CHAPTER ONE

1.1 INTRODUCTION

The industrialization of poultry husbandry and the improvement of feed nutritional efficiency

have accelerated the introduction of feed additives which became widely used in animal feed for

many decades. The objective outlined by scientists, is to increase production (eggs & meat)

while maintaining animals in good health (Alloui et al., 2014). The use of antibiotics in poultry

feed as a growth promoter is beneficial in the improvement of production parameters and

diseases prevention. However this large utilization has led to the increasing resistance of

pathogens to antibiotics and the accumulation of antibiotic residues in animal products and in the

environment (Alloui et al., 2014). This situation requires the world to restrict using AGPs in

animal feed (Nisha, 2008). The removal of AGPs from animal feed may affect their productions

performance and foster the resurgence of pathogens causing illness and economic losses in

farms. In this context, herbs and plant extracts are searched to be incorporated in poultry feed as

growth promoters such as probiotics and prebiotics (Alloui, 2013).

Feed additives can either be nutritive or non-nutritive products added to the based diet, and are

minor components of the animal diet. Feed additives are products used in animal nutrition for the

purposes of improving the quality of feed and the quality of food from animal origin, improve

the animal’s performance and health, e.g. providing enhanced digestibility of the feed materials.

(Mandey and Sompie, 2021) Feed additives promote ingestion, absorption, assimilation of

nutrients, growth, and health by affecting the physiological processes, such as immune function

and stress resistance. Feed additives include immune stimulants, prebiotics, probiotics, acidifiers,

essential oils, or others. (Mandey and Sompie, 2021). Some of the commonly feed additives in

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animal diets include enzymes, pro- and prebiotics, antioxidants, antibiotic growth promoters, and

colouring agents.

Phytogenics are a group of natural growth promoters (NGPs) or non-antibiotic growth promoters

used as feed additives, derived from herbs, spices or other plants (e.g. garlic, oregano, thyme,

rosemary, coriander and cinnamon) as well as to their respective plant extracts in the form of

essential oils (Windisch et al., 2008). They are commonly regarded as favourable alternatives

feed additives to antibiotic growth promoters (AGPs) in poultry production (Windisch et al.,

2008). The supplementation of phytogenics in poultry diets is becoming more popular as it is

cheap and readily available, less toxic, rich in nutrients and useful in improving the health of

animals and consumers (Dhama et al., 2015). Phytogenics enhance amino acid availability and

increase performance in body weight of the birds, carcass traits, and health status and limit the

adverse effects of antioxidative stress and ammonia emissions in birds (King, 2017; Valenzuela

Grijalva et al., 2017; Oloruntola et al., 2018) as well as lower cholesterol content thereby

improving meat quality (Oloruntola et al., 2018; Oloruntola et al., 2019).

Turmeric (Curcuma longa) is one of the numerous phytogenic additives of importance in poultry

feed production (Basak, 2015). Turmeric is a rhizome of the herbaceous perennial plant of the

ginger family, Zingiberaceae. Though it is known to be native to the tropical South Asia, it is

also widely grown in Nigeria and other tropical and sub-tropical Africa since it requires

temperature between 20 and 30℃ and a considerable amount of annual rainfall for growth (Khan

et al., 2012). It is a domestic spice has the various applications in the medicinal biology

(Aggarwal and Harikuma, 2009). Turmeric produces a specific bioactive compound called

curcumin, a polyphenolic phytochemical with anti-microbial, anti-inflammatory, anti-cancerous

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and anti-oxidant properties (Al-Sultan, 2003; Aggarwal and Harikuma, 2009). The proximate

compositions of turmeric rhizome powder was revealed to have an appreciable amounts of crude

protein (10.07%), ash (2.76%), crude fibre (4.87%), ether extract (6.64%) and nitrogen free

extract (66.76%) (Imoru et al., 2018).

Syzygium aromaticum (clove) is an aromatic plant commonly used as a spice. It is an evergreen

plant available all year round with different harvest seasons in different countries (Yun, 2018).

Aside from vitamins A, K, B6, B1 and C present in S. aromaticum (Dorman and Deans, 2000). It

contains numerous biologically active compounds such as eugenol (72–90%), eugenol acetate, β

caryophyllene, flavonoids and triterpenoids (Bhowmik et al., 2012; Jimoh et al., 2017). It has

been well reported for its antimicrobial properties (Wang and Kim, 2011) and immune

stimulating properties which have positive effects on the growth performance and health of

poultry (Chowdhury et al., 2018; Kunnumakkara et al., 2018, Al- Mufarrej et al., 2019). Clove

and its essential oil are highly relevant in poultry to improve growth performance by enhancing

the intestinal microbiota population (Mohammadi et al., 2014). The proximate chemical

composition of clove contains 10.0 % moisture, 20.0% crude fibre, 5.2% ash, 12.1%fat, 51.5%

carbohydrate and 1.2% crude protein (Abdel Moneim et al., 2007).

Serum protein refers to the total amount of protein present in the liquid portion of blood (i.e., the

serum). It is an important indicator of the health and nutritional status of an organism, as well as

a key diagnostic tool for many diseases and disorders. In animals, including chickens, serum

protein levels can be affected by a variety of factors, including diet, age, breed, and health status.

Serum protein levels are typically measured using laboratory tests, and the results are reported as

a reference range.

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1.2 GENERAL OBJECTIVE OF THE STUDY

The study aims at evaluating laying performance, nutrient digestibility and serum proteins of

laying hens fed dietary turmeric and clove powder.

1.3 SPECIFIC OBJECTIVES OF THE STUDY

The specific objectives of the study are to:

1. Analyze proximate composition of experimental diets, clove and turmeric power

2. Assess laying performance of hens fed turmeric and clove powder

3. Evaluate nutrient digestibility of laying hen fed experimental diets

4. Determine serum proteins of laying hens fed experimental diets supplemented with

turmeric and clove powder

5. Evaluate main and interaction effects of turmeric and clove powder on laying

performance and blood profile of hens.

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 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 POULTRY PRODUCTION

Livestock production constitutes a very important component of the agricultural economy of

developing countries, a contribution that goes beyond direct food production to include

multipurpose uses, such as skins, fibre, fertilizer and fuel, as well as capital accumulation.

Furthermore, livestock are closely linked to the social and cultural lives of several million

resource-poor farmers for whom animal ownership ensures varying degrees of sustainable

farming and economic stability (Okoro, 2016). Nutrition is the most expensive factor in poultry

production taking approximately 70% of production budgets (Mmadubuike and Ekenyem, 2001).

Therefore, a reduced cost of production while improving the feed efficiency would be a feasible

option. Various feed additives are used in poultry to maximize net returns and carcass quality. In

the past, growth-promoting antibiotics were used as feed additives; however, these were

associated with residues in the meat and eggs by consumers, and have been banned or limited in

many countries (Diarra et al., 2011). As a result, natural alternatives to antibiotics, such as herbs

and medicinal plants, have attracted attention due to their wide range of potential beneficial

effects (Mahesh and Prabhakar 2018).

2.2 NATURAL FEED ADDITIVES

The awareness in feed additives flourished over the few decades. Feed additives are a cluster of

nutrients and non-nutrient composites which help in improving the efficiency of feed utilization

and consequently dropping the high cost of feed (Akhtar et al., 1984). Feed additives are also

5
minor components of the animal ration and are used for improving the quality/digestibility of

feed and the nutritive and aesthetic quality of food or improving animal performance and health

(Cherian Gita, 2022). These additives have established a great consideration as feed supplements

for numerous purposes in poultry production throughout the recent years (Al-Amin et al., 2006).

In the past, antibiotics were regularly used as feed additives (Ali et al., 2008). Though, currently

use of antibiotics is not only restricted but also their practice in livestock and poultry industry

have been prohibited in many countries due to modification of natural gut microbiota and drug

resistance in microorganisms and humans. Natural growth promoters such as prebiotics,

probiotics, symbiotic, enzymes, plant extracts, etc., can be used to feed the broilers without any

adverse effect on the performance of birds (Al-Mashhadani, 2015). Beneficial properties of

bioactive plant constituents in animal nutrition may comprise the stimulation of appetite and feed

intake, the enhancement of endogenous digestive enzyme secretion, stimulation of immune

response and antibacterial, antiviral and antioxidant action (Al-Sultan, 2003).

Phytoadditives or phytogenic feed additives in animal nutrition have attracted a lot of attention

for their potential role as alternatives to antibiotic growth promoters. Phytoadditives are feed

additives originated from plants or botanicals that are used in poultry nutrition. These substances

are derived from herbs, spices, and other plants and their extracts. They are natural, less toxic,

residue free and ideal feed additives for poultry when compared to synthetic antibiotics (Mandey

and Sompie, 2021). There efficacy of phytogenic applications in poultry nutrition depends on

several factors, such as composition and feed inclusion level of phytogenic preparations, bird

genetics, and overall diet composition (Mandey and Sompie, 2021). Phytoadditives have

antimicrobial, antifungal, antiviral, antitoxigenic, antiparasitic and insecticidal properties. The

benefits of using phytoadditives in poultry nutrition are increased feed intake, stimulation of

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digestion increased growth performance, reduced incidence of disease, improved reproductive

parameters, feed efficiency, profitability (Mandey and Sompie, 2021).

Commonly used to classify the vast variety of phytogenic compounds, mainly with respect to

origin and processing, such as herbs (flowering, non woody, and non-persistent plants), spices

(herbs with an intensive smell or taste commonly added to human food), essential oils (volatile

lipophilic compounds derived by cold expression or by steam or alcohol distillation), or

oleoresins (extracts derived by non-aqueous solvents) (Yitbarek, 2015). Within phytogenic feed

additives, the content of active substances in products may vary widely, depending on the plant

part used (e.g., seeds, leaf, root, or bark), harvesting season, and geographical origin. The

technique for processing (e.g., cold expression, steam distillation, extraction with non-aqueous

solvents, etc.) modifies the active substances and associated compounds within the final product

(Yitbarek, 2015).

2.3 ORIGIN AND DESCRIPTION OF TURMERIC

One of the many medicinal herbs promising agricultural products as natural feed additives in

poultry diets is Turmeric (Curcuma longa Linn.) from the Zingiberaceae family. It is a perennial

plant with a short stem and large oblong leaves, and it bears ovate, pyriform, or oblong rhizomes,

which are often branched and brownish-yellow in colour (Daneshyar et al., 2011). Though it is

known to be native to the tropical South Asia, it is also grown in other tropical and sub-tropical

Africa including Nigeria. It requires temperature between 20 and 30 ℃ and a considerable

amount of annual rainfall for growth (Khan et al., 2012). Curcumin is the active compound in

turmeric powder which is reported to have a lot of molecular targets in the cells (Zhou et al.,

2012) that could affect cell functions in the body.

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2.3.1 Taxonomy of Turmeric

Turmeric is taxonomically classified below;

Kingdom - Plantae

Class - Liliospida

Sub-class - Commelinids

Order - Zingiberales

Family - Zingiberaceae

Genus - Curcuma

Species – Curcuma longa

The wild turmeric is called C. aromatic and domestic species is called C. longa. (Shrishail et al.,

2013)

2.3.2 Phytocomponents of Turmeric

The medicinal properties of turmeric, the supply of curcumin, were regarded from many years;

but, the capability to decide the exact mechanism(s) of action and to decide the bioactive

compounds have only these days been investigated as follows: (Gupta et al., 2013). Curcumin is

also referred as diferuloyl methane, is the principle herbal polyphenol observed inside the

rhizome of Curcuma longa (turmeric) and in others Curcuma spp (Aggarwal et al., 2003).

Curcuma longa has been historically used in Asian countries as a medical herb because of its

antioxidant, antimutagenic, antimicrobial (Mahady et al., 2002; Reddy et al., 2005), and

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anticancer properties (Vera-Ramirez et al., 2013; Wright et al., 2013). Turmeric contains a high

level of beneficial phenolic compounds (curcuminoids) and terpenoids (sesquiterpenes). The

main curcuminoids of the rhizome are curcumin, demethoxycurcumin, and

bisdmethoxycurcumin. Whereas ar turmerone, α-turmerone, and β-turmerone are the major

ketonic sesquiterpenes of turmeric essential oil. These compounds have been reported to have

antibacterial, antioxidant, anti-inflammatory, antihepatotoxic, anticarcinogenic, and

hypocholesterolemic activities (Nishiyama et al., 2005; Daneshyar et al., 2011; Li et al., 2011;

Abou-Elkhair, 2014; Alagawany et al., 2016).

The energetic parts of turmeric are the flavonoid Curcuminoids that aggregates curcumin

(diferuloyl methane), mono dexmethoxy curcumin and bisdesmethoxy curcumin. Curcumin

makes up approximately 90% of the curcuminoid content material in turmeric. Other elements

encompass sugars, proteins, and resins. The best researched energetic constituent is curcumin,

which comprises 35.4% of uncooked turmeric (Heath et al., 2004). Turmeric incorporates 69.4%

carbohydrates, 6.3% protein, 5.1% fats, 3.5% minerals, and 13.1% Moisture. The crude oil

(5.8%) acquired by way of steam distillation possesses quiterpenes (53%), zingiberene (25%),

aphellandrene (1%), sabinene (0.6%), cineol (1%), and borneol (0.5%). Curcumin (3–4%) is

responsible for the yellow color, and incorporates curcumin I (94%), curcumin II (6%) and

curcumin III (3%) (Ammon et al., 1991). Curcumin is hydrophobic phenol and is responsible for

the turmeric’s orange-yellow colour (Tanzeela et al., 2015; Choudhury, 2019). The primary issue

of curcumin is its poor absorption in the small intestine. Nevertheless, novel methods to increase

its bioavailability and curcumin’s efficacy in influencing the functionality and improving gut

health may be associated with a concentration in the intestine due to its poor absorbability

(Lopresti, 2018).

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Turmeric is comprised of a set of three curcuminoids: curcumin (diferuloyl methane), demethoxy

curcumin, and bisdemethoxy curcumin, in addition to volatile oils (tumerone, atlantone, and

zingiberone), sugars, proteins, and resins. The Curcumin is alipophilic polyphenol this is nearly

insoluble in water however is pretty stable in the acidic ph. (Wang et al., 1997). Curcumin is the

major bioactive component of turmeric powder (Pawar et al., 2014), about 80% of the total

curcuminoids (Ashraf and Sultan, 2017). It can reach up to 6.8-7.3% of the product as reported

by Paul et al., (2016).

2.3.3 Antibacterial Property of Turmeric Powder

Turmeric and its derivatives have proven to have an antimicrobial property based on in vitro and

in vivo trials. Reduction of Escherichia coli counts was observed by Ahlawat et al. by

supplementing turmeric powder at 0.5% of the diet. A study by Nascimento et al. (2019)

reported

that 1% turmeric powder in the diet inhibits intestinal colonization of Salmonella typhimurium

in infected chicks. Necrotic enteritis induced by Clostridium perfringens causes high mortality

and problem in performance in poultry (Caly et al., 2015). Furthermore, coccidiosis caused by

protozoan is one of the predisposing factors of necrotic enteritis (Adhikari et al., 2020). A recent

study by Ali et al. (2020) reported that dietary supplementation of turmeric powder at 0.2% was

found to have an effective reduction of C. perfringens counts in the gut of broiler chicken.

Additionally, protozoan under genus Eimeria can be controlled by turmeric powder (Abbas et

al., 2010; Gogoi et al., 2019). The findings conclude that turmeric powder can be used to prevent

necrotic enteritis and other intestinal diseases. The reduction of bacterial counts might be due to

the antimicrobial properties of the active components of turmeric. In vitro trials confirmed that

10
curcumin is effective in controlling Streptococcus pyogenes, S. aureus, Acineto bacteriwoffii,

Enterococcus faecalis, Pseudomonas aeruginosa, Salmonella enteritica, E. coli, Bacillus subtilis,

and Klebsiella pneumoniae (Infante et al., 2014; Gunes et al., 2016; Adamczak et al., 2020).

Boeder et al., (2018) reported that turmeric extract with high curcumin content have better anti-

mycoplasma (M. hominis, M. capricolum, M. genitalium, and M. pneumoniae) activity.

Furthermore, M. gallisepticum which is the causative agent of chronic respiratory disease, can be

controlled by combining nanoparticles of turmeric, zedoary, and garlic extracts (Handharyani et

al., 2020). The essential oil from turmeric can inhibit the growth of E. coli, P. aeruginosa,

Bacillus cereus, Bacillus coagulans, Bacillus subtilis, and S. aureus, and Staphylococcus

epidermitis (Negi et al., 1999; Gonçalves et al., 2019; Kumar et al., 2020). Essien et al. (2015)

discussed that the turmeric essential oil’s significant antibacterial activity might be attributed to

turmerone level. However, Marliyana et al., (2019) reported that pure ar-turmerone did not have

antibacterial activity against S. aureus ATCC 25923, E. coli ATCC 25922, Klebsiella

pneumonia ATCC 13883, and P. aeruginosa ATCC 27853. They stress that the essential oil’s

antibacterial activity might be due to the compounds’ synergism instead of the pure compounds.

The antibacterial mechanisms of curcumin were reviewed by Zheng et al. (2020) and Kai et al.

(2020). The authors discussed that it destroys bacteria by inhibiting the bacteria’s quorum-

sensing system, down regulation of bacterial gene expression, inhibition of SOS-induced

responses, inhibition of cell division, and interfering protein synthesis by RNA disruption,

disruption of the cell membrane, and induction of reactive oxygen species.

2.3.4 Turmeric Powder versus Antimicrobial Growth Promoter

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Numerous studies recommending turmeric and its derivatives to use as an alternative to AGP in

broiler chickens have shown significant improvement or similar performance with that of

antibiotics. Attia et al. (2017) found significant improvement in feed conversion ratio (FCR) and

European production index of Hubbard broilers when fed with 0.1% turmeric powder from day 1

today 35 compared to control diet (no turmeric powder), a diet with 0.05% and 0.2% turmeric

powder, a diet with 0.1% mannan oligosaccharide (MOS), and diet with 50ppm oxytetracycline

(CTC). Samarasinghe et al. (2003) found no significant difference in feed intake, weight gain,

and FCR between broiler chicken fed 0.1% turmeric powder, 200ppm MOS, and 500ppm

virginiamycin. The author added, 0.2-0.3% turmeric powder improved protein and energy

utilization. Ahlawat et al. (2018) compared the broiler chickens’ performance fed different levels

(0.25%, 0.5%, 0.75%, and 1%) of turmeric powder and unnamed antibiotics. The authors

observed a significant difference in body weight gain at 0.5-0.75% and a significant difference in

FCR at 0.5%. Moreover, Abbas et al. (2010) found a similar coccidiostatic effect with that of

salinomycin sodium (0.024% of the diet) when 3% turmeric powder is added to the diet of

Eimeria tenella infected broiler chicken. The nano-curcumin of turmeric was effective in

controlling Eimeria species (Gogoi et al., 2019). The findings reported in those studies conclude

that turmeric powder can replace certain antibiotics as a growth promoter, giving better or at

least similar performance.

2.3.5 Effects of Dietary Turmeric Powder on laying performance.

The addition of 0.50 or 1% turmeric significantly increased the egg production. However, these

levels numerically increased the body weight gain and feed intake as compared to hens fed basal

diet (Nadia et al., 2008; Moorthy et al., 2009). Found that dietary supplementation of turmeric at

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1.0g/kg did not influence hen house egg production as well as hen day egg production

(Malekizadeh et al., 2011) reported that feeding of turmeric at 10.0 or 30.0g/kg did not influence

egg production of single comb white leg horn laying hens. (Sawale et al., 2009) recorded

increased egg production in laying hens fed with herb-derived mineral toxin binder production

containing Curcuma longa. (Emadi. et al., 2007) reported no significant difference in egg

production. Hens fed 1% turmeric powder had lower feed consumption which resulted in

reduction of egg production and egg mass compared with the control diet. The lower egg

production and egg mass might be related to the lower feed consumed by laying hens fed 1%

TRP. Egg production percentage was significantly affected by the treatments (P<0.05), with

layers fed 1% turmeric root showed higher egg production as compared to the control birds,

which were not fed this natural pigment. (Laganá et al., 2011) Found that inclusion of turmeric

root at level of 2% in the diet did not affect (P<0.05) egg production significantly in the birds

when compared to the basal diet. (Curvelo et al., 2009) Did not find any significant differences

in egg production when lower levels of annatto extract and turmeric were added to layer diets.

(Saraswati et al., 2013; Saraswati et al., 2013) reported that supplementation of turmeric powder,

regardless of period of administration, increased the total number of egg production until 9

months of age (P<0.05). Supplementation of turmeric powder in birds fed high protein ration did

not improve total number of egg production (P>0.05). Birds fed high carbohydrate ration and

supplemented with turmeric powder for 30 days prior to sexual maturity had 20% higher egg

production as compared to control. However, a longer period of turmeric powder

supplementation did not increase total number of egg production significantly. Egg production

was the highest in the layers fed diet with 0.5% turmeric powder and the lowest in the layers fed

the control diet (Park et al., 2012). Turmeric powder supplementation up to 4% in the ration of

13
laying hen showed a significant effect to improve egg production; the improved egg production

performance was apparently maintained by turmeric supplementation along the 3 periods of

experiment. (Bozkurt et al., 2014; Rahardja et al., 2015) reported that diet containing turmeric

powder at 5g/kg showed significant increase (P<0.05) in the egg production but at 1% level the

egg production was not affected significantly. (Hassan, 2016) reported that the dietary

supplementation of turmeric powder at 2 and 4% had no significant difference on egg production

in laying hens as compared to control group.

2.3.6 Feeding trials using Turmeric powder or its extracts, and Turmeric powder in

conjunction with other active agents to improve the health of laying hens

Like in broilers, most experiments on laying hens deal with the use of TRP to improve

biochemical parameters in blood, such as cholesterol fractions, as well as to decrease serum AST

and ALT activities and TAG concentration. In all cases, TRP has been successfully used. For

instance, Kermanshahi and Riasi (2006) indicate a decrease of 63.9, 50.2 and 63.3% for TAG,

total and LDL cholesterol levels, respectively, while HDL-cholesterol content was raised up to

15%. Similar results were obtained by many other authors (e.g., Malekizadeh et al., 2012; Riasi

et al., 2012; Arshami et al., 2013; Saraswati et al., 2013; Mirbod et al., 2017). Concerning

enzyme activities, Malekizadeh et al. (2012) found that AST and ALT activities decreased by 5.1

and 19.0%, in line with the results of Mirbod et al. (2017) and Saraswati et al. (2013).

On the other hand, TRP improves laying hen immunity, since it increases total immunoglobulins

(Ig) and IgG titers after sheep red blood cells (SRBC) injections (Arshami et al., 2013), and TRP

boosted the immune response to NDV and SRBC antigens, although feeding TRP also decreased

the heterophils : lymphocytes ratio (Mirbod et al., 2017). Another interesting effect of TRP

14
 supplementation is a decrease of E. coli counts in the ileal content of farmed laying hens (Mirbod

et al., 2017).

2.4 ORIGIN AND DESCRIPTION OF CLOVE

The symbol of dignity that is what “Clove” actually means. It is a precious and valuable spice of

the world. It is an unopened flower bud growing on a tree belonging to the family Myrtaceae

which is same as that of guavas. Cloves (Syzygium aromaticum, Eugenia aromaticum or

Eugenia caryophyllata) are the aromatic dried flower buds, which are commonly used in

biryanis, pickles, salads and garammasala. The tree that creates the miracle of nature originated

from the Moluccas Islands (Maluku islands), actually known as Spice Island (Milind and Deepa

2011). It is the common product found in the spice rack around the world. Clove buds possess

intense fragrance and burning taste. They have deep brown colour, powerful fragrant odour

which is warm, pungent, strongly sweet and slightly astringent.

Clove, is a medium sized tree (8-12m) from the Mirtaceae family and native from the Maluku

Islands in East Indonesia (Nurdjannah et al., 2012). For centuries the trade of clove and the

search of this valuable spice stimulated the economic development of this Asiatic region

(Kamatou et al., 2012). The clove tree is frequently cultivated in coastal areas at maximum

altitudes of 200m above the sea level. The production of flower buds, which is the

commercialized part of this tree, starts

15
after 4 years of plantation. Flower buds are collected in the maturation phase before flowering.

The collection could be done manually or chemically mediated using a natural phytohormone

which liberates ethylene in vegetal tissue, producing precocious maturation (Filho et al., 2013).

Nowadays, clove is cultivated in several parts of the world, including Nigeria.

Clove is rich in eugenol and is used as antibacterial in human and veterinary medicine (Rhayour

et al., 2003). About 89% of clove essential oil is eugenol (Jirovetz et al., 2006).

Clove is often referred to as the flowering buds of Syzygium aromaticum ((L.) Merr. & L.M.

Perry) or Eugenia carophyllata Thunb scientifically (Cortés-Rojas et al., 2014). Syzygium is the

largest genus of Myrtaceae family, which Clove is consists of about 1200–1800 species of

flowering plants (Cock et al., 2018). Historically, clove originated from a small island of Maluku

in Eastern Indonesia also known as the “Spice Islands” (Kamatou et al., 2012). Currently,

production of cloves is in several thousand tons in countries such as Indonesia, India, Malaysia

and SriLanka, while higher quantities are produced in the West Indies, Madagascar and

Tanzania, especially Zanzibar (Kamatou et al., 2012).

2.4.1 Phytochemical constituents of Clove

The main bioactive compound of cloves is eugenol with a concentration of about 9381.70–

14,650.00inmg/100g of fresh plant material (Cortés-Rojas et al., 2014). Cloves serve as one of

the vital leafy sources of phenolic compounds e.g. flavonoids, hydroxyl cinnamic acid, hydroxyl

benzoic acids and hydroxyl phenyl propene (Neveu et al., 2010) Phenolic compounds such as

phenolic acid and flavonoids in clove buds extracts are quercitrin (30.26mg/g), quercetin

(29.71mg/g), kaempferol (20.47mg/g), chlorogenic acid (20.13mg/g), gallic (14.07mg/g), caffeic

acid (13.98mg/g), luteolin (9.28mg/g), rutin (9.11mg/g), catechin (1.85mg/g) and ellagic acid

16
(1.61mg/g) (Adefegha et al., 2016). The presence of these phenolic compounds could possibly

enhance various bioactivities of cloves (Rubió et al., 2013; Ryu et al., 2016).

Clove buds consist of essential oil (EO) of about 15–20%, with eugenol being the major

compound (70–85%), eugenol acetate (10–15%) and β-caryophyllene (5–12%). Other minor

constituents are methylamyl ketone, gallotannic acid, ά-humulene, methyl salicylate, β humulene

crategolic acid and benzaldehyde. These constituents are accountable for the scintillating aroma

of the clove bud (Mittal et al., 2014). Clove powder showed that it consist of total soluble sugars

(32%), moisture (29.47%), crude fibre (14.37%), crude protein (6.91%), crude fat (5.86%), and

ash (5.29%) from a study by (Kaur et al., 2019). Singh et al. (2011) also reported the following

proximate composition of clove on a dry weight basis: protein (5.93%), fat (15.36%),

carbohydrates (61.93%), fiber (30.67%), ash (5.8%), and moisture (12.6%).

2.4.2 Morphology and Taxonomy of Clove

Clove is an aromatic spice tree. The term clove is taken from French word ‘clove’ and ‘clou’

which means ‘nail’. Clove is conical myrtle, medium sized tree with straight trunk which grows

up to 10 to 12m in height. The branches are semi erect, grayish in color and dense. Leaves are

large oblong to elliptic, simple obovate opposite, glabrous and possess plenty of oil glands on the

lower surface. Tree begins flowering in about 7years and continues flowering for 80years or

more.

Clove flowers are small, crimson in colour and are hermaphrodite (bisexual) borne at the

terminal ends of small branches. Each peduncle carries 3 to 4 stalked flowers and in florescence

length remains between 4-5 cm. Initially flower buds are pale yellow in color with glossy

appearance and turn green to bright red at maturity. These are 1 - 2cm long with cylindrical thick

17
ovary consisting of four fleshy sepals. Buds are divided into elongated stem and a globose

bulbous head which stimulates into nail. Commercially cloves used are air dried unopened

flower buds, 2.5cm in length and 1.25cm wide. Fruit matures nine months after flowering and the

red ovary gradually turns to reddish purple. The fruit nearly contains one or two seeds known as

‘mother of clove’. The cultivated trees are rarely allowed to reach fruit stage. These are harvested

when they develop dark red ellipsoid berry (Kamatou et al., 2012; Cortes-Rojas., et al., 2014).

Clove, Syzygium aromaticum (L.) is taxonomically classified below;

Domain - Eukaryota

Kingdom – Plantae

Sub kingdom – Viridaeplantae

Phylum - Tracheophyta

Sub phylum - Euphyllophytina

Infra phylum - Radiatopses

Class – Magnoliopsida

Sub class - Rosidae

Super order – Myrtanae

Order – Myrtales
Sub order - Myrtineae

Family - Myrtaceae

Genus - Syzygium

Specific epithet – Aromaticum (Milind and Deepa, 2011)

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2.4.3 Nutrient Content of Clove

The composition of the clove varies according to the agroclimatic conditions under which it is

grown, processed and stored. The dried clove bud contains carbohydrates, fixed oil,

steamvolatile oil, resins, tannins, proteins, cellulose, pentosans and mineral elements (Milind and

Deepa, 2011). Carbohydrates comprise about two-thirds of the weight of the spice. The dried

dark and flower buds also contain nutrients like proteins, minerals, vitamins, etc. (Milind and

Deepa, 2011).

2.4.4 Effects of Clove oil supplementation on laying performance

Mukhtar (2011) showed an increase in the feed intake by broiler chicks fed with 600mg clove

oil/kg as compared with those from the control, 200mg/kg and 400mg/kg clove oil groups, but

this increase was not significant (P>0.05). In another experiment, Hussein et al., (2019) reported

that increasing clove oil levels (0, 0.75, and 1.5mL/kg) gradually increased feed intake in

Japanese quails. Gandomani et al. (2014) reported that adding clove buds to different levels

(0.2%, and 0.4%) of laying hens and broiler does not affect daily feed consumption. Hens fed the

control diet produced less egg than birds fed the clove oil supplemented diets in this study. In

response to increasing level of clove oil supplementation, egg production linearly increased. The

addition of different levels (0.2 and 0.4%) of clove bud to laying hen feeds increased egg

production (Gandomani et al., 2014). Essential oils’ functions is based on organoleptic effect and

stimulation of organism to the production of digestive juices due to consisted compounds. As a

result, higher nutrient digestibility and absorption were observed (Arpášová et al., 2017).

2.5 SERUM BIOCHEMISTRY

19
Serum biochemistry involves the chemical analysis of substances which are embedded within the

serum such as lipids, proteins, hormones and enzymes. Testing for these various substances

provides basic knowledge about the state of the organs and tissues including the metabolic

condition of the animals (Williams, 2021).

2.5.1 Serum Proteins

Proteins are the main and most abundant constituents of the blood serum or plasma, having many

essential physiological functions. The most of proteins present in the blood are biochemically not

pure; usually, they are a mixture of simple proteins combined with other substances:

glycoproteins, lipoproteins, and other conjugated proteins (Stockham and Scott, 2002). Proteins

have a specific intramolecular structure and amphoteric nature, containing the balanced portions

of hydrophilic and hydrophobic groups (Gorinstein et al., 2002). They are macromolecules built

from one or more unbranched chains of amino acids linked by peptide bonds. The chemical

properties of the amino acids determine the biological activity of the protein (Tymchak et al.,

2010). Proteins play a central role in biological processes; some of them are involved in

structural support of connective tissues, while others play important roles in biochemical

reactions. Proteins also serve as buffers, helping in maintaining the acid-base balance and colloid

osmotic structure. Some of them act as carriers of lipids, hormones, vitamins, and minerals in the

circulatory system, and are involved in the regulation of cellular activity and immune system

(Anderson and Anderson, 2002). Other blood proteins play important roles as enzymes,

complement components, or protease inhibitors. Certain blood proteins are essential for

hemostasis and have important functions in platelet adhesion and aggregation, as well as

coagulation (Meyer and Harvey, 2004).

20
The protein constituents of the blood serum are qualitatively different from that of plasma, in

which fibrinogen has been removed by conversion into a fibrin clot together with some other

coagulation factors (Lum and Gambino, 1974; Lundblad, 2003). Although serum and plasma are

considered suitable samples for many chemistry tests, including serum total proteins, differences

in the results obtained between these 2 sample types have been reported by some authors (Miles

et al., 2004; O’Keane and Cunningham, 2006). Blood serum contains many different proteins.

Some of them are present in the blood serum in concentrations higher than mg/ml, including

albumin, immunoglobulins, haptoglobin, transferrin, and lipoproteins (Burtis and Ashwood,

2001). In addition to these major constituents, blood serum also contains many other proteins

that are secreted by cells, and tissues in very low concentrations (measured in ng/ml or pg/ml)

and in veterinary clinical biochemistry are relatively underutilized (Kennedy, 2001; Schrader and

Schulz-Knappe, 2001).

2.5.1.1 Total Protein

The total protein level in poultry serum is a measure of all the proteins in the blood, including

albumin, globulins, and other proteins. Proteins are important, as they play crucial roles in

coordinated feed intake, regulation of homeostasis and metabolism of nutrients, among others.

When ingested, proteins undergo hydrolysis in the gastro intestinal tract (GIT) and release

smaller units known as amino acids (AA) that functions in structural development, blood

components, and hormonal activities in the body (Abbasi et al., 2014). Proteins are vital in

livestock feeds and they sustain life alongside other nutrients such as carbohydrates, fats, crude

fibre, water, vitamins and minerals (Beski et al., 2015). These proteins are structural polymers,

consisting of linkages of AA to form compounds possessing carbon structure, amino and

carboxyl groups (Cheeke, 2005).

21
Several methods have been developed for the determination of total protein in serum or plasma,

which are based on different analytical methods (Zaia et al., 1998). Chemical, as well as physical

methods are available to measure total protein in biological fluids. In clinical biochemistry,

chemical methodologies are more frequently used because of the possibility to adapt these

techniques to automated analysers (Eckersall 2008). Protein is the most important solute

dissolved in serum; thus, the refractive index reflects the concentration of proteins in the sample

(George, 2001). The two major types of proteins in the blood are albumin and globulins.

Currently, the bromocresol green (BCG) and bromocresol purple (BCP) methods are the basis

for the determination of serum albumin (Watanabe et al., 2004).

2.5.1.2 Albumin

Albumin is the most abundant protein found in blood plasma or serum, and essential part of the

biochemistry profile. It is a homogenous protein fraction and is visible as a discrete zone on the

electrophoretogram. In animals, 35–50% of the total serum protein concentration is made up

from albumin (Kaneko, 1997). The shape and size of albumin fraction are very similar in all

ruminant species, which are related to its high serum concentration, homogenous electric charge,

and high staining affinity. However, there are great differences in its relative concentrations

between different animal species (Keren, 2003). Albumin can be seen on the left side of the

electrophoretogram closest to the anode, where forms a large peak (Vavricka et al., 2009).

Albumin is small size protein with a molecular weight of 69 kDa. The main functions of albumin

are the maintenance of homeostasis and transportation of substances, and it also acts as a free-

radical scavenger (Hankins, 2006). It is responsible for about 75% of the osmotic pressure of

22
plasma and is a major source of amino acids that can be utilized by the animal’s body when

necessary (Mackiewicz, 1997). It also serves as a carrier protein for many insoluble organic

substances (e.g., unconjugated bilirubin). Serum albumin is the major negative acute-phase

protein. The half-time for clearance of albumin varies from 1.9 days in the mouse to 14–16 days

in ruminants, and because of this, it may serve as a marker of chronic nutritional status (Prinsen

et al., 2004). Furthermore, may studies have established albumin as an indicator of morbidity

and mortality (Don and Kaysen, 2004).

2.5.1.3 Globulins

The globulin fractions may be found on the right side of the electrophoretogram. These peaks

include a very heterogenous group of proteins, and depending on the species, there may normally

be one or two α, one or two β, and one or two γ fractions (Kaneko, 1997). The α-fraction is the

most rapidly migrating protein of all the globulins, and in most species, it migrates as α1 (fast)

and an α2 (slow) fraction. The β-globulins belong to group of globular proteins that migrate

faster than γ-globulins in electrically charged solutions, but more slowly than α-globulins. The

main components of the β-globulin fraction are transferrin and complement, which may

correspond to the 2 subfractions (β1 and β2) identified in some animal species (Cerón et al.,

2010). The γ-globulin fraction is predominantly composed of immunoglobulins (Ig) of various

classes (IgG, IgA, IgM, IgD, and IgE). While in some animal species (cattle and goats) the γ-

globulins constitute one overall fraction, in sheep, they may be visualized as two subpeaks: the

γ1 and γ2 subfractions (Nagy et al., 2015). Immunoglobulins from the γ fraction may migrate as

fast or slow, which may be seen in these two subfractions (Kaneko, 1997).

Several factors, including non-pathological and pathological conditions, may influence the

concentrations of proteins in the serum, thus the entire profile of serum proteins (Trumel et al.,

23
1996). Many disease processes are associated with abnormal serum protein profiles. Changes in

the protein profile commonly occur as secondary symptoms in numerous diseases, but may be

also the primary symptom of some specific disease conditions (Jania and Andraszek, 2016). The

concentrations of serum proteins may be influenced also by hormonal changes and stress. Stress

may cause a decrease of serum protein and albumin concentrations, but often may be

accompanied by an increase of the α2-globulin fraction associated with the acute-phase response

(Eckersall, 2008).

2.5.2 Effects of Turmeric on Serum protein of laying hens

Kumari et al. (2007) recorded significantly higher levels of serum total protein and globulin of

broiler hens fed a diet treated with 1g/kg turmeric powder, while the serum albumin level was

lower. Al-Norii et al. (2011) found a significant increase in serum total protein concentration in

broiler chickens that received a diet containing 0.5% and 1% turmeric powder. However, Emadi

et al. (2007) suggested no significant effect on total protein and albumin concentrations in broiler

chickens treated with 0.25%, 0.5% and 0.75% turmeric powder at 21days. Ahmadi (2010)

reported that 0.3g/kg of turmeric powder had no significant effect on serum total protein,

albumin, globulin, ALT and AST enzymes compared with the control group. Similarly, there

were no significant differences in total protein, albumin and glucose in laying hens treated with

5, 15 and 25g/kg of turmeric powder (Arshami et al., 2013).

2.5.3 Effects of Clove on serum protein of laying hens

A study by Al-Yasiry & Kiczorowska, (2016) that investigated the effects of clove essential oil

on serum protein levels in broiler chickens. The study fed broiler chickens with a diet

supplemented with clove essential oil (0.05%) for 42 days. At the end of the study, the authors

24
found that the serum protein levels of the broilers in the clove oil group were significantly higher

compared to the control group. Specifically, the clove oil group had higher levels of total protein,

albumin, and globulin. The authors suggested that clove oil may have a positive effect on the

nutritional status of broiler chickens, which may lead to improved health and productivity. While

the study was conducted on broiler chickens, it is possible that similar effects of clove oil could

be observed in laying hens.

2.6 NUTRIENT DIGESTIBILITY

Nutrient digestibility is the extent to which feed nutrients are absorbed as they pass through the

bird’s digestive tract (Nhlane et al., 2020). It is the amount of feedstuff remaining in the animal

body from the total amount of feedstuff ingested by the animal. Nutrient digestibility plays

important role in nutrient utilization in animal body and thereby influences animal health and

performance. Some of the factors affecting nutrient digestibility include; Species of animal, age

of the animal, feed composition, level of feeding, frequency of feeding, processing of the feed

and the health status of the animal (Patil and Patil, 2022).

2.6.1 Effects of Dietary Clove and Turmeric Supplementation on Nutrient Digestibility

A number of studies have investigated the effects of dietary clove and turmeric supplementation

on nutrient digestibility in poultry. The supplementation of turmeric powder at 2.5, 5 and 7.5

g/kg improved the nutrient digestibility of chickens (Fawaz et al., 2022). Turmeric powder has

improved the activity of digestive enzymes and stimulated bile production (Gandhi et al., 2011;

Van Phuoc et al., 2019), which may be beneficial to enhance nutrient digestibility. Rajput et al.

(2013) reported that the addition of 200mg/kg curcumin to laying hens diet significantly

enhanced the apparent utilization of ether extract compared with control. However, Silva et al.

25
(2018) indicated that dry matter, ether extract and crude protein were not significantly different

when laying quails were fed diet supplemented with 0.5, 1.0, 1.5 and 2% turmeric powder

compared with those in control group.

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 EXPERIMENTAL SITE

The experiment was carried out at the Layers Unit, Teaching and Research Farm, LAUTECH,

Ogbomoso. Ogbomoso is located in the derived Savanna Zone that lies on longitude 4 0101East of

greenish meridian and latitude 80101 north of the equator. The latitude ranges from 300m and

600m above sea level while mean temperature and annual rainfall are 27 0C and 1247mm (Google

Earth Map, 2022).

3.2 SOURCE AND PREPARATION OF TEST INGREDIENT

26
Clove was purchased from Oja, Jagun market, Ogbomoso. Turmeric was sourced from

LAUTECH Teaching and Research Farm, Ogbomoso. Turmeric was sliced into 2mm pieces and

sundried until constant weight was achieved. The dried turmeric was ground in a mechanical

blender and sieved using a 1mm mesh to obtain turmeric powder (TP). Dried clove buds was air

dried for 24 hours prior to milling. The dried materials was pulverized using a mechanical

blender to obtain clove powder. The powdered samples was stored in a dry, clean air tight

container before they are incorporated into formulated diets.

3.3 EXPERIMENTAL DESIGN

Completely Randomized Design (CRD) was adopted for the study. A 2×2 factorial arrangement

of clove powder and turmeric powder was used to assess main and interaction effects on

measured parameters. Clove at 2 inclusion levels (0.1 and 0.2%) and turmeric at 2 inclusion

levels (0.25 and 0.5%)

3.4 FORMULATION OF EXPERIMENTAL DIET

Five experimental diets were formulated for the study. Basal diet without test ingredients served

as control. Basal diet was supplemented with 0.1% clove powder (CLP) and 0.25% turmeric

powder (TP) in T1. Basal diet supplemented with 0.2% CLP and 0.25% TP to represent T2.

Basal diet in T3 was supplemented with 0.1% CLP and 0.5% TP. Diet in T4 was basal diet

supplemented with 0.2% CLP and 0.5% TP.

Outlay of the Experiment

Control diet = Basal diet without supplementation of CLP and TP

Diet T1= Basal diet+ 0.1% CLP and 0.25% TP

27
 Diet T2= Basal diet +0.2 % CLP and 0.25% TP

Diet T3= Basal diet + 0.1% CLP and 0.5% TP

Diet T4= Basal diet + 0.2% CLP and 0.5% TP

Table 3.1: Ingredients Composition of Experimental Diets and Grower Mash

Ingredients Control T1 T2 T3 T4 Grower
mash
Maize 50.00 50.00 50.00 50.00 50.00 36.00
Maize offal 15.80 15.80 15.80 15.80 15.80 31.00
Wheat offal 3.00 3.00 3.00 3.00 3.00 17.40
SBM 18.00 18.00 18.00 18.00 18.00 10.00
Fishmeal (72% CP) 3.00 3.00 3.00 3.00 3.00 2.00
Methionine 0.10 0.10 0.10 0.10 0.10 0.10
Salt 0.25 0.25 0.25 0.25 0.25 0.25
Premixes 0.25 0.25 0.25 0.25 0.25 0.25
Bone Meal 1.80 1.80 1.80 1.80 1.80 2.00
Limestone 7.80 7.80 7.80 7.80 7.80 1.00
Clove powder - + ++ + ++ -
Ginger powder - * * ** ** -
Calc.Analysis/Total 100 100 100 100 100 100

28
Crude protein 16.97 16.97 16.97 16.97 16.97 16.45
M.E.(Kcal/Kg) 2739.90 2739.9 2739. 2739.9 2739. 2663.82
0 90 0 90
Lipids 3.31 3.31 3.31 3.31 3.31 3.36
Crude fiber 4.35 4.35 4.35 4.35 4.35 6.59
Calcium 3.62 3.62 3.62 3.62 3.62 1.26
Avail. Phosphorus 0.53 0.53 0.53 0.53 0.53 0.53
SMB= Soya bean meal += 0.1% CLP, ++ = 0.2% CLP, *0.25%TRP, ** 0.5% TRP

3.5 EXPERIMENTAL ANIMALS AND MANAGEMENT

Eighty (80) 13 weeks old, grower birds of ISA Brown strain were purchased from a reputable

commercial farm. These grower birds were offered grower mash for a period of 5 weeks prior to

the study. Thereafter, 80, 18 weeks old POL birds were randomly divided into 9 treatment

groups of 4 replicates per treatment. Each replicate contain 4 birds. All necessary medication and

vaccination were administered appropriately. A period of 12 weeks was used for the experiment.

3.6 DATA COLLECTION

Feed intake

Certain quantities of the experimental diets were offered to the birds ad libitum and feed leftover

were collected and weighed on weekly basis. The difference between the quantity of feed offered

and feed leftover was calculated and the feed intake was determined as follows:

29
 Feed intake (gram/bird/day) = Quantity supplied - Leftover

Number of birds x Number of days

Egg Production

The number of eggs laid and their weight were recorded daily using digital sensitive scale. Hen

day production was calculated using the formula below;

Hen Day Production = Number of eggs laid x100

Number of hens x Number of days

Feed Conversion Ratio

Feed conversion ratio was obtained by dividing the total feed intake with total egg weight

Feed conversion = Total Feed Intake

Total Egg weight

Feed efficiency per dozen eggs

This parameter considered the feed intake and egg production. The ratio between the feed consumed

and the number of eggs produced is indicated below;

Kg of feed consumed x 12
Total numbers of eggs laid
Feed Efficiency (per dozen eggs) =

Egg Mass (g):

To calculate egg mass, weight of the egg was multiplied by HDP

30
Egg mass = HDP X egg weight

3.7 SERUM ANALYSIS

This was carried out on the 70th day of the experiment. Two (2) laying hens from each replicate

were used to determine the serum indices. Blood samples was collected using a 5.0 ml needle

and syringe through the brachial wing vein into bottle without anticoagulant. Serum total protein

was determined using direct Biuret method as described by Lubran (1978). The serum albumin

level was determined using Bromocresol green method (Doumas and Peters, 1997). Serum

globulin was calculated as follows: Globulin = total serum proteins – serum albumin (Coles,

1986). The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST)

was determined using techniques described in Reitman and Frankel (1957). Alkaline phosphatase

(ALP) enzyme action was measured in serum, as described by Kim and Wyckoff (1991).

3.8 NUTRIENT DIGESTIBILITY

This was carried out on the 11 th week of the experiment. Two (2) laying hens from each replicate

was used to assess the utilization of experimental diets. The birds were offered 70g of the

experimental diets daily. The faecal collection was done for 3days, after 4days of pre-adjustment

to the quantity of restricted feed intake. Faecal samples was oven dried at 70 0C until constant

weight was achieved.

3.9 PROXIMATE ANALYSIS

The proximate composition of the samples of clove, turmeric and experimental diets were

determined using the procedures of AOAC (2005).

31
3.10 STATISTICAL ANALYSIS

Data collected were analyzed using One-Way Analysis of Variance of SAS (2003) software

package. The main and interaction effects of clove and ginger were assessed using 2x2 factorial

ANOVA. Duncan’s option of the same software was used to compare the means. A probability

of 5 percent was considered significant.

CHAPTER FOUR

4.0. RESULTS AND DISCUSSION

4.1 Results

The proximate composition of clove and turmeric powder is presented in Table 4.1. Clove

powder was observed to have higher crude protein (CP), crude fibre (CF) content (13.27% and

6.70% respectively) than turmeric powder (11.67% CP and 6.60% CF respectively). However,

turmeric powder had higher ash content (10.30%) than clove powder (4.60%).

The performance of laying hens fed clove and turmeric is shown in Table 4.2. Dietary treatment

significantly influenced the Average Daily Feed Intake (ADFI, P=0.006) and egg mass

(P=0.051). Hen day production (HDP), Feed conversion (FC), egg weight and feed efficiency

per dozen (FEPD) were not significantly (P>0.05) influenced by dietary treatment. The highest

(35.40 g/bird) ADFI was observed for laying hens fed diet supplemented with 0.2% clove and

0.25% turmeric (T2) while those fed control diet had the lowest ADFI. Laying hens grouped in

32
T1, T2, T3 and T4 had higher ADFI (35.08, 35.40, 35.25 and 33.78 g/hen, respectively) and egg

mass (45.24, 40.21, 42.49 and 41.53g/egg, respectively) than the control (32.03g/hen and

37.82/egg). The heaviest (45.24g) egg mass was noticed in laying hens fed diet in T2 while the

lowest (37.82g) egg mass was observed in control. The main effect of clove had significantly

effect on HDP (P= 0.048) and egg mass (P= 0.021) with hens fed diets containing 0.1% clove

had higher HDP and egg mass than those fed 0.2% clove. Furthermore, interaction effect

significantly impacted feed conversion (P=0.033) with higher inclusion level of clove (0.2%)

increased and decreased feed conversion when turmeric was included at 0.25% and 0.50%,

respectively. The best feed conversion was noticed in hens fed 0.1% clove and 0.25% turmeric

diet (T1).

Serum proteins of laying hens fed diets supplemented clove and turmeric is shown in Table 4.3.

Dietary treatment did not significantly influence serum proteins of laying hens. The main and

interaction effect of clove and turmeric had no significant effect on serum proteins of the laying

hens.

33
 Table 4.1: Proximate composition of clove and turmeric

Parameters Clove Turmeric

Crude Protein 13.27 11.68
Crude Fiber 6.7 6.6
Crude Fat 3.4 3.4
Ash 4.6 10.3
Dry matter 89.8 88.8
Moisture content 10.2 11.2

34
Table 4.2. Performance of laying hens fed clove and turmeric

Parameters Control T1 T2 T3 T4 P value SEM

Clove 0.1% 0.2% 0.1% 0.2%
Turmeric 0.25% 0.25% 0.5% 0.5%

HDP (%) 76.41 86.24 78.57 83.11 80.95 0.225 2.71

ADFI 32.03b 35.08a 35.40a 35.25a 33.78ab 0.006 0.54
(g/bird)
FC 2.55 2.31 2.63 2.47 2.42 0.232 0.09
Egg Weight 49.44 52.51 51.16 51.16 51.30 0.137 0.68
(g/egg)
Egg mass 37.82b 45.24a 40.21ab 42.49ab 41.53ab 0.051 1.41
(g/egg)
FEPD 1.51 1.45 1.61 1.52 1.49 0.388 0.05

Factorial Clove Turmeric Clove*Turmeric

ANOVA
HDP 0.048 0.870 0.240
ADFI 0.363 0.256 0.165

FC 0.099 0.786 0.033

35
Egg Weight 0.455 0.468 0.362

Egg mass 0.021 0.537 0.096

FEPD 0.184 0.570 0.070

a, b -
Means along the same row with different superscripts are significantly different (P<0.05)

HDP; Hen Day Production ; ADFI; Average daily Feed Intake ; FC; Feed Conversion ; FEPD;
Feed Efficiency Per Dozen Eggs

36
 Table 4.3. Serum proteins of laying hens fed clove and turmeric powder

Parameters Control T1 T2 T3 T4 P value SEM

Clove 0.1% 0.2% 0.1% 0.2%
Turmeric 0.25% 0.25% 0.5% 0.5%
Total protein 5.48 5.33 5.87 4.53 5.09 0.249 0.41
(g/dL)
Albumin 2.52 2.44 2.68 2.85 2.58 0.325 0.14
(g/dL)
Globulin 2.96 2.89 3.19 1.68 2.50 0.137 0.42
(g/dL)

Factorial ANOVA Clove Turmeric Clove*Turmeric

Total protein 0.23 0.90 0.98
Albumin 0.94 0.30 0.10
Globulin 0.24 0.06 0.58

37
The nutrient digestibility of laying hens fed dietary clove and turmeric is shown in Table 4.4.

Dietary treatment significantly influenced crude protein digestibility (P=0.002), ash digestibility

(P=0.0001), ether extract digestibility (P=0.048), crude fibre digestibility (P=0.0002) and

nitrogen Free Extract (NFE) digestibility (P=0.0003). Dietary treatment did not significantly

affect dry matter digestibility. The highest ash (74.22%), ether extract (92.67%) and NFE

(76.99%) digestibility were observed in laying hens fed diet supplemented with 0.1% clove and

0.5% turmeric (T3). The CP, CF, ash and EE digestibility of T1 were comparable with that of T3.

The lowest ash, ether extract and NFE digestibility were observed in hens grouped in T2, T2 and

control, respectively,

The interaction of clove and turmeric significantly influenced CP (P=0.008), CF (P=0.008) and

NFE (P=0.026) digestibility. The inclusion of clove at higher level (irrespective of turmeric

inclusion level) decreased CF and NFE digestibility. At 0.2% clove inclusion level increased CP

digestibility when turmeric was included at 0.25%, whereas at 0.2% clove (when 0.5% turmeric

was used in the diet) decreased CP digestibility.

38
 Table 4.4. Nutrient digestibility of laying hens fed clove and turmeric
Parameters Control T1 T2 T3 T4 P value SEM
Clove 0.1% 0.2% 0.1% 0.2%
Turmeric 0.25% 0.25% 0.5% 0.5%
Dry matter 70.85 74.09 72.27 75.14 72.40 0.151 1.19
c ab a ab
Crude protein 68.35 73.52 76.84 74.30 70.51bc 0.002 1.25
a a b a
Ash 73.02 70.31 63.20 74.22 71.69a 0.0001 1.24
Ether extract 92.63a 92.43a 91.27b 92.67a 91.88ab 0.048 0.34
a a b a
Crude fibre 52.21 53.59 35.83 52.64 48.41a 0.0002 2.19
NFE 65.41c 72.30b 72.25b 76.99a 71.38b 0.0003 1.26

Factorial ANOVA Clove Turmeric Clove*Turmeric

Dry matter 0.061 0.604 0.685

Crude protein 0.836 0.030 0.008

Ash 0.002 0.000 0.081

Ether extract 0.011 0.221 0.575
Crude fibre 0.000 0.018 0.008
NFE 0.024 0.105 0.026

a, b, c -
Means along the same row with different superscripts are significantly different (P<0.05)

39
4.2 Discussion

The nutritional compositions of clove bud and turmeric rhizome powder were comparable in this

study (13.27% versus 11.68% crude protein, 6.7% versus 6.6% crude fibre, 3.4% versus 3.4%

crude fat, 89.8 versus 88.8% dry matter). The proximate composition of the two herbs revealed

that these two herbs have low nutrient content (particularly CP) which could be used as

supplements without alteration in the nutrient content of the laying hens feed when included at

low dosage. Earlier studies have shown low nutritional content in turmeric and clove bud

powder. According to Adebisi et al. (2021) turmeric rhizome powder revealed 7.09 ± .04 % CP,

6.29 ± 0.01 % ash and 69.66 ± 0.01 % carbohydrate and clove bud powder contained 5.87 ±

0.02% CP, 4.95 ± 0.10% ash and 61.92 ± 0.02% carbohydrate. Ikpeama et al. (2014) also

showed that low nutritional content for turmeric rhizome powder (9.40% CP, 4.60% CF, 2.65%

ash). However, 3.4% crude fat observed in turmeric rhizome powder was lower to 6.85% crude

fat content reported by Ikpeama et al. (2014). The 13.27% CP observed for clove bud powder in

this study was significantly higher than the reports of various researchers. Suleiman et al. (2007)

reported 1.20 ± 0.02% CP and, Bello and Jimoh (2012) reported 7.00 ± 0.01% CP in clove bud

powder. Also, the 3.4% crude fat content in clove bud powder reported in this current study was

lower than the 12.1 ± 0.45% crude fat by Suleiman et al. (2007) and 18.90 ± 0.04% crude fat by

Bello and Jimoh (2012).

The high ash content (10.3%) found in turmeric rhizome powder indicated that it had a

reasonable amount of minerals. Several authors have shown that turmeric rhizome was rich in

minerals. Enemor et al. (2020) observed that the turmeric plant was high in mineral content

(38.689 ± 0.114% calcium, 19.750 ± 0.001% magnesium, 9.204 ± 0.002% potassium, 0.708 ±

0.001% iron, 7.06 ± 0.014% sodium). Adebisi et al. (2021) observed that turmeric had high

40
content of iron (40.96 ± .02%), potassium (2.489 ± 0.02%), with low content for calcium

(0.29%) and phosphorus (0.03%). Ikpeama et al. (2014) also observed that turmeric rhizome

powder had 0.21 ± 0.01% calcium and 0.63 ± 0.02% phosphorus. Feeding turmeric extracts

could be needed in maintaining strong bone, muscle contraction and relaxation, blood clotting,

reduce blood pressure, and helps in the haemoglobin formation because of its high potassium and

iron content (Latunde-Dada, 1980 and Kubinarawa et al., 2007). Imoru et al., 2018) found that

turmeric rhizome powder contained 1.67% calcium, 0.92% magnesium, 1.29% potassium, 1.07%

phosphorus and 0.06% iron. It had been suggested that presence of essential nutrients and

minerals in turmeric rhizome powder imply that it could be utilized to improve growth

performance and health status of poultry (Imoru et al., 2018). The variations could be due to soil

type, soil nutrient, farming practises, geographical locations and varied environmental conditions

(Adebisi et al., 2021).

The performance of laying hens fed diets supplemented with different levels of clove bud and

turmeric rhizome powder evaluated in this study revealed that the experimental diets

significantly increased ADFI and egg mass. The best feed conversion observed in hens fed 0.1%

clove and 0.25% turmeric diet concurred with the findings of Mohammadi et al. (2014) who

observed that birds receiving 300mg/kg clove essential oil had improved feed conversion ratio.

Dietary treatment did not significantly affect HDP and egg weight which was consistent with the

findings of Gumus et al. (2018) who reported that there were no significant differences (P>0.05)

in egg production and egg weight of hens fed turmeric-supplemented diets. The findings on

laying performance in the present study concurred with the findings of earlier studies.

Malekizadeh et al. (2012) indicated that significantly increased egg production, but egg weight

decreased insignificantly by addition of 3% turmeric rhizome powder to laying hen diets. Park et

41
al. (2012) showed that adding 0, 0.1, 0.25 or 0.5% turmeric rhizome powder into Lohmann

Brown laying hens diets from 60 to 67 weeks of age affected egg production significantly, but

not egg weight. The higher feed intake were recorded in the groups fed CBP and TRP -

supplemented diets than the control in this present study. This finding did not agree with the

observation of Devi et al. (2023) who found out that the groups fed turmeric-supplemented (0.5,

0.75 and 1.5%) diets had significantly lower feed intake than the control. Riasi et al. (2012) and

Rahardja et al. (2015) also reported lower feed intake when laying hens were fed turmeric-

supplemented diet at the rate of 1.5 to 2g and 4g/kg of feed. It was suggested that decreased feed

intake could be due to the change in aroma, palatability and the pungent smell of the turmeric.

(Devi et al., 2023). The higher turmeric powder inclusion levels in the previous studies could

account for the low feed intake of poultry when compare to the high feed intake of the present

study. Malekizadeh et al. (2012) noted that adding 1 or 3% turmeric into laying hens diet from

103 to 112 weeks of age significantly decreased feed consumption, but did not affect feed

conversion ratio, egg production, egg mass and egg weight compared with those fed diets

containing 0 and 3% turmeric powder which may imply that feeding turmeric at higher levels

could be detrimental to the performance of laying hens. The significantly increased egg mass in

the present study was in accordance with the observation of Radwan et al. (2008) which found

out that production and mass of egg increased by addition of turmeric at 0.5%. Ooi et al. (2018)

reported that 1% inclusion of turmeric in the layer diet significantly improved (P<0.05) the

overall hen-day egg production and egg weight.

The findings of clove bud powder on increased egg mass and HDP in this study were in

consistency with assertion of Gandomani et al. (2014) who found that feeding laying hens with

diet supplemented with clove bud powder (0.2 and 0.4%) increased egg production compared to

42
the control group. Hens fed diet containing 0.1% clove bud powder had higher HDP and egg

mass than those fed 0.2% clove bud powder in the present study. It showed that lower dosage of

clove bud powder was more efficient than higher CBP dose. Differences in the levels and

duration of supplementation, age and strain of birds, system of rearing, stability of the active

compounds, product source could be the reason for the variation in the effects of turmeric

supplementation on performance of poultry (Devi et al., 2023).

The non-significant effect of the two herbs on the serum protein levels of the hens in the present

study agreed with the findings of Adu et al. (2020) who indicated that 0.25% Syzygium

aromaticum leaf meal did not pose any health challenges to the laying hens, especially as it

relates to the liver (Oboh and Akindahunsi 2005). The serum protein levels in the findings of this

study did not significantly differ from the reference range of serum proteins of healthy adult

chickens (4.0 to 7.5g/dL for total protein, 2.0 to 4.0g/dL for albumin and 1.5 to 3.5g/dL for

globulin). Al-Mufarrej et al. (2019) observed that the concentration of serum total protein across

all treatments (1%, 2%, 3%, 4%, 5% and 6% clove powder, CLP) were similar at 7 and 14 days

old. However, higher levels of CLP (5% and 6% of diet) significantly reduced the concentration

of serum total protein at day 21, compared with the groups that received 1%, 2% and 3% of CLP.

The optimum nutrients digestibility for ash, ether extract and NFE were observed in laying hens

fed diet supplemented with 0.1% CBP and 0.5% TRP (T3). These particular levels of

supplementation for CBP and TRP in the experimental diet were appropriate dosage for 22 to 34

week old laying hens for nutrients digestibility. These findings were in accordance with the

findings of a number of authors. Zacharia and Ampode (2021) discovered enhanced the

digestibility of crude protein, ash and ether extract for laying quails fed diet supplemented with

5% turmeric powder. Dalal and Kosti (2018) observed that the addition of 0.5 or 1.0% turmeric

43
to hen’s diet numerically increased all nutrient digestibility coefficients compared to control

group. The bioactive components of phytogenic plants such as eugenol have been reported to be

able to stimulate digestion and enhance nutrient absorption (Reddy et al., 2004)

44
 CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATION

5.1 CONCLUSION

It could be concluded that dietary treatment did not significantly affect hen day production, egg

weight and feed conversion. However, the main effect of clove impacted significantly on HDP

with hens fed 0.1% clove had higher (P= 0.048) HDP than those fed 0.2% clove. Laying hens fed

0.1% clove and 0.25% turmeric had the highest (P= 0.051) egg mass. Clove at 0.1% inclusion

level yielded higher (P= 0.021) egg mass than those fed 0.2% clove. The best feed conversion

(P= 0.033) was observed in laying hens fed 0.1% clove and 0.25% turmeric. Laying hens fed

clove and turmeric had higher (P= 0.006) average daily feed intake than the control. The highest

ether extract (P= 0.048), ash (P= 0.0001) and NFE (P= 0.0003) digestibility was observed in

hens fed 0.1% clove and 0.5% turmeric. Higher turmeric at 0.5% depressed CP digestibility (P=

0.030) when clove was included at 0.2%.

5.2 RECOMMENDATION

It is therefore recommended that 0.1% clove and 0.25% turmeric supplemented to laying hens

diet for improved egg mass and feed conversion. It is further recommended that 0.5% turmeric

should not be supplemented in layers diet as it depressed CP digestibility.

45
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