The Viability of African Nightcrawlers (Eudrilus eugeniae) as a Protein Source for

Broiler Chicken

NICOLE DOMINIQUE G. DELA CRUZ

ANA MARLA M. NAWE

Researchers
 ABSTRACT

Livestock production is the agricultural activity with the greatest environmental

impact. Animal agriculture frequently has a negative impact on the environment that is

mostly caused by the feeding of livestock. Consuming protein-rich feed is essential for

attaining sustainable production methods, especially in intense production. This study

investigated the viability of African nightcrawlers (Eudrilus eugeniae) as a protein source

for broiler chicken in comparison to commercialized isolate soy protein in terms of their

physical properties, chemical composition, nitrogen free extract (NFE) and metabolizable

energy (ME). 200 grams of each sample were sent to the Region 3 Feed Chemical

Analysis Laboratory. Nitrogen free extract (NFE) and metabolizable energy (ME) were

tested by this testing center. The physical properties of African nightcrawlers (Eudrilus

eugeniae) were found to be appropriate for broiler chicken feeding. However, the

chemical composition of African nightcrawlers (Eudrilus eugeniae) were not found to be

completely suitable for broiler feeding. Furthermore, the results lead to the acceptance of

the null hypothesis that there are no significant differences between African nightcrawlers

(Eudrilus eugeniae) and commercialized isolate soy protein in terms of NFE. At the same

time, it is also concluded that there are significant differences between the two groups in

terms of ME. Given the results, it may be concluded that this study not only shed light to

the promising potential of vermicomposting worms, specifically African nightcrawlers

(Eudrilus eugeniae), as a protein source for poultry feeding but also in unlocking new

knowledge that offers insights and builds foundation for future research, practice, or

policy.

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 ACKNOWLEDGEMENT

First and foremost, we would like to praise and thank God, our Almighty, for

showering us with strength and wisdom throughout our research work to complete the

study successfully.

We would also like to express our deepest and sincerest gratitude to our research

teacher, Ma’am Cristie Ann G. Mauricio, for giving us the opportunity to do research.

Your sincerity, motivation, consideration, and vision have deeply inspired us. It was a

great privilege and honor to work and study under your supervision.

Additionally, we are extremely grateful to our parents for their undying love and

support. Lastly, we would also like to express our heartfelt gratitude to the Region 3 Feed

Chemical Analysis Laboratory, especially to Miss Joyce and Abigail Beltran who

accommodate us. We are very blessed that we had you all throughout our research work.

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 TABLE OF CONTENTS

ABSTRACT………………………………………....……………………………. 01

ACKNOWLEDGEMENT …………………………………….………………… 02

CHAPTER I: THE PROBLEM AND ITS BACKGROUND ……………………. 05

Introduction …………………………………………….………………… 05

Statement of the Problem …………………………….………………...… 06

Hypothesis ………..……………………………………………………… 08

Significance of the Study ……………………….………………………… 05

Scope and Delimitation ……..……………………………………………. 10

Definition of Terms ………………………………………………………. 10

CHAPTER II: REVIEW OF RELATED LITERATURE AND STUDY …….…. 13

Related Literature ……………………………….….……………………. 13

Foreign ………………….……………….….……………………. 13

Related Studies …………………………………………………………… 16

Foreign ………………….……….….……………………………. 16

Conceptual Framework ……………………………..……………………. 18

CHAPTER III: METHODOLOGY ……………..……...…..……………………. 20

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 Research Design ……………………………...…..………………………. 20

Procedure ..………………………………..…….………………………… 20

Data Gathering …………………………….……………………...………. 22

Statistical Treatment ……………………………...………………………. 27

Risk and Safety …………………………….……………………..………. 27

CHAPTER IV: RESULTS AND DISCUSSION ….……………………..………. 28

Discussion ………………………………..…….………………………… 34

CHAPTER V: CONCLUSION AND RECOMMENDATIONS ………..………. 37

Conclusion ………………………………..…….………………………… 38

Recommendations ………………………..…….………………………… 39

APPENDICES ……………………………………..……………………………. 41

REFERENCES ……………………………………..……………………………. 42

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 CHAPTER I

THE PROBLEM AND ITS BACKGROUND

Introduction

Animal protein is currently becoming a more valued resource in the food and

agriculture industries. A variety of investigations are being conducted to find alternate

protein sources. The usage of earthworm biomass is one of the most promising.

Lawrence and Millar (1945) were the first to focus on vermiculture as a source of

extra protein (Lumbricus rubellus). Per dry weight, earthworms have 62.0–71.5 percent

total animal protein and 1.5 percent total fat. In addition, earthworm protein contains "all

amino acids acquired through the breakdown of animal tissues."

Earthworms (Eisenia fetida, Eisenia andrei, Perionyx excavatus, Eudrilus

eugeniae, and Dendrobaena veneta) can be used not only for organic waste recycling but

also as a renewable supply of high-quality fodder premixes, according to research

undertaken in the United States and the United Kingdom.

McInroy's first scientific work, published in 1971, established the groundwork for

several chemical and biochemical studies of various earthworm tissue components in

nations such as the United States, the United Kingdom, Germany, China, the Philippines,

Japan, India, and others. The methods for obtaining dry and paste protein samples were

established, and experiments were conducted to see if they could be used as mixed feed.

Furthermore, among the various agricultural activities, livestock is one of the most

environmentally damaging. Livestock feeding is frequently the primary source of

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negative environmental effects associated with animal agriculture. Protein feed

consumption is a critical component in achieving sustainable production processes,

particularly in intensive production. Because of the associated environmental impact and

rising pricing, real usage of soybean meal and fish meal is not sustainable.

Lastly, it is on this premise that the researchers are prompted to investigate the

viability of African nightcrawlers (Eudrilus eugeniae) as a protein source for broiler

chicken.

Statement of the Problem

This research will focus on the viability of African nightcrawlers (Eudrilus

eugeniae) as a protein source for broiler chicken. Furthermore, Furthermore, it sought to

answer the following questions:

1. How does the African nightcrawlers (Eudrilus eugeniae) be described as a protein

source for broiler chicken in terms of its:

1.1 Physical Properties

1.1.1 Ash Content

1.1.2 Moisture Content

1.1.3 Dry Matter

1.2 Chemical Composition

1.2.1 Crude Fat

1.2.2 Crude Fiber

1.2.3 Crude Protein

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 2. How does the commercialized isolate soy protein be described as a protein source

for broiler chicken in terms of its:

2.1 Physical Properties

2.1.1 Ash Content

2.1.2 Moisture Content

2.1.3 Dry Matter

2.2 Chemical Composition

2.2.1 Crude Fat

2.2.2 Crude Fiber

2.2.3 Crude Protein

3. Is there a significant difference between the viability of African nightcrawlers

(Eudrilus eugeniae) and commercialized isolate soy protein as a protein source for broiler

chicken in terms of:

3.1 Nitrogen Free Extract (NFE)

3.1.1 Ash Content

3.1.2 Moisture Content

3.1.3 Crude Fat

3.1.4 Crude Fiber

3.1.5 Crude Protein

3.2 Metabolizable Energy (ME)

3.2.1 Gross Energy (GE)

3.2.2 Nitrogen Free Extract (NFE)

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 3.2.3 Crude Fat

3.2.4 Crude Fiber

3.2.5 Crude Protein

Hypotheses

There are no significant differences between the viability of African nightcrawlers

(Eudrilus eugeniae) and commercialized isolate soy protein as a protein source for broiler

chicken in terms of nitrogen free extract (NFE).

There are no significant differences between the viability of African nightcrawlers

(Eudrilus eugeniae) and commercialized isolate soy protein as a protein source for broiler

chicken in terms of metabolizable energy (ME).

Significance of the Study

This study investigating the viability of African nightcrawlers (Eudrilus eugeniae)

as a protein source for broiler chickens benefits the following individuals: poultry

owners, farmers, and consumers, to our environment, and as well as to the researchers of

the upcoming generations.

As the aim of this study is to produce an alternative protein source for broiler

chickens which is sustainable and effective thru vermicomposting worms, poultry owners

are especially advantaged. Most commercial protein sources are not sustainable and

costly. However, vermicomposting worms are cost-effective as they can consume litters

and do not need any other supplements other than their feed.

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 Crop residues and agricultural byproducts like husks, bagasse, straw, are often a

problem disposing for farmers. Most farmers resort to burning them, which might be bad

not just for the environment, but also for their health. Improper disposal of animal

manure can also be detrimental to our environment, causing bacterial infection and

contamination to water forms. Through vermicomposting as one of this study’s methods,

these residues and animal dung can be put into use as feed for the earthworms.

The food consumption survey of the Food and Nutrition Research Institute of the

Department of Science and Technology (FNRI-DOST) in 2008 concluded that eggs are

the most consumed protein-rich food in the Philippines. Producing quality eggs benefits

the consumers by providing them the nutrients they need without the fear of contaminants

like heavy metals.

This study uses earthworms that consume organic materials and transform them

into humus-like material called vermicompost that can be used as fertilizer. This

vermicompost plays a major role in the soil structure as they induce soil movement,

nutrient flow, water movement, and plant growth. These earthworms also help manage

wastes as they break down organic materials.

Lastly, this could serve as the foundation to our upcoming researchers in

improving and further utilization of vermicomposting worms in order to unlock new

innovations, alternatives, and solutions that aim to solve crises, and improve living whilst

still putting in mind the environment’s safety.

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Scope and Delimitation of the Study

The general intent of this study is to measure the viability of African

nightcrawlers (Eudrilus eugeniae) as a protein source to broiler chickens in terms of its

nitrogen free extract (NFE) and metabolizable energy (ME), and to compare its physical

properties; ash content, dry matter, and moisture content, chemical composition; crude

fat, crude fiber, and crude protein, and its NFE and ME to commercialized isolate soy

protein.

This study was conducted in Barangay Lalapac. Victoria, Tarlac. The researchers

raised, harvested, and processed the African nightcrawlers (Eudrilus eugeniae) in

Barangay Lalapac. Victoria, Tarlac. Safety and hygienic precautions were practiced in the

handling of the African nightcrawlers (Eudrilus eugeniae).

The testing for the ash content, dry matter, moisture content, crude fat, crude

fiber, crude protein, nitrogen free extract (NFE), and metabolizable energy (ME) was

carried out by the Region 3 Feed Chemical Analysis Laboratory in Barangay Sto. Niño,

San Fernando City, Pampanga.

The total timespan of this research is seven months.

Definition of Terms

African Nightcrawler. Also referred to as Eudrilus eugeniae. It is an earthworm species

commonly used in vermicomposting which has a distinctive mix of a gray and purple

color and grows to over twice the size of red worms, often reaching over 8 inches.

Ash Content. It indicates the amount of inorganic noncombustible material.

Broiler Chicken. Any chicken that is bred and raised specifically for meat production.

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Crude Fat. A traditional measure of fat in food products.

Crude Fiber. The insoluble residue of an acid hydrolysis followed by an alkaline one.

Crude Protein. A chemical analysis of the food whereby the amount of nitrogen present

is used to estimate the amount of protein in the food.

Dry Matter. A measurement of the mass of something when completely dried.

Earthworm. A terrestrial invertebrate that belongs to the phylum Annelida that breaks

down dead and decaying organic matter into rich humus soil.

Gross Energy (GE). It is the total amount of energy in an animal feed.

Hot Air Drying. A technique for drying foods in which heat is transmitted from hot air

to the product and evaporated water is carried to their air via convection.

Metabolizable Energy (ME). It is the net energy remaining after fecal and urinary

energy loss.

Moisture Content. It refers to the weight of the water contained in a certain object or

material.

Nitrogen Free Extract. It is designed to provide an estimate of water-soluble

polysaccharides (sugars, starch) and is calculated by the difference between the original

sample weight and the sum of weights of moisture (water), ether extract, crude protein,

crude fiber, and ash.

Poultry. Birds raised commercially or domestically for meat, eggs, and feathers.

Protein. Any of a class of nitrogenous organic compounds that consist of large molecules

composed of one or more long chains of amino acids and are an essential part of all living

organisms, especially as structural components of body tissues such as muscle, hair,

collagen, etc., and as enzymes and antibodies.

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Protein Source. The source of protein in animal feeds needed to nurture an animal’s

nutrition.

Push-Pull Method. A method of harvesting thru migration that consists of pushing the

earthworms out of the vermicompost of coconut leaves with the aid of repellant and

pulling them with attractant.

Vermicomposting. A process by which worms are used to convert organic materials

(usually wastes) into a humus-like material known as vermicompost.

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 CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDY

Related Literature

I. Foreign

Earthworms have been used for a long time. They have been eaten by early

humans and used as bait for fishing and hunting. Native Americans, Aborigines in

Australia and Maori in New-Zealand ate earthworms. Native Americans dried/smoked

and stored earthworms for winter use after they had been put in water or fed on special

feed in order to offload gut dirt and become tastier (Deane, 2020). Earthworms were

known for their ethnomedicinal properties in China and cited in the Divine Farmer’s

Materia Medica Classic around 200 B.C – 200 A.D (Yu Shen, 2010). In early

populations, earthworms were also already used as baits for fishing and hunting. By the

18th century, earthworms were referred to as a feedstock for domestic animals and Charles

Darwin was the first to document their importance for the breakdown of organic matter.

The industrial production of earthworms as fishing baits started in the 20th century in the

United States, notably in California. The use of earthworms as vermicomposters began in

the late 1930s, but this activity did not develop as these businesses were pyramid

schemes. A renewed interest arose in the 1970s in Germany, the United States and in the

United Kingdom. Today, the main use of earthworms is to produce vermicompost and to

provide protein for farm animals and fish. One of the advantages of vermicomposting is

that the process does not produce off-odors, unlike other composting processes.

Earthworms are able to decompose several classes of wastes coming from human, animal

or industrial activity (Edwards, 2011). Earthworms are edible and can be boiled, fried,

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roasted, sauteed, put in meatloaf and used in culinary specialties all over the world

(Deane, 2020).

Moreover, there are thousands of species of earthworms which can be

characterized into 3 major groups: anecic (deep-burrowing earthworms), endogeic

(upper-soil earthworms), and epigeic (surface-litter compost worms). The epigeic

compost worms are suitable and potential for scaling-up production of vermiculture as a

source of protein for animal feed from organic wastes because they have a high

reproduction, growth, and adaptation rates. At least there are 5 aspects that should be

taken into consideration in using compost worms as a source of animal feed from organic

wastes: Chemical composition, quality of protein value, potential hazards, production

economics, and legal constraints.

The chemical composition of compost worm is suitable for animal feed as the

composition of worm tissues consist of 80 – 90% of water and the dry matter basis which

are distributed into 60 – 70% of protein, 6 – 11% of fat, 5 – 21% of carbohydrate, and 2 –

3% of minerals with gross energy of 16 – 24 kilo Joule per gram.

The mean amounts of essential amino acids recorded from the composting worm

tissue are very adequate for good animal feed if compared with the recommendation of

the Food and Agriculture Organization (FAO) of the United Nations, particularly in terms

of lysine, methionine, cysteine, phenylalanine and tyrosine. In addition to containing

adequate protein content, earthworm tissues contain a preponderance of long-chain fatty

acids, many of which the non-ruminant animals cannot synthesize. They have an

excellent range of vitamins; are rich in vitamin B3 and provide a good source of vitamin

B12.

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 However, the potential hazards of using compost worms as animal feed have been

taken into consideration in case that they may carry diseases or contain toxic residues.

There are many studies have shown that the population of plant pathogens (such as

Fusarium, Pythium, Phytophthora) and animal or human pathogens (such as Salmonella,

E. coli, enteric viruses) can be significantly reduced by the presence of compost worms.

It is well known that compost worms produce anti-pathogen substances (antibacterial

peptides) called fetidin in their gut and pass through to the worm castings or

vermicompost. On the other hand, the beneficial soil microbes produced during

vermicomposting will suppress the pathogens. The toxic residues in the compost worms

can be limited using organic waste for example food waste, farm waste or green waste

with their sources are well-monitored and pre-composed in proper ways.

Furthermore, the economic return from compost worm production for animal feed

depends upon the rate and cost of production, and the price available for the product of

worm protein. There have been several studies of the benefits of the production of worm

protein. The general conclusions are that worm protein production has the best prospect

of good profits, if it is done by larger farmers with considerable amounts of available

unused organic wastes. The farmers may use the vermiculture – vermicomposting

technologies in separating worms from vermicompost to reduce the high cost of labor

involved and selling or using the by-product high quality vermicompost for their own

organic (natural) crops; to replace the application of the chemical fertilizers. Furthermore,

the purification of earthworms as a source of drugs i.e. as antioxidative, antiulcer,

anti-inflammatory, anticarcinogenic, anticoagulative have scientifically been studied by

15
the pharmaceuticals research for many years, and it will increase the value of the worm

protein.

Lastly, compost worms do not eat meat or dairy products. Concerning the legal

constraints of using compost worm protein as animal feed, if prepared by standard meat

e.g. the cooking and drying that converts abattoir waste to meat meal for animal feed or

by fish meal process, should meet little opposition, but again continual safety monitoring

will be necessary.

Related Studies

I. Foreign

Earthworm meal is made up of processed worms raised for vermicomposting,

which is a form of composting that involves the worms converting organic soil nutrients

for agriculture and horticulture from fruit and vegetable wastes, animal manure,

methanizer residues, or sewage into wormcast (castings). Vermicomposting produces

protein-rich earthworms as a byproduct. They can be fed to farm animals, particularly

poultry, pigs, and aquaculture. Earthworm breeding could help to reduce waste and

feed/food competition. From an economic and energy standpoint, earthworms may be

easier to raise than insects because many earthworm species are acclimated to a wider

temperature range than insects (Tedesco et al., 2019).

Earthworms come in over 4000 different species. Vermicomposting has been

limited to a few species because they have desirable traits such as tolerance to a wide

variety of environmental and management conditions, short life cycles, high reproduction

rates, and good composting rates. The only widely used species for vermicomposting are

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Eisenia andrei, Eisenia fetida (sometimes called Eisenia foetida), Dendrobaena veneta,

Perionyx excavatus, and Eudrilus eugeniae. Eisenia andrei and Eisenia fetida (red striped

worms) are two of the five species that are often mistaken. They can be found together in

the wild, however it is recommended that they be separated in vermicomposting systems

because mixing them diminishes their viability (Dominguez et al., 2011).

Furthermore, research on the usage of earthworms as an alternative protein source

dates back only 80 years. Earthworms can rapidly break down organic wastes (such as

cattle and pig manure) under controlled conditions to produce valuable horticultural

composts and high-grade protein suitable for animal feed (Edwards, 1985), and they are a

valuable potential source of animal feed able, on the one hand, to develop an efficient

management of agricultural waste and by-products and, on the other hand, to reintroduce

organic matter whose traditional management has been neglected in the food production

process (Hardy, 1996).

Son (2009) also found that supplementing laying hens' diets with EWM at levels

ranging from 0.2 to 0.6 percent increases laying performance and egg quality, particularly

the ratio of egg yolk n302 6/n-3 fatty acids. Furthermore, supplementing with 0.2 to 0.6

percent EWM increased broiler performance by increasing the digestibility of crude

protein in the diet.

On the other hand, at an international conference on the role of forests in food

safety and nutrition, the UN Food and Agriculture Organization presented a report titled

“Edible Insects: Future Prospects for Food and Feed Security” (Rome, 13-15 May, 2013).

According to the paper, eating some invertebrate creatures, such as earthworms, can help

battle global hunger and enhance global health standards by filling protein gaps and

17
lowering pollution. It was centered on the concept of entomological farming as a source

of animal feed and human sustenance.

Conceptual Framework

Figure 1. The Conceptual Framework

Figure 1 presents the conceptual framework of this study. It shows the input;

African nightcrawlers (Eudrilus eugeniae), the procedures to be taken: raising,

harvesting, and the processing of the African nightcrawlers (Eudrilus eugeniae) as a

protein source for broiler chickens, testing of the physical properties and chemical

18
composition of the African nightcrawlers (Eudrilus eugeniae) as a protein source,

calculation of the NFE and ME, and the statistical treatment, in order to achieve the

output; the viability of African nightcrawlers (Eudrilus eugeniae) as a protein source for

broiler chicken.

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 CHAPTER III

METHODOLOGY

Research Design

The research design used for this study is the two-group experimental design

wherein comparisons were done between the experimental group and the control group.

The African nightcrawlers (Eudrilus eugeniae) as a protein source served as the

experimental group while the commercialized isolate soy protein served as the control

group.

Procedure

This study employs the procedures done below in the viability of African

nightcrawlers (Eudrilus eugeniae) as a protein source for broiler chicken.

A. Raising of the African Nightcrawlers

Earthworm African nightcrawlers (Eudrilus eugeniae) were used to produce the

protein powder. African nightcrawlers have huge appetites, they will eat up to 150% of

their own body weight each day if given the proper environment and proper amount of

food. Among other composting worms, African nightcrawlers eat the most food per day.

They are also a larger breed of earthworm that gain weight easily, they can grow up to

8–10 inches long. African nightcrawlers produce approximately 2–3 cocoons per week

20
with 2–3 babies per cocoon averaging out to about 6–7 worms per week with their

breeding age as soon as 5 weeks. For these, they are the most promising earthworm to

use.

The earthworms were raised in a compost pit (2m x 3m x 1m depth) drained with

bamboo poles to prevent earthworms from being waterlogged. Rearing of earthworm

species requires a temperature ranging between 15 and 25 °C, soil moisture content

ranging between 60 and 85%, pH of 6.8 to 7.2 and can survive in a 163 relatively low O2

and high CO2 condition or in dissolved O2 within water (Sherman, 2003). The bed layer

was filled with organic residues (animal manure, peat moss, hay, papers, leaves, straw,

crop residues, etc.) beforehand and will be left for a week to precompost. The conversion

of animal wastes into earthworm tissue is very efficient: a ton of suitable animal wastes

produces up to 100 kg of worms, equivalent on a dry-mass basis to a conversion 166

efficiency in the order of 10% (Edwards, 1985). Hennuy and co-authors (1986) also

found similar results showing that 450 kg of vermicompost and 40 kg of earthworms (6.5

kg of 168 worm meal with 70% protein content for animal feed) were produced from 1

ton of cattle 169 manure.

The earthworms were then added after a week to start burrowing. Initial

earthworm density should be in 2.5–5 kg/m2 range to ensure an optimal reproduction rate.

The earthworm population under favorable conditions may double within 30–60 days

(Fosgate et al., 1972). Twenty-one kilograms of earthworms were added with three

kilograms of earthworm per meter squared of the pit. The pit was covered to prevent

sunshine and to keep the substrate with adequate moisture. Feeds were also distributed

regularly and in amounts small enough to prevent fermentation and a rise in temperature

21
that may affect the worms’ reproduction. After 60 days, the pit was partially (66–77%)

emptied and the worms were harvested. The vermicompost yield could be used as

fertilizer, to be sold or to be used.

B. Harvesting of the African Nightcrawlers

Gathering the earthworms can be done using different methods. However, the

push-pull migration method developed in India was utilized. This method consists of

pushing the earthworms out of the vermicompost of coconut leaves with the help of a

repellent (mustard) and to subsequently pull them with an attractant (cow dung). The

method was found to be effective and able to save manpower.

C. Processing the African Nightcrawlers for Animal Feeding

The earthworms were washed thoroughly and left in water for a duration of 30

minutes, so that they evacuated the residual wastes from their guts. They were later

blanched in boiling water and drenched in ice-cold water. Then, the earthworms were

dried through a dehydrator at 50°C for four hours. Finally, after the earthworms have

dried, they are grinded into powder using an electric grain grinder.

Data Gathering

To calculate the difference between African nightcrawlers (Eudrilus eugeniae)

and commercialized isolate soy protein as a protein source for broiler chicken, 200 grams

of each sample were sent to the Region 3 Feed Chemical Analysis Laboratory. A method

called complete proximate analysis was utilized to test for the dry matter, ash content,

moisture content, crude fat, crude fiber, and crude protein of the two groups.

22
Furthermore, the viability of the experimental group in comparison to the control group

as a protein source for broiler chicken in terms of nitrogen free extract (NFE) and

metabolizable energy (ME) were also tested by the Region 3 Feed Chemical Analysis

Laboratory. In testing for the NFE and ME, a complete proximate analysis is prerequisite.

Ash Content

To test for the ash content, a furnace-ignition method was utilized. A crucible was

weighed before and after 5g of the experimental group and control group sample were

added. After accurate weights are done, the crucible was then placed in the muffle

furnace for about 650°C until the particles of the samples turned into light gray ash.

Further after the observation of gray ash in the crucible, it was cooled in the desiccator at

room temperature and later on were weighed using analytical balance. Then, the mean

values were collected after weighing the samples three times each.

Moreover, the percentage presence of ash content was calculated using the

formula wherein the weight of the crucible plus ash (WOCPA) is subtracted by the

weight of the crucible (WOC) divided by the weight of the sample and multiplied by a

hundred.

𝑊𝑂𝐶𝑃𝐴 − 𝑊𝑂𝐶
% 𝑎𝑠ℎ = 𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡
× 100

Moisture Content

23
 To test the moisture content, the oven-drying method was used. Three grams of

each sample were kept in a hot air oven for an hour at 130°C. After drying, the samples

were placed in a desiccator and weighed using an analytical balance. This process was

done three times each sample and the mean values were collected. Finally, the moisture

content were calculated using the formula wherein the weight of the moisture (𝑊𝑚) is

divided by the weight of the bone dry material (𝑊𝑑) and multiplied by a hundred.

𝑊𝑚
𝑀 = 𝑊𝑑
× 100

Dry Matter

To test for the dry matter (DM), the formula of subtracting the moisture

percentage from 100 was used.

𝐷𝑀 = 100 − 𝑚𝑜𝑖𝑠𝑡𝑢𝑟𝑒 %

Crude Fat

To test for the crude fat content of the samples, a method called ANKOM™ filter

bag technology was utilized and calculations were made using the formula wherein a

hundred is multiplied to the difference of the weight of pre-dried sample with the filter

bag (𝑊2) subtracted by the weight of the dried sample and filter bag after extraction (𝑊3)

and divided by the original weight of the sample (𝑊1).

24
 100 × (𝑊2 − 𝑊3)
% 𝐶𝑟𝑢𝑑𝑒 𝐹𝑎𝑡 = 𝑊1

Crude Fiber

In order to test for the crude fiber content, the method ANKOM™ filter bag

technology was also used and calculations were made using the formula where the weight

of the organic matter (𝑊3) is subtracted by the product of the bag tare weight (𝑊1) and

the ash corrected blank bag factor (𝐶1) and divided by the weight of the sample (𝑊2) and

multiplied by a hundred.

[𝑊3 − (𝑊1 × 𝐶1)]

% 𝐶𝑟𝑢𝑑𝑒 𝐹𝑖𝑏𝑒𝑟 = 𝑊2
× 100

Crude Protein

To test the crude protein, Kjeldahl method will be utilized and calculated using

the formula wherein 6.25 is multiplied by the nitrogen content, in grams per kilogram, of

the test sample (𝑊𝑛).

% 𝐶𝑟𝑢𝑑𝑒 𝑃𝑟𝑜𝑡𝑒𝑖𝑛 = 6. 25 × 𝑊𝑛

Nitrogen Free Extract (NFE)

25
 To calculate the nitrogen free extract of the two samples, the formula wherein a

hundred was subtracted by the sum of crude fat, crude fiber, crude protein, ash content,

and moisture content was used.

𝑁𝐹𝐸 = 100 − (𝑐𝑟𝑢𝑑𝑒 𝑓𝑎𝑡 + 𝑐𝑟𝑢𝑑𝑒 𝑓𝑖𝑏𝑒𝑟 + 𝑐𝑟𝑢𝑑𝑒 𝑝𝑟𝑜𝑡𝑒𝑖𝑛

+ 𝑎𝑠ℎ 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 + 𝑚𝑜𝑖𝑠𝑡𝑢𝑟𝑒 𝑐𝑜𝑛𝑡𝑒𝑛𝑡)

Metabolizable Energy (ME)

In order to test and evaluate the metabolizable energy, the gross energy (GE) of

the samples were first calculated. To calculate for the GE, the formula is as follows:

𝐺𝐸 = (5. 7 × 𝑐𝑟𝑢𝑑𝑒 𝑝𝑟𝑜𝑡𝑒𝑖𝑛) + (9. 4 × 𝑐𝑟𝑢𝑑𝑒 𝑓𝑎𝑡) +

[4. 1 × (𝑁𝐹𝐸 + 𝑐𝑟𝑢𝑑𝑒 𝑓𝑖𝑏𝑒𝑟)]

Then, the GE of each sample was converted to ME. It was done by multiplying

the GE by 0.82. A study by Cerniglia (1981) had shown that the fecal and urinary energy

loss in broiler chicken is 18%, therefore, the metabolizable energy in broiler chicken is

82% of the total gross energy.

𝑀𝐸 = 𝐺𝐸 × 0. 82

26
Statistical Treatment

The statistical treatment used in this study is the independent t-test, since it

compares the mean of two samples, which are the African nightcrawlers (Eudrilus

eugeniae) as a protein source for broiler chicken as the experimental group and the

commercialized isolate soy protein as the control group.

Risk and Safety

As the purpose of this study is to measure the viability of African nightcrawlers

(Eudrilus eugeniae) as a protein source for broiler chicken in comparison to a

commercialized isolate soy protein, the handling of the protein powders was done in

consideration of safety and hygiene.

In the process of processing the African nightcrawlers (Eudrilus eugeniae) as a

protein source, hygienic practices were observed. The researchers wore personal

protective equipment (PPE) such as gloves and hairnets. Furthermore, the equipment

used to process the earthworms were washed thoroughly before and after the process. The

final product was sealed in an air-tight ziplock bag and stored properly.

During the process of testing, professional laboratory technologists performed

safety and hygienic practices in accordance with the facility’s risk and safety protocols.

The laboratory materials and equipment were disinfected before and after usage in order

to avoid cross-contamination with other test samples. Moreover, the wearing of PPE was

practiced.

27
 CHAPTER IV

RESULTS AND DISCUSSION

This chapter presents the results from the study, and evaluated the viability of

African nightcrawlers (Eudrilus eugeniae) as a protein source for broiler chicken in

comparison to commercialized isolate soy protein in terms of their physical properties,

chemical composition, nitrogen free extract (NFE) and metabolizable energy (ME).

Table 1. Physical Properties of African Nightcrawlers (Eudrilus eugeniae)

Physical Properties Value (% per 100 grams)

Ash Content 4.54

Moisture Content 6.42

Dry Matter 93.58

Table 1 presents the values of the physical properties of African nightcrawlers

(Eudrilus eugeniae) in percentage per 100 grams using three parameters; ash content,

moisture content, and dry matter. The ash content of African nightcrawlers (Eudrilus

eugeniae) is at 4.54 percent, the moisture content is at 6.42 percent, and the dry matter is

at 93.58 percent.

Table 2. Chemical Composition of African Nightcrawlers (Eudrilus eugeniae)

Chemical Composition Value (% per 100 grams)

Crude Fat 7.44

Crude Fiber 6.43

Crude Protein 62.60

28
 The table above shows the value of the chemical composition of African

nightcrawlers (Eudrilus eugeniae) in percentage per 100 grams using three parameters;

its crude fat, crude fiber, and crude protein composition. Based on the table, African

nightcrawlers (Eudrilus eugeniae) have 7.44 percent of crude fat, 6.43 percent of crude

fiber, and 62.60 percent of crude protein.

Table 3. Physical Properties of Commercialized Isolate Soy Protein

Physical Properties Value (% per 100 grams)

Ash Content 6.32

Moisture Content 9.78

Dry Matter 90.22

The table presents the values of the physical properties of the commercialized

isolate soy protein in percentage per 100 grams in terms of its ash content, moisture

content, and dry matter. Table 3 shows that the commercialized isolate soy protein has a

total of 6.32 percent of ash content, 9.78 percent of moisture content, and 90.22 percent

of dry matter.

Table 4. Chemical Composition of Commercialized Isolate Soy Protein

Chemical Composition Value (% per 100 grams)

Crude Fat 19.23

Crude Fiber 4.43

Crude Protein 40.62

29
 Table 4 shows the chemical composition values of the commercialized isolate soy

protein in percentage per 100 grams in terms of its crude fat, crude fiber, and crude

protein content. The table presents that the commercialized isolate soy protein is

primarily composed of 19.23 percent of crude fat, 4.43 percent of crude fiber, and 40.62

percent of crude protein.

Table 5. Nitrogen Free Extract (NFE) of African Nightcrawlers (Eudrilus eugeniae) and

Commercialized Isolate Soy Protein

Total Nitrogen Free

Value (% per
Sample Parameters Extract (NFE) (%
100 grams)
per 100 grams)

Ash Content 4.54

African
Moisture Content 6.42
nightcrawlers
Crude Fat 7.44 12.57
(Eudrilus eugeniae) Crude Fiber 6.43

Crude Protein 62.60

Ash Content 6.32

Commercialized Moisture Content 9.78

Crude Fat 19.23 19.60

Isolate Soy Protein
Crude Fiber 4.43

Crude Protein 40.62

The table above shows the nitrogen free extract (NFE) content of African

nightcrawlers (Eudrilus eugeniae) and the commercialized isolate soy protein, as well as

their ash content, moisture content, crude fat, crude fiber, and crude protein in percentage

per 100 grams.

30
 The total NFE content of African nightcrawlers (Eudrilus eugeniae) is 12.57

percent per 100 g with its ash content at 4.54 percent, moisture content at 6.42 percent,

crude fat at 7.44 percent, crude fiber at 6.43 percent, and crude protein at 62.60 percent.

On the other hand, the commercialized isolate soy protein has a total NFE content

of 19.60 percent, ash content of 6.32 percent, moisture content of 9.78, crude fat content

of 19.23 percent, crude fiber of 4.43 percent, and crude protein of 40.62 percent.

Table 6. Metabolizable Energy (ME) of African Nightcrawlers (Eudrilus eugeniae) and

Commercialized Isolate Soy Protein

Total
Metabolizable
Value (per
Sample Parameters Energy
100 grams)
(kilocalories per
100 grams)

Gross Energy 504.66 kcal

African Nitrogen Free
12.57%
Extract
nightcrawlers 413.82
Crude Fat 7.44%
(Eudrilus eugeniae)
Crude Fiber 6.43%

Crude Protein 62.60%

Gross Energy 510.82 kcal

Nitrogen Free
Commercialized 19.60%
Extract
418.87
Isolate Soy Protein Crude Fat 19.23%

Crude Fiber 4.43%

Crude Protein 40.62%

31
 Table 6 presents the metabolizable energy (ME) content and the five parameters

that compose the total ME; gross energy (GE), nitrogen free extract (NFE), crude fat,

crude fiber, and crude protein per 100 grams.

The table revealed the total ME content of African nightcrawlers (Eudrilus

eugeniae) is 413.42 kcal/100g and is composed of 504.66 kcal/100g GE, 12.57 percent

NFE, 7.44 percent crude fat, 6.43 percent crude fiber, and 62.60 percent crude protein.

Furthermore, table 6 also presented that the commercialized isolate soy protein

has a total ME content of 418.87 kcal/100g, GE of 510.82 kcal/100g, NFE of 19.60

percent, crude fat of 19.23 percent, crude fiber of 4.43 percent, and crude protein of

40.62 percent.

Statistical Analysis Report of Nitrogen Free Extract (NFE)

Table 7. Independent T-Test of African Nightcrawlers (Eudrilus eugeniae) and

Commercialized Isolate Soy Protein in Terms of Nitrogen Free Extract (NFE), Ash

Content, Moisture Content, Crude Fat, Crude Fiber, and Crude Protein

Parameters Significant Difference

Nitrogen Free Extract (NFE) 0.137

Ash Content 0.103

Moisture Content 0.130

Crude Fat 0.265

Crude Fiber 0.116

Crude Protein 0.134

32
 The table above shows the results of the independent t-test conducted to compare

the two groups; the African nightcrawlers (Eudrilus eugeniae) and the commercialized

isolate soy protein in terms of their nitrogen free extract (NFE) content, ash content,

moisture content, crude fat, crude fiber, and crude protein.

The data computed presented the significance value of the two groups in terms of

NFE at 0.137, ash content at 0.103, moisture content at 0.130, crude fat at 0.265, crude

fiber at 0.116, and crude protein at 0.134. Since the p-value calculated for all the

parameters are higher than the alpha level of 0.05, it can be inferred that the null

hypothesis must be accepted. Therefore, there were no significant differences between the

NFE content and its composition of the two groups.

Statistical Analysis Report of Metabolizable Energy (ME)

Table 8. Independent T-Test of African Nightcrawlers (Eudrilus eugeniae) and

Commercialized Isolate Soy Protein in Terms of Metabolizable Energy (ME), Gross

Energy (GE), Nitrogen Free Extract (NFE), Crude Fat, Crude Fiber, Crude Protein

Parameters Significant Difference

Metabolizable Energy (ME) 0.004

Gross Energy (GE) 0.004

Nitrogen Free Extract (NFE) 0.137

Moisture Content 0.130

Crude Fat 0.265

Crude Fiber 0.116

Crude Protein 0.134

33
 The table above presents the significant difference between the metabolizable

energy (ME) and its constituents; gross energy (GE), nitrogen free extract (NFE),

moisture content, crude fat, crude fiber, and crude protein of the two groups when

statistically analyzed using independent t-test.

The data computed for the significance level of ME and GE is 0.004. On the other

hand, NFE has a significance level of 0.137, ash content has 0.103, moisture content has

0.130, crude fat has 0.265, crude fiber has 0.116, and crude protein has 0.134. Since NFE,

ash content, moisture content, crude fat, crude fiber, and crude protein has a p-level

higher than the alpha level of 0.05, it can be concluded that there was no significant

difference between the two groups in these parameters. In opposition, the p-level of ME

and GE is lower than the alpha level of 0.05, therefore, it can be inferred that there were

significant differences between the two groups in these parameters. Therefore, the null

hypothesis must be rejected.

Discussion

The physical properties of African nightcrawlers (Eudrilus eugeniae) were found

to be appropriate for broiler chicken feeding. The ash content of African nightcrawlers

(Eudrilus eugeniae) is suitable for feeding since it falls within the standard ash content in

poultry feed which is from 5 – 8 percent. Keeping the ash content at the standard range

ensures the digestibility of the feeds. Similarly, the moisture content is acceptable under

the standard moisture content of livestock feeding of not more than 11.5 percent.

Ensuring that the moisture content falls under the standard requirements are important for

the feed’s shelf-life and resistance to bacterial inhibition. The appropriateness of the

34
physical properties suggests that African nightcrawlers (Eudrilus eugeniae) are viable to

be used as a protein source for broiler chickens.

The chemical composition of African nightcrawlers (Eudrilus eugeniae) were not

found to be completely suitable for broiler feeding. The crude fat content is suitable

under the standard 5 – 7.5 percent requirement for broiler chickens. It is crucial to keep

the crude fat under the standard requirements since it affects the broiler’s performance

such as the egg production rate and the quality of the eggs. Furthermore, the crude fiber

content was found to be appropriate as it is kept below 7 percent which is the standard

requirement. Keeping the crude fiber below 7 percent encourages healthy digestion and

nutrient absorption in broilers. On the other hand, the crude protein content exceeds the

standard 15 to 19 percent requirement. However, the crude protein can be controlled with

a balanced diet.

The nitrogen free extract (NFE) content of African nightcrawlers (Eudrilus

eugeniae) cannot be determined whether appropriate or not since there were no standard

requirements established. However, statistical analysis showed that there were no

significant differences between the NFE content of African nightcrawlers (Eudrilus

eugeniae) and commercialized isolate soy protein. This indicates that the African

nightcrawlers (Eudrilus eugeniae) could be compared to commercialized isolate soy

protein in terms of NFE.

The metabolizable energy (ME) content of African nightcrawlers (Eudrilus

eugeniae) exceeds the standard requirement of 3,000 kcal/kg to 3,200 kcal/kg. This

suggests that the ME content of African nightcrawlers (Eudrilus eugeniae) may not be

suitable for feeding. Statistical analysis also showed that the ME of African nightcrawlers

35
(Eudrilus eugeniae) is significantly lower than that of commercialized isolate soy protein.

This indicates that the African nightcrawlers (Eudrilus eugeniae) could not be compared

to commercialized isolate soy protein in terms of ME.

36
 CHAPTER V

CONCLUSION AND RECOMMENDATIONS

Raising poultry requires adequate nutrition and feeding with protein source being

one of the most important aspects as a feed ingredient. Having adequate protein sources

for broiler chickens is essential to their overall performance, however, most of the ones

available for commercial use are not sustainable for the environment. Given these, it

brought the researchers to explore various alternative sources such as vermicomposting

worms which possess the most promising potential, concluding with the utilization of

African nightcrawlers (Eudrilus eugeniae). Ergo, this study aimed to investigate the

viability of African nightcrawlers (Eudrilus eugeniae) as a protein source for broiler

chicken.

Using independent t-test as this study’s statistical treatment, this study compared

the viability of African nightcrawlers (Eudrilus eugeniae) with commercialized isolate

soy protein in terms of nitrogen free extract (NFE) together with its components; ash

content, moisture content, crude fat, crude fiber, and crude protein, and metabolizable

energy (ME) with its components; gross energy (GE), nitrogen free extract (NFE),

moisture content, crude fat, crude fiber, and crude protein. The results revealed that while

there were no significant differences between the two groups in terms of NFE as well as

its components, the ME and GE contents differ significantly.

The results lead to the acceptance of the null hypothesis that there are no

significant differences between African nightcrawlers (Eudrilus eugeniae) and

commercialized isolate soy protein in terms of nitrogen free extract (NFE). At the same

37
time, it is also concluded that there are significant differences between the two groups in

terms of metabolizable energy (ME). This study not only shed light to the promising

potential of vermicomposting worms, specifically African nightcrawlers (Eudrilus

eugeniae), as a protein source for poultry feeding but also in unlocking new knowledge

that offers insights and builds foundation for future research, practice, or policy.

Conclusion

1. The physical properties of the African nightcrawlers (Eudrilus eugeniae) are

adequate and acceptable for poultry feeding. This suggests that African

nightcrawlers (Eudrilus eugeniae) could be viable as a protein source for broiler

chicken.

2. The chemical composition of African nightcrawlers (Eudrilus eugeniae) is not

completely appropriate for broiler feeding since one of its components exceeds

the standard requirement for broiler feeding. However, it could be controlled by a

balanced diet. This suggests that African nightcrawlers (Eudrilus eugeniae) could

be viable as a protein source for broiler chicken with adequate diet.

3. The appropriateness of the nitrogen free extract (NFE) content of African

nightcrawlers (Eudrilus eugeniae) could not be determined. However, statistical

analysis showed that African nightcrawlers (Eudrilus eugeniae) and

commercialized isolate soy protein are similar in terms of NFE with significant

difference at 0.137.

38
 4. The metabolizable energy (ME) content of African nightcrawlers (Eudrilus

eugeniae) were found to exceed the nutritional requirements in broiler feeding.

However, statistical analysis concluded that African nightcrawlers (Eudrilus

eugeniae) and commercialized isolate soy protein differ in terms of ME with

significant difference at 0.004.

Recommendations

The researchers have come up with the following recommendations for future

investigation after compiling the data that was gathered and interpreting the results that

were achieved;

1. To increase the yield of protein powder from African nightcrawlers (Eudrilus

eugeniae), instead of using the dehydration method via a dehydrator, it is

recommended to explore other methods of drying such as freeze-drying via a

freeze-drier.

2. To analyze the complete chemical composition of the African nightcrawlers

(Eudrilus eugeniae) as a protein source, it is conferred to test for the mineral

contents of the product such as phosphorus, calcium, and salt.

3. In order to ensure the safety of utilizing African nightcrawlers (Eudrilus

eugeniae) as a protein source, it is suggested to test for the health hazards and

pathogen/toxin content of the product. Moreover, it is highly commended to test

for the levels of heavy metals such as arsenic (As), cadmium (Cd), chromium

(Cr), mercury (Hg), lead (Pb), copper (Cu), zinc (Zn), and nickel (Ni) as they are

39
 the most common pollutants in soil. Testing for the coliform count is also highly

recommended.

4. To evaluate the efficiency of the African nightcrawlers (Eudrilus eugeniae) as a

protein source, it is recommended to conduct an in vivo experiment wherein the

protein powder will be fed to broiler chickens with adequate feed formulation

based on their nutritional requirements. This could involve the investigation of the

growth rate, egg production rate, and the egg quality in relation to the African

nightcrawlers (Eudrilus eugeniae) as a protein source.

5. Finally, additional research is highly recommended in order to utilize the

maximum potential of African nightcrawlers (Eudrilus eugeniae) and/or other

earthworm species as a protein source for broiler chickens. This could involve

investigating the formulation of African nightcrawlers (Eudrilus eugeniae) and/or

other earthworm species as a protein source as a feed ingredient and/or as a

supplementary protein source for broiler chickens.

40
APPENDICES

41
 REFERENCES

Antonava, E., Titov, I., Pashkova I., Stom, D. (2021). Vermiculture as a source of animal

protein. E3S Web of Conferences 254, 08006 (2021).

https://doi.org/10.1051/e3sconf/202125408006

AOCS Approved Procedure Ba 6a-05. (n.d.).

http://www.ssco.com.tw/Ankom/PDF_file/Crude%20Fiber%20Method%20A200.

pdf

Bennett, G. L., Coppin, C. A., Smith, Z., McDaniel, Z. S., Tedeschi, L. O., Cole, N. A., &

Galyean, M. L. (2022). Predicting Metabolizable Energy from Digestible Energy

for Growing and Finishing Beef Cattle and Relationships to Prediction of

Methane. https://doi.org/10.1093/jas/skac013

Bogdanov, P. (1996). Commercial vermiculture: How to build a thriving business in

redworms. VermiCo Press, 1996, 83 p.

https://www.amazon.fr/dp/0965703908/ref=rdr_kindle_ext_tmb

Cerniglia, G. J. (1981). The Metabolizable Energy Determination of Fats and Oils in The

Metabolizable Energy Determination of Fats and Oils in Broiler Chickens.

Broiler Chickens.

https://digitalcommons.lsu.edu/cgi/viewcontent.cgi?article=4587&context=gradsc

hool_disstheses

Deane, G. (2020). Earthworms. Eattheweeds website.

http://www.eattheweeds.com/cooking-with-earthworms-2/

42
Edwards, C. A., Arancon, N. Q., Sherman, R. (2011). Vermiculture technology:

earthworms, organic wastes, and environmental management. CRC Press, Taylor

& Francis Group, Boca Raton, FL, USA

Fosgate, O. T.; Babb, M. R., 1972. Biodegradation of animal waste by Lumbricus

terrestris. J. Dairy Sci., 55 870-872

Ganzaroli, A., Bacenetti, J., & Parolini, M. (2020, September 10). Earthworm as an

alternative protein source in poultry and fish farming: current applications and

future perspectives. https://core.ac.uk/display/333583482

Gopal, M; Gupta, A. (2019). A simple push–pull strategy to harvest earthworms from

coconut leaf vermicompost produced in tanks. Current Science, 117 (10):

138-144. https://doi.org/10.18520/cs/v117/i1/138-144

Gunadi, B. (2015, December 1). Compost worms as a source of protein for animal feed.

burnabyredwigglers.

https://burnabyredwigglers.wordpress.com/2015/12/01/compost-worms-as-a-sour

ce-of-protein-for-animal-feed/

Heuzé V., Tran G., Sauvant D., Bastianelli D., Lebas F. (2020). Earthworm meal.

Feedipedia, a programme by INRAE, CIRAD, AFZ, and FAO.

https://www.feedipedia.org/node/665

Misra, R.V.; Roy, R.N.; Hiraoka, H. (2003). On-farm composting methods. Land Water

Disc. Paper, FAO Rome, Italy. http://www.fao.org/3/a-y5104e.pdf

Munroe, G. (2008). Guide du lombricompostage et de la lombriculture à la ferme. Centre

d’agriculture biologique du Canada.

https://www.agrireseau.net/documents/Document_96310.pdf

43
Nutrient Requirements of Poultry. (n.d.).

https://www.agropustaka.id/wp-content/uploads/2020/04/agropustaka.id_buku_N

utrient-Requirements-of-Poultry_Ninth-Revised-Edition-1994-NRC.pdf

Obwogo, S. (2020, June 29). Diary of a poultry farmer: Feeding regime for chicks

important.

https://nation.africa/kenya/business/seeds-of-gold/diary-of-a-poultry-farmer-feedi

ng-regime-for-chicks-important-424692

Stricklan, K., & Strickland, J. (2018). The Amazing African Nightcrawlers.

https://midwestworms.com/blogs/worm-blog/the-amazing-african-nightcrawlers

Yu Shen. (2010). Earthworms in traditional Chinese medicine. Zool. Middle East 51

(sup2): 171-173. https://doi.org/10.1080/09397140.2010.10638470

44