CONVERSION OF DIFFERENT RAW MATERIALS INTOCOMPOST BY EFFECTIVE MICROORGANISM (EM) ANDTHEIR EFFECTS ON POST HARVEST QUALITY OF

CHINESE SPINACH (Amaranthus oleraceus)

A proposal by

NURUL FAIQAH BINTI MOHD NOR

Supervisor

Puan Ramisah binti Mohd Shah

Co-supervisor

Dr Wan Zaliha binti Wan Sembok

SCHOOL OF FOOD SCIENCE AND TECHNOLOGY

UNIVERSITY MALAYSIA TERENGGANU2014/2015

CHAPTER 1

INTRODUCTION

1.1 Background of study

Chinese spinach (Amaranthus oleraceus) mainly originated from

African, American, Asian tropics. The consumption of this kind of

spinach has been established for food uses in India and Nepal.

Chinese spinach is classified under family Amaranthaceae. It is

extensively grown as a green, leafy vegetable in humid tropics

because of its high advantages such as able to withstand the hot

weather environment, high yields and contain high nutritive

value. On the other hand, spinach is an annuals crop. In the

previous study, according to its high nutritive value as well as

the ability to adapt to drought stress, vegetable amaranth is a

promising C4 crop especially for the semi-arid environment (Liu

and Stutzel, 2004). Amaranthus oleraceus has high potential and

economic value crop yield.

Nowadays, the application of agrochemicals in agriculture

industry has polluted the environment. Furthermore, the food

produced by the farms with the application of the agrochemical

may not be safe or of good quality. Public awareness to these

problems resulted in some approaches towards the discoveries of

some alternative measures (Shaxson, 2006). Far and foremost, the

implementation of the organic farming offers an alternative that

can eliminate a lot of the environmental problems of conventional

agriculture. In contrary to the conventional farming, organic

farming developed potential benefits in improving food safety and

quality (Giles, 2004), promoting the formation of the soil

structure (Pulleman et. al., 2003), alleviating environmental

stresses (Macilwain, 2004), as well as enhancing soil

biodiversity (Oehl et., al., 2004).

The sustainability of plant production is related to soil

fertility. Improving soil fertility therefore contributes to a

sustainable production of healthy plants. Effective Microorganism

(EM) was originated from the Asia country that was in Japan.

Nowadays, EM is widely applied in sustainable and organic

farming. This product was pioneered by T. Higa from Ryukyus

University, Okinawa, in the early 1980s. The main microorganism

in the EM are lactic acid bacteria (Lactobacillus plantarum L. casei,

Streptoccus lactis), yeast (Saccharomyces cerevisiae, Candida utilis),

photosynthetic bacteria (Rhodopseudomonas palustrus, Rhodobacter

spaeroides), actinomycetes (Streptomyces albus, S. griseus) and fungi

(Aspergillus oryzae, Mucor hiemalis) (Daly & Steward 1999; Higa, 2002).

EM fermentation can be composed in two ways that are the aerobic

and anaerobic fermentation. The mixture was sold as the

stimulator of substrate mineralization and plant growth (Higa &

Parr, 1994) and also applied as “bokashi”. Bokashi is the results

of a fermentation of organic substances, for instance, the wheat

bran, with addition of EM. Applying EM is not a substitute for

other agricultural starategies but rather an additional dimension

to optimize the practices and productivity in organic farming

(Higa, 2004).

Pioneered by the research done by Higa in Japan (Higa,

2004), the potential of EM has been studied in many countries

around the world. As a result, some countries indicated or showed

positive effects on the application of the EM onto the soils and

plants on the quality and nutrient delivery of the soils

(Aylesworth, 1979; Xu, 2001), on plant growth (Aylesworth, 1979),

on crop yield (Xu, 2001), and on crop quality (Higa, 2004).

1.2 Problem statement

Nowadays, the agriculture sectors have contributed to the

pollution due to the implementations of agrochemicals in our

daily local agriculture environments. If this problem continues

to rise, it will generate more severe effects to our future

environments. Hence, the implementation of the sustainable

agriculture is fundamental in order to prevent the problem.

Sustainable agriculture practices can be instilled by using the

organic farming method instead of the chemical uses. Chemical

substance in agricultural practices will disrupted the mineral of

the soil as well as will contaminate the underground water

sources in a long term effects. Another alternative method that

can be taken to replace the chemical used especially in the

fertigation system is by the application of the organic compost.

For instance, the organic compost derived from the bokashi and

coir pitch fermentation can be easily done. By this research, the

further progress of the implementation of organic compost from

the raw materials of bokashi and coir pitch as well as the

addition of the effective microorganism can be proven to produce

a high quality of crops.

1.3 Significance of study

The purpose of this research is to find out some

natural ways for minimizing the dependency on agrochemical

fertilizer in order to increase the production of yield and

improve crop quality of Amaranthus oleraceus.

Natural way of agricultural practice that will be

conducted in this research is by applying the different raw

materials as compost which acts as organic fertilizer in

combination with the effective microorganism. The raw materials

used in this research are the bokashi and the coir pitch. In

Malaysia, coir pitch and all the raw materials needed to ferment

the bokashi are readily available in no time. It also does not

require much cost in order to get the raw materials.

In addition, by using the crop residue such as the coir

pitch and the rice bran can help to clean the environment and

better agricultural sustainability by minimizing waste as well as

recycling by-products. This can improve agricultural

sustainability, save cost and environment friendly by reducing

the dependence on agrochemicals.

1.4 Objectives

1. To study the effects of effective microorganism (EM) by

different types of raw materials of composts to the growth

of the Chinese spinach (Amaranthus oleraceus).

2. To observe the physiochemical interaction between the

effective microorganism (EM) to the application of composts

as well as morphological effect in plant growth.

3. To find the best collaboration between the effective

microorganism (EM) and the raw materials of the compost.

CHAPTER 2

LITERATURE REVIEW

2.1 Chinese Spinach (Amaranthus Oleraceus)

Chinese spinach (Amaranthus oleraceus) has become an

important vegetable crop in most regions of the world and

remarkable changes in production amount have occurred in the past

decades due to demand increase in many countries. Spinach

(Spinacia oleracea L.) is an edible and annual plant that grows

rapidly and has the ability to survive over moderate winter. It

is versatile which is used as a salad, a cooked vegetable or as a

component of many other cooked meat and vegetable dishes (Sensoy

et al., 2011). Leafy vegetables are an important part in the

human diet and spinach is one of the dark green leafy vegetables

which contains high beta carotene and folate, and is also a good

source of vitamin C, calcium, iron phosphorous, sodium and

potassium (Dicoteau, 2000; Avsar, 2011). Spinach as dioecious

species with both male and female plants is an herbaceous leafy

vegetable in the family of Amaranthaceae (Salk et al., 2008) and

its leaves are alternate, simple, from ovate to triangularbased,

with larger leaves at the base of the plant and small leaves

higher on the flowering stem (Vural et al., 2000). In a previous

study, Amaranthus oleraceus are grouped of under utilize or rather

exploited crops comprising of grain, leaf, forage and ornamental

types (Yadav and Rai, 2005). Amaranth has become a focus of

nutritional and agronomic research and it is on the verge of

commercial exploitation over the past decade (Tucker, 1986).

Varieties of this species are native from a large area from India

to the island of the Pacific and as far north as China. It is

probably the best developed of the vegetable amaranth species.

They can be produced as a hot-season leafy vegetable in arid

regions when few other leafy greens are available (Yadav and Rai,

2005).

Today, China, the United States, Indonesia, Japan and Turkey are

among the largest commercial producers of spinach (FAO, 2011).

Iran is the one of the spinach producers with about 105 thousand

tons per year based on FAO statistics. The average yield of

spinach in Iran is 2096 kg ha-1 while world’s average yield is

2420 kg ha-1 (FAO, 2011). Also, the average yield of spinach in

China is 2768 kg ha-1, in the United States is 2360 kg ha-1,

Indonesia is 3424 kg ha-1, Japan is 12471 kg ha-1, and Turkey is

9249 kg ha-1 (FAO, 2011). Spinach is native to southwest Asia and

commonly thought to have originated in Iran (Nonnicke, 1989;

Swiader and Ware, 2002) and was first mentioned by the Chinese as

the herb of Persia. It was first cultivated in North Africa, came

to northern Europe by way of Spain, documented in Germany and

then was a common garden vegetable by 1500 in England and France

(Dicoteau, 2000; Swiader and Ware, 2002).

Figure 2.1 Chinese spinach (Amaranthus oleraceus)

2.2 Effective Microorganism (EM)

The concept of Effective Microorganism (EM) was developed in

1971 by Professor Teruo Higa, university of the Ryukyus in

Okinawa, Japan. EM solution is prepared from natural substrates

and it can be used as an herbal insecticide to control insects

and pathogenic organisms and can also used as plant growth

inducers. The use of EM in crop production is either by direct

application to the soil composition, then the the action of the

organic matter started as it spread to the plant or even to the

surface of the soil. However, it can be applied to the soil prior

planting (Shintani, 1995). Studies have suggested that EM may

have a number of methods in implementation, including

agriculture, gardening, livestock, landscaping, sanitization of

the septic tanks, composting, bioremedication, algal control as

well as for the house hold applications. The present study was

undertaken to investigate the effects of the effective

microorganism (EM) by different types of raw materials of

composts in tomato cultivation. Raw materials that are included

in this research are comprising of Bokashi and coir pitch.

2.3 Compost

Composting is the biological process of decomposition of organic

constituents of bio-waste materials under controlled conditions

(Golueke, 1972; Hoitink and Keener, 1993). During this process,

carbon from organic molecules gets converted to carbon dioxide,

resulting in the reduction of bulkiness of the organic forms,

which can be absorbed directly by plants. Rates of decomposition

and mineralization of organic residues differ among species

having different plant chemistry (Palm and Sanchez, 1991 ).

Composting rate depends to a great extent on C:N ratio, lignin

and polyphenol contents, presence or absence of suitable

microbial agents of decomposition such as the lignin. Commonly,

compost is an organic matter that has been decomposed and

recycled as fertilizer and soil amendment. The abundance of raw

materials as organic waste can be converted into fertilizer

throughout the process of composting.

Composting is a biological treatment that is cost-effective to

treat different types of organic waste (Tiqua and Tam, 2002).

Composting is the first concept for using effective microorganism

(EM) in environmental management. Crop residues and animal wastes

have been effectively composted to produce biofertilzers.

2.4 Bokashi

Bokashi means fermented organic matter in Japanese. Bokashi

composting uses a selected group of microorganisms to

anaerobically ferment organic waste. The microorganisms are

applied using an impregnation carrier such as wheat bran. The

fermentation process breaks the organic matter down in a process

that is odor free. Bokashi is produced by adding EM-2 to a high

carbon substrate, originally rice bran, but sawdust is now

commonly used, often along with some additional nutrient sources,

such as molasses, and then fermenting it. This provides a stable,

easy to handle material. This can be done under quite low-tech

conditions, e.g., by farmers in barns, but for a more consistent

material commercially produced Bokashi is likely to be best. Most

fermentation processes require inoculation with suitable microbes

to ensure fermentation occurs as required. Most fermentation

processes take often relatively homogeneous aseptic starting

material while food ‘waste’ by its nature is heterogeneous and

will be pre-colonised by a diverse range of microbes, including

many decomposers. Inoculating such material with suitable

fermentation microbes is therefore a not an insignificant task,

especially as one species on its own may not be enough. One of

the interesting aspects of EM is the general stability of the

cultures and their consistency, even though where samples of EM

are taken at point of use and analysed for the presence of the

microbes that went into EM, not all of the types of microbes are

present (Yamada & Xu, 2000). Therefore the reports from the

literature demonstrating the ability of EM to consistently

ferment materials, such as food ‘wastes’, is both impressive and

consistent with general experience with EM. Also there are a few

reports looking at fermenting food ‘waste’ using the naturally

occurring microbes or with single species cultures, none of which

fermented as well as when Bokashi was used (Yamada & Xu, 2000).

This is also consistent with theory. Bokashi therefore appears to

be very well suited as the starter culture for fermenting food

preparation ‘waste’ and it ready availability and comparatively

low cost means there is little value in considering other starter

cultures, at least initially or if problems are found.

2.5 Coir pitch

India currently produces over 15,000 million coconuts per year.

In addition to the utilization of

endosperm for edible purposes and extraction of oil, the outer

non-edible fibrous portion of the nuts (coconut husk) is used for

extracting coconut. fibre or coir, which is commerciality

utilized for making value-added products such as mats,

geotextiles etc. In the husk. coconut fibres are seen tightly

packed along with non-fibrous, fluffy and light weight croaky

material known as coir pith or coir dust, which constitutes about

50-70 percent of the husk. The composition and properties of coir

pith vary (Moorthy and Rao, 1998) depending on maturity of

coconut, method of extraction and disposal,period between

extraction and use and environmental factors. Wide variations in

C:N ratio of coir pith from 58:1 to ll2: 1 has been reported

(Savithri and Khan, 1994). Retted husks yield coir pith with less

nutrients than that obtained by mechanical processing of unretted

husk. Coir pith obtained from fully mature nuts has higher

amounts of lignin and cellulose and Jesser amount of water

soluble salts compared to younger nuts. When husks of 10,000

coconuts are utilized for coir extraction, one tonne of coir pith

is obtained as a by-product. If all the coconut husks available

in lndia are processed, it is estimated that about 1.5 million

tonnes of coir pith could be obtained anually. But in reality,

all the available coconut husks are not diverted for coir

extraction and it has been reported that only 5 lakh tonnes of

coir pith is produced in India annually (Joseph, 1995). Because

of high fertilizer prices and environmental concerns associated

with its use and with the enhanced emphasis on commercial

horticulture and organic farming, recent years have witnessed

growing interest in utilizing coir pith in a more productive way

in agrihorticulture (Pmbhu and Thomas, 2001). Coir pith has got

many enviable characteristics making it a highly potential

resource if used after proper composting. Coir pith has very high

moisture retention capacity of 500-600 per cent and can be as

high as 1100% of dry weight (Evans er al., 1996). In a

comparative study with coir pith and three types of saw

dust, coir pith retained the maximum moisture after 90 days

of composting (MlY.lh and Pdili, 1998). It has high

potassium content and low bulk density and particle density.

The low particle density is due to high specific surface and

high specitic surface gives it high cation exchange capacity

(CEC) (Mapa.and Kumara, 1995). High CEC, which varies from

38.9-60 meq/lOOg (Evans e.t aL, 1996) enables it to retain

large amounts of nutrients and the adsorption complex. bas

high contents of exchangeable K. Na, Ca and Mg (Verhagen and

Papadopoulos, 1997). All these characteristics make it ideal

for use as a mulch and soil amendment, especially for dry

and sandy areas with low water retention. The stabilized,

compostcd coir pith resembles peat and has got many

characteristics as that of sphagnum peat. The most common

potting media used in horticulture and hence it is

commercially known as coco peat. With the development of

commercial horticulture and reduction in the availability of

sphagnum peat, coco peat has become internationally

recognised as an ideal soil amendment and component of soil-

less container-media for horticulture plants. Coco peat find

use in germination of seeds, nursery raising, rooting of

cutting and other vegetative plant propagation methods,

hardening of tissue and embryo cultured plants, hydroponic

systems of plant cultivation, cultivation of glass house

plants, soil conditioning, lawn making etc. (Bavappa and

Gurusinghe, 1978, Ani.tha Karon et al., 1999, Rao, 1999).

CHAPTER 3

METHODOLOGY

3.1 Location of experiment

The location will be conducted at the glasshouse of

University Malaysia Terengganu.

3.2 Materials

Amaranthus oleraceus will be supplied by …… Coir pitch will

be harvested from the coconut plantation around Terengganu

areas. Wheat bran and fish meal that will be used in making

the Bokashi compost will be supplied by …..

3.3 Composting

3.3.1 Composting of coir pitch

Coir pith is collected from the coir industry without any

fiber. If fibrous materials are present, it is removed by

sieving at the source itself. Otherwise, it has to be

removed at the end of composting at the compost yard. These

fibrous materials will not get composted and it will hinder

with composting process. It is advisable to bring fibre free

coir pith for composting. A separate area should be

earmarked for composting. It is better to have an elevated

place for composting. Coir pith is an aerobic composting. So

it should be heaped above the soil. There is no need for pit

or cement tub to make the compost. After moistening,

nitrogenous source material should be added. The nitrogenous

source may be in the form of urea or fresh poultry litter.

If urea is applied, it is recommended that 5 kg urea is

required for one ton of coir pith. Thus for this research, 5

ml of urea is needed to compost 3 kg of coir pitch. The

compost heap should be turned once in 10 days to allow the

stale air trapped inside the compost material to go out and

fresh air will get in. The composting process is an aerobic

one, the organism performing the composting require oxygen

for its metabolic activity. This turning of material

indirectly aerate the substrate. The other way of giving

aeration is inserting perforated unused PVC or iron pipe in

the composting material both vertically and horizontally.

Maintaining optimum moisture is the pre-requiste for uniform

composting or waste material. Sixty percent moisture is to

be maintained 60 % moisture is, the compost material should

be always wet. But excess water should not be drained form

the waste material is to take a handful of composting

material and put in between the palms and squeeze it. If no

water is coming out of the material, that moisture status is

ideal for composting. The period of composting vary from

substrate to substrate. If all the above said conditions are

maintained in the composting, it will take sixty days (60

days) for some of the physical parameters to be observed in

the compost. 

3.3.2 Composting of Bokashi with EM-2

2 litre of the water is mixed with 3 kg of rice bran along

with the mix oil cake and the fish meal. 60 ml of EM-2 is

added. Lightly squezze some of the rice bran into ball shape

to check the moisture level. If it holds shape and no extra

liquid comes out, it is the correct moisture. If it is too

dry, add more water and mix. The mixture is put into the

airtight container, press down the mixture. If using the

bag, press it tightly, squeezing out, excess air. If using

container, press down the mixture to let it absorb the extra

moisture and cover container tightly. Place the mixture

somewhere warm and keep out of direct sunlight. Let it

ferment for a month. When the colour turns darker and it

smells sweetened-and-sour, fermentation is completed. To

keep it in the airtight container or let it dry in the shade

enables the longterm storage. The same method of bokashi

fermentation without the EM-2 can be done without adding 60

ml of EM-2.

3.4 Experimental Design

The raw materials that are the ‘Bokashi’ and the

coir pitch in combination of the effective microorganism

(EM) are used as the compost making in the following

treatment.

Treatment 1: 3 kg of Bokashi

Treatment 2: 3 kg of Bokashi + 60 ml of EM-2 solution

Treatment 3: 3 kg of coir pitch

Treatment 4: 3 kg of coir pitch + 60 ml of EM-2 solution

The composting is done by sandwich method in a pit (45

cm height and 30 x 20 cm in diameters) or the Bokashi is

fermented in a container. The pits anf the container are

covered with polyethylene sheet. Moisture content is

maintained during composting by spraying water regularly.

The compost will be ready after 45 days.

RCBD Field Layout

T1R2

T1R4

T2R3

T2R1

T2R4

T2R5

T2R

T4R4

T4R3

T4R2

T4R1

T4R5

T3R1

T3R2

T3R3

T3R5

T3R4

3.5 Compost analysis

The representive compost samples were collected and

air-dried. The following physical and chemical properties of

the compost were analyzed.

3.5.1 Total nitrogen

The total nitrogen will be measured by using the Nitrogen

Alamantar device.

3.5.2 Cellulose

The total cellulose will be measured by using the Fiber

Analyser device.

3.5.3 рН

The pH will be measured by using the pH meter.

3.6 Pot Cultivation

Amaranthus oleraceus are germinated in 35 x 25 cm trays. 20

polybegs (diameter 16.5 cm, height 14.5 cm) are prepared in

5 replications for each prepared treatment of the compost

(Bokashi and coir pitch). Then each of the polybeg is filled

with the briss soil (2 kg) in exact amount as the culture

medium. About 5 of A. oleraceus seedlings at the stage 3 to 4

were transplanted in each of the pot. All treatments were

arranged in completely randomized design with 4 replicates.

The pots are provided with water facilities. The pots are

maintained in the open shade at the temperature of 27-30°Ϲ.

3.7 Analysis of morphological parameters

After 4 weeks of growth, the plants are removed

from all the samples and studied for the following

morphological parameters such as the height of the plant (in

cm), number of leaves (per plant), shoot fresh weight (per

plant), the number of roots (per plant) and the chlorophyll

a, b, and caretonoids.

3.7.1 Height of plant

Height of plant was measured by using the metre rule.

3.7.2 Number of leaves

Number of leaves were counted manually and recorded.

3.7.3 Shoot fresh weight

Shoot fresh weight was weighted by using electronic balance

and the emergence is counted manually.

3.7.4 Total leaf area

Leaf area was measured by using leaf meter area.

3.7.5 Chlorophyll a, b, and carotenoids

750 mg of A. oleraceus were weighted. Then, 1 g

of magnesium oxide and 25 ml acetone were added. The samples

were placed in mortars and were ground with pestle. The

extracts were transferred into 50 ml falcon centrifuge tube.

The extracts were rinsing with a further 15 ml of 100%

acetone. The extracts were centrifuge at 2000 rpm for 5

minutes. The supernatant were decanted into 50 ml volumetric

flask and make up to volume using 100% acetone.

The absorption of chlorophyll a, b, and

carotenoids was measured by using UV-vis spectrophotometer

at wavelength 750, 662, 645, 520, and 470 nm.

3.8 Statistical Analysis

Mean was calculated to facilitate the comparison

of the data of various physiochemical parameters of compost

and growth parameters of green grams in all samples.

CHAPTER 4

EXPECTED RESULT

The effect of Effective Microorganism (EM) on the

compost with different raw materials that are the bokashi as

well as the coir pitch was reflected on the total nitrogen,

cellulose content and the pH measurement of physiochemical

parameters. The compost was ready after 45 days. After

composting, the effect of compost on the growth of Chinese

spinach (Amaranthus oleraceus) was reflected on the height of

plant, number of leaf, shoot fresh weight, total leaf area

and also chlorophyll a, b, and carotenoids.