Research in: Agricultural & Veterinary Sciences

Vol.3, No.2, 2019, pp.102-111

PERFORMANCE AND HEAMATOLOGICAL PARAMETERS OF

BROILER CHICKS GIVEN DIFFERENT LEVELS OF DRIED LEMON
GRASS (CYMBOPOGON CITRATUS) AND GARLIC
(ALLIUM SATIVUM) EXTRACT

J.O. Alagbe1*, R.A. Oluwafemi2

1
Department of Animal Nutrition, Sumitra Research Farm, Gujarat, India
2
Department of Animal Science, Faculty of Agriculture, University of Abuja, Nigeria

Abstract. A 49 day feeding trial was carried out to determine the effect of lemon grass - garlic extracts
(CLGE) on the performance and hematological parameters of broiler chickens. Two hundred and fifty
(250), day old 308 broiler chicks of mixed sex was purchased and were randomly assigned to five
treatment, each treatment was replicated 5 times consisting of 10 birds each in a completely randomized
design. Birds in treatment 1 was given 0.025g/liter of Neomycin, Treatment 2, 3, 4 and 5 were given
aqueous lemon grass-garlic extracts at 3.0ml, 6.0ml, 9.0ml and 12.0ml/liter respectively. Clean feed and
water offered ad libitum. Results showed that CLGE had significant (p<0.05) effect on final body weight,
body weight gain, daily weight gain and feed conversion ratio of the birds. Birds in T5 recorded
significantly (p<0.05) higher body weight gain and superior feed conversion ratio than the control given
0.025 g/liter Neomycin. There was also a significant (p<0.05) difference in the mortality recorded, T1 had
the highest mortality of 12%, none was recorded in T2, T3, T4 and T5. Data on hematological indices
(PCV, Hb, RBC, MCV, MCH, MCHC and WBC) revealed that CLGE significantly (p<0.05) influenced
all the parameters assessed. It was concluded that CLGE can be included up to 12ml/liters in broiler
chicken since it resulted in improved weight, feed conversion ratio without any deleterious effect on the
health and performance of the animal.

Keywords: Performance, hematological parameters, lemon grass, garlic extract, broiler chicken.
*
Corresponding Author: Dr. J.O. Alagbe, Department of Animal Nutrition, Sumitra Research Farm,
Gujarat, India, e-mail: [email protected]

Received: 31 May 2019; Accepted: 04 July 2019; Published: 12 August 2019.

1. Introduction

The side effects of synthetic drugs such as presence of antibiotic residues leads
to antibiotic resistance in humans; residues in animal tissues and the byproducts of
synthetic drug became a matter of concern in the long term usage of such drugs. This
issues have prompted the search for herbal preparations (medicinal plants) which are
safe, cheap, reduce mortality and are able to maintain the optimum growth of animals
(Phondani et al., 2010; Bentea et al., 2010). Herbal medicines are being practiced in the
form of therapy for livestock farmers because they contains phytochemicals or bioactive
chemicals which have been reported to perform multiple biological activities such as
antibacterial, antiviral, antifungal, antioxidant, antidiarrheal, anti-stress and anticancer
(Hashemi et al., 2009; Franco-Jimenez et al., 2007).
The use of plant extract or phyobiotics has also increased because of the global
awareness on the dangers of antibiotics and have been extensively used in feed as
natural growth promoters, health protectants (Hashemi et al., 2011), reduced microbial

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load and feed palatability (Mountzouris & Paraskevas, 2009). Some of these plants of
high therapeutic value includes; Azadirachtaindica, Allium sativum, Thymus vulgaris,
Curcuma longa, Piper nigrum, Cymbopogon citratus and Allium sativum.
Cymbopogon citrates (Lemon grass) belongs to the family Poaceae, is a tall
aromatic coarse grass (Burkill et al., 1996). It is found in most countries in Africa,
Europe and Asia. The plants contains steroids, alkaloids, phenols, saponin, tannins,
anthraquinines and flavonoids as phytochemical components which allows it to
functions as antibacterial, antioxidants, antifungal and antidiarrheal. According to
Thorat et al. (2017) Lemon grass leaves contains 71.03% moisture, 3.83% crude
protein, 2.94% ash, 4.76% crude fat, 9.30% crude fibre and 20.73% carbohydrate. The
leaves of the plant produces yellow or amber color and aromatic essential oil (such as
citral α, citral β, nerol geraniol, citronellal, terpinolene, geranyl acetate, myrecene and
terpinol) when squeezed (Adejuwon & Esther, 2007).
Garlic (Allium sativum) belongs to the family lily, has been used as a spice and
a native medicine for many years. It has possess excellent antibacterial, antifungal,
antiviral, antioxidant,anti-cholesteremic, anti-cancerous, and vasodilator characteristics
(Hanieh et al., 2010; Kim et al., 2009). The aqueous extract of Alliumsativum have
been shown to inhibit E.coli and Salmonella (Kumar & Berwal, 1998; Singh & Shukla,
1984). The results attributed the efficacy of garlic due to the presence alliin,
diallylsulphideand allicin (Amagase & Milner, 1993), which possess antimicrobial
activity (Tsao & Yin, 2001).
Previous studies have shown that lemon grass positively influenced the
performance and could be used as a botanical alternative in the diet of broilers (Mukhtar
et al., 2012; Thayalini et al., 2011), quails (Sariözkan et al., 2018). Similarly, Ozougwu
et al (2011) reported a significant difference (p<0.05) in blood parameters white albino
rats orally fed 300mg/kg garlic extract. These plant extracts have been found to be
loaded with several bioactive chemicals, therefore a synergistic combination of this
plants will give a positive result on the performance of animals, save cost and ensure
total safety in the final products obtained.
Therefore, this experiment was designed to evaluate the performance and
hematological parameters of broiler chicks fed different levels of dried lemon grass
(Cymbopogon citratus) and garlic (Allium sativum) extract mixture.

2. Materials and methods

Site of the experiment

The experiment was carried out at Division of Animal Nutrition, Sumitra
Research Farm, Gujarat, India during the month of May to July, 2018.

Collection and sample preparation

Fresh, mature and healthy leaves of Cymbopogon citratus were obtained within
the farm premises,the leaves were thoroughly washed with running tap water to remove
the debris and allowed to dry under shade for 11 days until a constant weight was
obtained, The dried samples was blended into fine powder using an electric blender and
stored in air tight container. 20.00 grams of Cymbopogon citratus powder was mixed
with 100ml distilled water for 48 hours after which it was filtered using Whatmann
filter paper to obtain Cymbopogon citratus extract (CCL).

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Garlic cloves were purchased from a local market in Gutpa, the cloves were
carefully peeled, cut into pieces and dried for 7 days. The dried garlic was blended into
powder with an electric powder, extracts were prepared by soaking 100g dried garlic
powder in one liter of water for 24 hours. The next morning, the extract (GLE) was
obtained by filtering the infusion using a filter paper.
CCL and GLE were mixed in another clean bowl in the ratio of 1:1 to obtain
lemon grass/garlic extract mixture (CLGE). The powdered samples and experimental
diet were also subjected to proximate analysis to determine the crude protein, ether
extract, crude fibre and ash according to AOAC (1990).
Phytochemical components of the extracts were determined according to
methods described by Harbone (1973); Odebiyi & Sofowora (1978).

Pre-experimental activities
The house was cleaned and well disinfected before the commencement of the
experiment, wood shaves were also spread on the floor of the pen.

Animal management and experimental diets

A total number of two hundred and fifty (250) one day commercial unsexed
broilers of Ross-308 strain were obtained from a commercial hatchery in India and
transported to the farm. The chicks were weighed individually at the beginning of the
experiment. They were wing – banded and distributed randomly into 5 treatments of
250 chicks. The birds were kept under similar conditions of management throughout the
experimental period. Chicks were bought vaccinated according to the prevailing disease
condition in the environment. Water soluble multi-vitamin (Miavit Super) was given to
the chicks before 3 days of vaccination and 3 days after vaccinations in order to guard
stress. Each pen was equipped with feeders and drinkers to allow ad libitum
consumption of feed and water. Light was provided approximately 24 hours in a form of
natural light during the day and artificial light during the night.
The chicks were fed on 5 dietary treatments. The first group A was given
0.025g/litre Neomycin in water. The other groups B, C, D and E were given CLGE at
levels 3.0ml/litre, 6.0ml/litre, 9.0ml/litre and 12.0 ml/ litre of water respectively. The
basal diet was formulated to meet the nutrients requirements of broiler chicks according
to the (NRC, 1994).

Experimental design and parameters measured

Experimental design used was a completely randomized design. Daily feed
intake (g) was calculated by difference between feed offered and the left over, feed
conversion ratio was determined as feed intake divided by body weight gain, water
consumption and mortality were recorded daily throughout the experimental period.
Mortality were recorded daily and all management practices were strictly observed
throughout the experimental period.

Blood sample analysis

At the 7th week of the experiment, blood samples were collected from three
randomly selected birds per replicate, the samples were collected via the branchial vein
to aspire 5mls of blood from each bird into bottles containing Ethylene Diamine Tetra
Acetate (EDTA) for haematological analysis. Some of the haematological parameters
measured are Pack cell volume (PCV), Red blood cell (RBC), White blood cell (WBC),

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Haemoglobin concentration (Hb) and absolute counts of neutrophils, lymphocytes,

monocytes and eosinophils which were all computed according to standard techniques
as reported by Jain (1986).
MCV, MCH and MCHC were calculated according to Jain (1986) method as
shown below:
MCV (fl) = PCV (%) ×10/RBC (106/µL)
MCH (pg) = Hb (g/dl) × 10/RBC (106/µL)
MCHC (%) = Hb (g/dl) × 100/PCV (%)

Statistical analysis
All data generated were subjected to a one way analysis of variance (ANOVA)
and treatment means were compared using Steel and Torrie (1986). Differences among
treatment means were separated by least significance difference test.

Table 1. Composition of experimental diets

Ingredients 0-4 weeks (Starter)Kg 4-7weeks (Finisher)Kg
Maize 52.00 60.99
Soya meal 38.60 30.60
Groundnut cake 3.00 3.00
Fish meal (72%) 1.00 0.00
Bone meal 3.00 3.00
Limestone 1.50 1.50
Lysine 0.15 0.15
Methionine 0.20 0.20
Toxin binder 0.01 0.01
Premix 0.25 0.25
Salt 0.30 0.30
Total 100.0 100.0
Determined analysis
ME (Kcal/kg) 2990.9 3110.4
Crude protein (%) 23.23 20.51
Ether extract (%) 3.67 3.01
Crude fibre (%) 3.79 4.08
Calcium (%) 1.11 1.08
Phosphorus (%) 0.87 0.95

* Premix supplied per kg diet :- Vit A, 10,000 I.U; Vit E, 5mg; Vit D3, 3000I.U, Vit K, 3mg; Vit B2,
5.5mg; Niacin, 25mg ; Vit B12, 16mg ; Choline chloride, 120mg ; Mn, 5.2mg ; Zn, 25mg ; Cu, 2.6g ;
Folic acid, 2mg ; Fe, 5g ; Pantothenic acid, 10mg ; Biotin, 30.5g ; Antioxidant, 56mg

Table 2. Proximate composition of dried lemon grass leaf meal (CCL) and Garlic powder (GRE)

Parameters % lemon grass powder % Garlic powder

Dry matter 90.45 93.11
Crude protein 13.71 4.60
Crude fibre 9.76 6.11
Ether extract 2.51 0.66
Ash 8.77 5.11
Nitrogen free extract (NFE) 65.25 83.52

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Table 3. Phytochemical components of CCL and GRE

Components (mg) CCL GRE Safe recommended levels (mg)

Tannins 0.650 7.00 15.0
Alkaloids 0.520 4.59 3.50
Saponins 0.700 - 7.02
Phytate 0.001 27.80 23.40
Flavonoids 0.532 33.03 -
Oxalate - 1.02 1.3-1.8
Phenol 0.400 0.041

Table 4. Effect of feeding different levels CLGE on the performance of broiler chicken

Parameters Treatments
Breed (Ross 308) 1 2 3 4 5 SEM
No of birds 50 50 50 50 50 -
Live body weight (g)
Initial weight 42.10 40.01 40.03 40.08 40.71 0.08*
7th day 110.5 110.2 110.0 110.4 111.0 3.51*
28th day 889.3 845.1 850.6 856.8 860.0 7.20*
49th day 1775.0c 2001.7b 2088.2a 2113.0a 2190.6a 30.21*
Body weight gain (g)
0-7 days 71.22 70.07 70.01 69.98 71.33 1.58*
7-28 days 779.0 735.0 742.1 740.7 743.5 5.02*
0-49 days 1534.7c 1961.7b 2048.2b 2072.9a 2081.8a 33.18*
Feed intake (g/bird)
0-7 days 1413.00 1409.31 1401.11 1400.23 1400.08 12.34*
7-28 days 2140.22 2100.90 2100.08 2100.03 2100.00 22.56*
0-49 days 3640.88 3633.07 3624.01 3602.08 3600.01 35.05*
Feed conversion ratio (feed/gain)
7-28 days 2.41a 2.49a 2.47b 2.46b 2.46c 0.07*
0-49 days 1.99a 1.78a 1.73b 1.71b 1.70c 0.04*
Mortality
6/50 0/50 0/50 0/50 0/50 -
% Mortality 12.00 - - - - -

abc
means different superscript along rows differs significantly at p<0.05
*: Significant difference

Table 5. The effect of feeding different levels CLGE on the haematological traits of broiler chickens

Parameters Treatment
1 2 3 4 5 SEM
PCV (%) 30.10c 37.43b 39.56 b
39.71a 39.96a 0.04*
Hb (g/dl) 9.88c 11.01c 12.06 b
13.71b 13.80a 0.27*
RBC ×106/µL 2.01c 3.61b 3.68 b
3.71a 3.95a 0.01*
MCV (fl) 149.8a 103.7c 107.5 b
107.0b 101.2c 4.03*
MCH (pg) 49.15a 30.50c 32.77 c
36.95b 34.94b 0.07*
MCHC (%) 32.82c 29.41b 30.49b 34.53a 34.51a 0.03*
WBC ×106/µL 19.89c 22.56b 25.35 b
28.05a 31.22a 0.12*

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Differentials (106/µL)
Lymphocytes 14.18c 15.90c 16.56b 17.08a 18.22a 0.20*
Monocytes 0.99c 1.44b 1.33b 1.35b 1.38a 0.05*
Heterophils 5.09c 6.06b 6.17c 6.45b 7.58a 0.22*
Basophils 1.04c 2.10c 2.17b 2.23b 2.90a 0.05*
Eosinophils 1.03c 1.41b 1.51a 1.58a 1.74a 0.02*

abc
means different superscript along rows differs significantly at p<0.05
*: Significant difference

3. Result and discussion

Table 1 shows the chemical composition of the experimental diets. The starter
diet contained 23.23% crude protein, 3.67% ether extract, 3.79% crude fibre, 0.87%
phosphorus, 1.11% calcium and 3000.9 MEkcal/kg energy while the finisher diet
contained 20.51% crude protein, 3.01% ether extract, 4.08% crude fibre, 1.08%
calcium, 0.95% phosphorus and 3110.4 MEkcal/kg energy. The basal diet was
formulated to meet the nutrients requirements of broiler chicks according to the (NRC,
1994).
The proximate components of lemon grass leaf meal and garlic powder is
revealed in Table 2. The proximate components of lemon grass leaf meal are 90.45%,
13.71%, 9.76%, 2.51%, 8.77% and 65.25% for dry matter, crude protein, crude fibre,
ether extract, ash and nitrogen free extract. The values obtained is parallel with the
findings of Sariözkan et al. (2018). Garlic powder contained dry matter (93.11%), crude
protein (4.60%), crude fibre (6.11%), ether extract (0.66%), ash (5.11%) and nitrogen
free extract (83.52%). The result obtained during this study is in agreement with the
findings of Ayoola et al. (2016).
Results on the phytochemical analysis of Lemon grass leaf extract and garlic
extract revealed the presence of tannin (0.650mg/100g), alkaloids (0.520mg/100g),
saponins (0.700mg/100g), phytate (0.001mg/100g), flavonoids (0.532mg/100g) and
phenol (0.400mg/100g) while those of garlic powder are tannin (7.00mg), alkaloid
(4.59mg), phytate (27.80mg), flavonoids (30.03mg), phenol (0.041mg) and oxalate
(1.02mg) respectively. Lemon grass leaf extract had a higher level of saponin followed
by tannin, flavonoids, alkaloids, phenol and phytate respectively as presented in Table
3. This is consistent with the finding of Asaolu et al. (2009), though a slightly lower
value was recorded for tannin (0.600mg), this may be attributed to differences in the
variety, environmental condition and stage of growth of the plant. However, all values
are below the lethal dose reported by Kumar et al. (2010); European Food Safety
Authority (2014) on scientific opinion on the safety and efficacy of tannic acid when
used in feed for all animal species. According to EFSA (2010), the use of tannic acid as
feed additive up to 15mg/kg is safe for all animal species. Garlic extract had a higher
level of flavonoid (30.03mg) when compared with the other parameters. Similar
observation was made by Vaghasiya et al. (2011) who reported that Aristolochia
bracteolate contained a higher percentage of flavonoid during the phytochemical
analysis of some medicinal plants from Western region of India.
Saponin are play an active role as anti-inflammatory and immune stimulating
remedies (Cheeke et al., 2006). Tannins are been reported to perform antibacterial and

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antiviral activity (Enzo et al., 2007). Phenol are antioxidants which prevents the
oxidative damage of molecules such as DNA and protein (Ojewuyi et al., 2014).
The performance of broiler fed different levels of CLGE is presented in Table 4.
The average initial body weight was similar along treatments. However, the final
average weight was higher (P<0.05) for birds in T5 (2190.6g), T4 (2113.0g) and T3
(2088.2g) than those in T2 (2001.7g) and T1(1775.0g). According to Preston & Leng
(1987), the growth rate of an animal is determined by the feed intake and digestibility
with feed intake being determined by nutrients. The result of the current study was also
consistent with those reported for broilers, when supplemented with 1250 ppm Citrus
sinensis peel extract (Ebrahimi et al., 2014). Similarly, Alabi et al. (2017) reported that
broilers fed 90ml/liter aqueous Moringa leaf extract had the highest body weight and
growth rate (2392.00g and 53.61g respectively). Conversely, Hassan et al. (2014)
reported a higher feed intake and lower body weight gain in broiler chicken fed diets
without microbials.
Feed conversion ratio was significantly different (P<0.05) among the treatments.
The best feed conversion ratio (FCR) was observed in birds fed 40ml/liter of CLGE.
This was similar with the finding of Nurani et al. (2016) who noted that addition of
15ppm marigold leaf extract in the diet of quails increased their feed conversion ratio.
Mortality rate was also affected by the inclusion of CLGE at 40ml/liter in T5. 12%
mortality was recorded in birds given 10ml/liter (T1), none was recorded in T2, T3, T4
and T5 respectively.
Table 5 shows the effect of feeding different levels CLGE on the haematological
traits of broiler chicken. The PCV, Hb, RBC, MCV, MCH and MCHC values ranged
31.10 – 39.96%,9.88-13.80g/dl, 2.01 – 3.95 106/µL, 101.2 – 149.8 fl, 34.94 – 49.15 pg,
29.41 – 34.53 % respectively while the WBC values ranged 19.89 - 31.22 106/µL. There
was a significant (P<0.05) among the treatments. This was similar with the finding of
Ozougwu (2011) who noted that administration of garlic extract at 300mg/kg showed
positive haematological activities in rats but contrary to the reports of Oleforuh-Okoleh
et al. (2015) when banana leaf was used as a phytoadditive in the diets of broiler chicks.
Similarly, Alagbe (2019) also reported that dried Centella asiatica leaf meal did not
(P>0.05) significantly affect the haematological parameters of broilers when fed at
6%.However, all the values of the haematological parameters fall within the ranges for
broilers as reported by Talebi et al. (2005); Albokhadaim (2012); Al-Nedawi (2018).
Hematological studies have been found useful for disease prognosis and for therapeutic
and stress monitoring (Braun et al, 2010) but can vary due to age, gender, environment,
infection and poisoning (Khan & Zafar, 2005). Red blood cell is involved in the
transport of oxygen and carbon dioxide in the body (Isaac et al, 2013). This is a clear
indication that birds in T5 will have a have enough oxygen especially in situation of
oxygen starvation. Onu & Aniebo (2011) reported that hematocrit or PCV is an index of
toxicity, lower value could be a sign of anemia. WBC helps to fight against infections
and provide resistance against diseases (Soetan et al., 2013).

4. Conclusion

The growth and hematological parameters measured showed significant

differences. It could therefore be concluded that CLGE could be efficiently utilized and
tolerated by broiler chickens up to 40ml/liter inclusion level without any negative effect

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on the performance and health status of broiler chickens. CLGE it is safe, efficient and
should be encouraged among small and medium scale farmers.

Recommendation
Further research needs to be carried to determine the effect of CLGE on bacteria
load count, to ascertain whether it can reduce the load of pathogenic or opportunistic
bacteria in the gut of birds.

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