatments N P K Ca Mg Zn Mn Cu

T1 0.011 16.7 98 8.12 15 3.89 6.71 0.67

T2 0.08 26.0 342 13.12 250 7.95 17.8 6.56

T3 0.06 24.2 170 11.0 98 5.21 9.5 3.28

Eggplant :
The eggplant (Solanum melongena L.) also known as brinjal is a vegetable crop, consumed
widely all around the world. Eggplant contains a variety of protein, vitamins (A, B1, B2,
B3, and C) and minerals such as zinc, magnesium, copper, sulfur, manganese, sodium,
potassium, calcium, phosphorus and iron beneficial for human health (Akanbi et al.,
2007).
The present study conducted to evaluate the effect of organic and inorganic manure on the
growth of eggplant with the reference of shoot length, root length and plant height. Dry
weight of eggplant also observed during the study. In the present study, experiment
designed in three groups control (T1), Soil with vermicompost (T2) and chemical
fertilizers (Urea & DAP) (T3). Table 4.5 and figure 4.14 Shows shoot length between the
range of 9.33cm to 13.66 cm. The maximum shoot and root length (13.66 and 7.75cm)
were recorded in the eggplant cultivated with vermicompost (T2 group), while eggplant
of control and chemical fertilizers did not show significant variation in shoot length and
root length.
The yield of eggplant fruits was found maximum (1.9kg per plant) in vermicompost
treatment. Average weight of fruit also around 26.22 g (Table 4.6 & Figure 4.15). The low
weight of fruits was recorded in control plant.
Eggplant fruits which were grown with vermicompost show a good nutritional
management in both plant and fruit(Table4.7).

Organically grown crops were also found to be superior in their nutritional content along
with growth and yield parameters as compared to chemically grown crops (Jaya & Sunita
2014). Mishra and Katariya, 2018 concluded that vermicompost can be recommended for
better growth and enhancement of phytochemicals in agricultural practices.

Table 4.7 Determination of micronutrients level in eggplant cultivated with three

various treatments (T1) control, (T2) vermicompost and (T3) chemical fertilizers
(Urea & DAP)

Treatments N% P K Ca Mg Zn Mn Cu

T1 0.03 16.3 210 7.25 145 0.41 1.71 1.74

T2 0.04 36.0 360 26.11 370 2.75 6.00 4.95

T3 0.06 44.2 186 8.79 265 1.27 3.25 2.28

PHYTOCHEMICAL TEST
Phytochemical test of eggplant extract

The results of the phytochemical analysis of eggplant extract grown with in three various
treatments show in Table 4.8. The current research confirmed the effect of organic manure on the
accumulation of highly valuable phytochemicals like phenol, flavonoids and alkaloids. Amino acid,
carbohydrate, saponin and other constituents also found in eggplant grown with vermicompost,
while plant cultivated with chemical fertilizers and control did not show the presence of flavonoid
and others, only phenol and carbohydrate observed in the extract of both treatment.
Ibrahim et al., (2013) investigated the comparative effect of organic and inorganic manure on the
phytochemical and antioxidant activity of kacip fatimah (Labisia pumila Benth). They found that
the organic manure enhanced the production of secondary metabolites and improve the antioxidant
activity of this herb.
Table 4.8 Evaluation of phytochemicals in hydroalcoholic extract of eggplant cultivated in
three distinct group (T1) control, (T2) vermicompost and (T3) chemical fertilizers

TEST T1 T2 T3
Carbohydrates + + +
Phenols + + +
Ferric _ _ _

Saponins _ + _

Gums _ _ _
Flavonoids + + -
Alkaloids - + _
Amino acids _ + +
Reducing sugar + + +
Fixed oil _ - -
Table 4.10 total phenol content in hydroalcoholic extract of okra and eggplant grow in control, vermicompost
and chemical fertilizers

T1 T2 T3
Okra 0.054 0.186 0.157
Egg plant 0.20 0.269 0.073

0.3
Total phenol Content (TAE)

0.25

0.2

0.15
Okra
0.1 Egg plant
0.05

0
T1 T2 T3
Treatments

Figure 4.17 total phenol content in hydroalcoholic extract

of okra and eggplant cultivated in three groups
Total Flavonoid Content
The total flavonoid content of hydroalcohol extracts of eggplant and okra cultivated with three
discrete treatments was estimated by aluminium chloride assay. Total flavonoid content expressed as
quercetine equivalent (mg /g QE) in different extracts of both plants has been summarized in Table
4.11. The highest concentration of total flavonoid content was found to be 0.839 mg QE/g in eggplant
extract, which treated with vermicompost, while okra cultivated with organic compost exhibited
0.534 mg QE/g of total flavonoid content (Figure 4.19). Both the plants grow in control treatment did
not show any significant variation in flavonoid content. The eggplants applied with organic sources
of manure , especially vermicompost was exceptionally healthy, taller and heavier plant.
0.9
0.8

Total Flavonoid content (mg/QE)

0.7
T1 T2 T3 0.6
0.5

Okra 0.496 0.534 0.368 0.4

Okra
0.3 Egg plant
0.2
Egg plant 0.419 0.839 0.673 0.1
0
T1 T2 T3
Treatments
Table 4.11 Total flavonoid content in
Table 4.19 total flavonoid content in hydroalcoholic
hydroalcoholic extract of okra and eggplant
extract of okra and eggplant cultivated in three groups
grown in control, vermicompost and
chemical fertilizers
ANTIOXIDANT ASSAY
DPPH Assay
The antioxidative potential of the hydroalcoholic extract of the okra and eggplants grown
in three different soil treatments i.e., untreated natural soil (T1), soil treated with
chemical fertilizers (T3) and soil treated with vermicompost (T2), was observed by
measuring changes in absorbance of DPPH radical at 517 nm because of their radical
scavenging effect. The scavenging activity against DPPH free radical has been
extensively used to evaluate the antioxidant activity of plant extracts. In the present
study maximum percentage of scavenging was recorded vermicompost treated plants
(Figure 4.21). Comparatively eggplant cultivated with vermicompost scaveng highest
49.25% DPPH radicals while okra extract reduces 25.37% DPPH.

100
60
90
80 50
% of Scavenging

70 % of Scavenging 40
60
50 30
40 Okra
20
30 Eggplant
20 10
10
0
0
T1 T2 T3
0 0.2 0.4 0.6 0.8 1 1.2
Treatment
Concentration (Mg)

Figure 4.21 DPPH assay of okra and eggplant extract

Table 4.20 Standard graph of L- ascorbic acid for DPPH assay cultivated with inorganic and organic manure
Nitric Oxide Radical Scavenging Activity

Figure 4.22 shows the free radical scavenging activity of okra and eggplant cultivated
with three various treatments (T1) control, (T2) vermicompost and (T3) chemical
fertilizers (urea & DAP). It shows that extract of both plants cultivated with
vermicompost has the property of scavenging nitric oxide radicals. The nitric oxide
radical scavenging of okra plant treated with vermicompost was 65.67%, which was
higher than NO scavenging activity of eggplant (52.23%), but the chemical fertilizers
treated plants of okra and eggplant showed lowest radical scavenging activity.
70

60

50
% of NO scavenging

40

30 Okra
Eggplant
20

10

0
T1 T2 T3
Treatments

Figure 4.22 Nitric oxide scavenging assay of okra and eggplant extract cultivated with inorganic and organic manure
Total Antioxidant activity

The results of the total antioxidant activity of okra and eggplant extracts are demonstrated in
Figure 4.23. Among all the treatments, vermicompost treated plants of okra and eggplants
showed highest total antioxidant activity. However, total antioxidant capacity of the control
plants was significantly lower. Okra plants cultivated with vermicompost reduced 65.06%
molybdate ions which was higher in comparison of eggplant extract (63.85%). Whenever
control and chemical treated eggplant extract showed more activity than okra (Figure 4.23).

Boubekri et al., (2015) reported that the organic compost enhance the level of phenol in the
various parts of plants and phenolic compounds have a significant contribution to the total
antioxidant activity.
70

60

50
% of Scavenging

40

30 Okra
Eggplant
20

10

0
T1 T2 T3
Treatments

Figure 4.23 Total antioxidant assay of okra and eggplant extract cultivated with inorganic and organic manure
 SUMMARY AND CONCLUSION

Vermicompost is the product of the composting process using various species

earthworms. vermicompost can enhance soil fertility physically, chemically and
biologically. Physically, vermicompost-treated soil has better aeration, porosity, bulk
density and water retention. Chemical properties such as pH, electrical conductivity
and organic matter content are also improved for better crop yield.
Vermicompost contains water-soluble nutrients and is an excellent, nutrient-
rich organic fertilizer and soil conditioner.
The present research entitled Study the effect of chemical and organic manure in
micronutrients of vegetables. The thesis encloses the original work carried out at
Department of Chemistry, Govt. Gitanjali Girls P.G. College, Bhopal.
The present study aims to develop economical organic manure like vermicompost
which enhance the nutritional values of plants and manage the level of
micronutrients.
The present work was commenced to fulfill the following objectives:

• To assess the potential of the earthworm species (Eisenia foetida) to transform

agricultural waste into useful compost.

• To evaluate the physicochemical parameters of the soil.

• To evaluate the physicochemical parameters of the vermicompost produced.

• To evaluate the effect of organic manure (vermicompost) on vegetable crops as

compare to chemical fertilizers.

• To evaluate the phytochemical and antioxidant activity of vegetable crops.

The current investigation revealed that shoot length, plant height and dry weight of okra
plant was maximum in T2 treatment with 18 cm, 26.33 cm and 2.80 g, respectively, and
minimum in chemical fertilizers treated group (T3), while plant treated with chemical
fertilizers showed minimum growth in root length (4.33 cm) which was comparatively low
to vermicompost and control treatment (Table 4.2 & Figure 4.12). The present research
also studied the variable impact of vermicompost and chemical fertilizers on fruit length
and yield (Table 4.3 and Figure 4.13). In this investigation fruit length was maximum in
T2 treatment with 10.6 cm and minimum in T1 treatment (6.51 cm), maximum fruit width
was observed in T2 treatment with 2.63 cm and minimum in T3 group (0.98 cm).
The present research also revealed the impact of vermicompost and inorganic fertilizers on
nutrition management in okra plant. According to the current results, okra plants treated
with vermicompost showed higher concentrations of macro nutrients (nitrogen,
phosphorus and potash).The chemical fertilizer treatment was recorded the quantity of
magnesium, calcium and copper in 98, 11 and 3.28 mg/ g plant extract. In this study, a
significant effect of vermicompost on nutrition management was observed.
Table 4.5 and figure 4.14 Shows shoot length between the range of 9.33cm to 13.66 cm.
The maximum shoot and root length (13.66 and 7.75cm) were recorded in the eggplant
cultivated with vermicompost, while eggplant of control and chemical fertilizers did not
show significant variation in shoot length and root length.
Total flavonoid content expressed as quercetine equivalent (mg /g QE) in different
extracts of both plants has been summarized in Table 4.11. The highest concentration of
total flavonoid content was found to be 0.839 mg QE/g in eggplant extract, which treated
with vermicompost, while okra cultivated with organic compost exhibited 0.534 mg QE/g
of total flavonoid content (Figure 4.19).
In the present study maximum percentage of scavenging was recorded vermicompost
treated plants (Figure 4.21). Comparatively eggplant cultivated with vermicompost
scaveng highest 49.25% DPPH radicals while okra extract reduces 25.37% DPPH.
Figure 4.22 shows the free radical scavenging activity of okra and eggplant cultivated
with three various treatments (T1) control, (T2) vermicompost and (T3) chemical
fertilizers (urea & DAP). It shows that extract of both plants cultivated with
vermicompost has the property of scavenging nitric oxide radicals. The nitric oxide
radical scavenging of okra plant treated with vermicompost was 65.67%, which was
higher than NO scavenging activity of eggplant (52.23%), but the chemical fertilizers
treated plants of okra and eggplant showed lowest radical scavenging activity.
The results of the total antioxidant activity of okra and eggplant extracts are demonstrated
in Figure 4.23. Among all the treatments, vermicompost treated plants of okra and
eggplants showed highest total antioxidant activity. However, total antioxidant capacity of
the control plants was significantly lower.