Nutrición Animal Tropical 19 (2): 38-62. Julio-Diciembre, 2025
ISSN: 2215-3527 / DOI: 10.15517/6ebwzj88
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1Department of Animal Production and Health, The Federal University of Technology, Akure, Nigeria. Email address:
aoayeni@futa.edu.ng (https://orcid.org/0000-0001-7558-0781).
2Department of Animal Production and Health, The Federal University of Technology, Akure, Nigeria. Corresponding author:
popoolaaph171714@futa.edu.ng (https://orcid.org/0009-0005-0551-008X).
3Department of Animal Production and Health, The Federal University of Technology, Akure, Nigeria. Email address:
joagbede@futa.edu.ng (https://orcid.org/0000-0001-7297-6350).
Recibido: 17 julio 2024 Aceptado: 01 octubre 2025
Esta obra está bajo licencia internacional CreativeCommons Reconocimiento-NoComercial-SinObrasDerivadas 4.0.
Technical note
Impact of selected phytogenic feed additives on performance, haematological,
biochemical, and antioxidant enzyme responses in 64-week-old laying hens
Akinlolu Oluwafemi-Ayeni
1, Oluwatobiloba Emmanuel-Popoola
2 , Johnson Oluwasola-Agbede
3
ABSTRACT
The effects of turmeric, ginger, and garlic powders on the performance, blood profile, serum
biochemistry, and antioxidant properties of 64-week-old laying hens were evaluated. The
study was carried out at the Teaching and Research Farm of the Department of Animal
Production and Health, The Federal University of Technology, Akure (FUTA), Ondo State,
Nigeria. A 12-week feeding trial was conducted with a total of 135 Isa Brown laying hens,
arranged in a completely randomized design. Five diets were formulated for the prosecution:
Diet 1 (control); Diet 2 (3.00% addition of turmeric); Diet 3 (3.00% addition of ginger); Diet 4
(turmeric and garlic at 2.25% and 0.75%, respectively); and Diet 5 (ginger and garlic at 2.25%
and 0.75%, respectively). The proximate compositions of the phytogenic feed additives (PFAs)
were determined as follows: moisture (5.03 – 7.50%); ash (3.26 – 6.11%); ether extract (7.39 –
14.58%); crude fiber (10.28 – 13.79%); crude protein (3.07 – 7.23%); and nitrogen-free extract
(58.88 – 61.54%). The performance indices were not significantly influenced by the test diets
(p> 0.05), except for hen-day production (HDP). The lowest HDP (57.50%) and feed conversion
ratio (FCR) (2.37%) were observed in Diet 4. No significant differences were recorded among
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39
the haematological parameters and biochemical indices (p > 0.05). Glutathione peroxidase
(GSH) activity was significantly highest (2.02 μM/mL) in birds fed Diet 2 (p < 0.05). The activities
of catalase (0.80 μM/mg protein) and superoxide dismutase (60%) were significantly (p < 0.05)
highest in birds on Diet 5. The results of this study indicate that PFAs do not exert adverse
effects on the health of older laying hens; on the contrary, they help maintain productive
performance and improve serum and antioxidant profiles.
Keywords: Old-laying hens, turmeric, ginger, garlic, phytogenic feed additives.
RESUMEN
Impacto de aditivos fitogénicos seleccionados sobre el rendimiento, la respuesta
hematológica, bioquímica y antioxidante en gallinas ponedoras de 64 semanas de edad. Se
evaluaron los efectos de la cúrcuma, el jengibre y el ajo en polvo sobre el rendimiento, el perfil
sanguíneo, el suero y las propiedades antioxidantes de gallinas ponedoras de 64 semanas de
edad. El estudio se llevó a cabo en la Granja de Enseñanza e Investigación del Departamento
de Producción y Salud Animal de la Universidad Federal de Tecnología, Akure (FUTA), Estado
de Ondo, Nigeria. Se realizó un ensayo de alimentación de 12 semanas con un total de 135
gallinas ponedoras de la cepa Isa Brown, dispuestas en un diseño completamente aleatorio.
Se formularon cinco dietas para el ensayo: Dieta 1 (control); Dieta 2 (3% de cúrcuma); Dieta 3
(3% de jengibre); Dieta 4 (cúrcuma y ajo al 2,25% y 0,75%, respectivamente); y Dieta 5 (jengibre
y ajo al 2,25% y 0,75%, respectivamente). La composición proximal de los aditivos fitogénicos
para piensos (AFP) se determinó de la siguiente manera: humedad (5,03 – 7,50%); cenizas (3,26
– 6,11%); extracto etéreo (7,39 – 14,58%); fibra cruda (10,28 – 13,79%); proteína cruda (3,07 –
7,23%); y extracto libre de nitrógeno (58,88 – 61,54%). Los índices de rendimiento no fueron
significativamente influenciados por las dietas de prueba (p > 0.05), excepto la producción
diaria por gallina (HDP). Los valores más bajos de HDP (57,50%) y de relación de conversión
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alimentaria (FCR) (2,37%) se obtuvieron en la Dieta 4. No se observaron diferencias
significativas entre los parámetros hematológicos ni entre los índices bioquímicos evaluados
(p > 0.05). La actividad de la glutatión peroxidasa (GSH) fue significativamente mayor (2,02
uM/mL) en las aves alimentadas con la Dieta 2 (p < 0.05). Las actividades de catalasa (0,80
uM/mg proteína) y superóxido dismutasa (60%) fueron significativamente más altas (p < 0.05)
en las aves alimentadas con la Dieta 5. Los resultados de este estudio indican que los AFP no
generan efectos adversos sobre la salud de las gallinas ponedoras de mayor edad; por el
contrario, contribuyen a mantener el rendimiento productivo y a mejorar los perfiles séricos y
antioxidantes.
Palabras clave: gallinas ponedoras senescentes, cúrcuma, jengibre, ajo, aditivos alimentarios
fitogénicos.
INTRODUCTION
Livestock production plays a vital role in the agricultural economies of developing countries,
providing not only a direct source of food but also essential raw materials such as hides, wool,
fertilizer, and fuel (Okoro, 2016). Within this sector, poultry production is particularly significant
but faces a major challenge in nutrition, as feed accounts for approximately 70% of total
production costs (Mmadubuike and Ekenyem, 2001). Therefore, improving feed efficiency while
reducing costs is crucial to ensure the sustainability of poultry farming (Alhotan, 2021).
In the past, growth-promoting antibiotics were commonly used as feed additives. However,
concerns about issues such as disruption of the natural gut microbiota and the development
of antibiotic resistance in both bacteria and humans have led to their restriction or ban in many
countries (Borazjanizadeh et al., 2011; Diarra et al., 2011). As a result, the use of herbs and
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medicinal plants, such as turmeric, ginger, and garlic, as natural alternatives to antibiotics has
gained momentum in the last decade, owing to their wide range of potential health benefits
(Ali et al., 2008; Mahesh and Prabhakar, 2018).
Turmeric rhizome (
Curcuma longa
), a member of the ginger family (Zingiberaceae), is a widely
recognized medicinal herb (Barazesh et al., 2013). According to Rajesh and Devvrat (2018),
turmeric contains several active compounds, including curcuminoids, aromatic turmerones
(32.5%), α-turmerones (15.6%), β-turmerones (17.1%), and curlone. Curcuminoids have been
shown to exhibit a broad spectrum of biological activities, including antioxidant, antibacterial,
antifungal, antiprotozoal, antiviral, anticoccidial, and anti-inflammatory effects (Rajesh and
Devvrat, 2018).
Ginger, the rhizome of
Zingiber officinale
, is widely used as a delicacy, spice, and in traditional
medicine (Okoro, 2016). Previous research has identified nine compounds in ginger capable of
binding to serotonin receptors, potentially influencing gastrointestinal function. Ginger
oleoresins have been associated with diverse pharmacological properties, including
antimicrobial, anti-inflammatory, antioxidant, anti-hypercholesterolemic, anti-hyperglycemic,
and antispasmodic effects (Banerjee, 2018). In addition, ginger extract may regulate free radical
levels and lipid peroxidation, and it has demonstrated antidiabetic potential (Al-Amin et al.,
2006; Morakinyo et al., 2011).
Garlic (
Allium sativum
) is widely used both as a food flavoring agent and as a traditional
remedy for various ailments (Sallam et al., 2004; Hanieh et al., 2010). Animal studies have
demonstrated that garlic exhibits hypolipidemic, hypotensive, hypoglycemic, antithrombotic,
and antiatherogenic effects. Its bioactive compounds, primarily flavonoids and organosulfur
derivatives, have also shown therapeutic and antioxidant properties in laying birds (Navidshad
et al., 2018).
Blood biochemical parameters and antioxidant activity are critical indicators of an organism’s
health status and nutrient metabolism (Lokesh et al., 2012). Recent studies investigating the
use of turmeric, ginger, and garlic as additives have reported promising outcomes, including
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improved weight gain, enhanced feed efficiency, reduced mortality, and increased survivability
in poultry (Issa and Omar, 2012; Oleforuh-Okoleh, 2014). Therefore, the present study aimed
to evaluate the beneficial effects of turmeric, ginger, and garlic powder on the performance,
hematological profile, serum biochemistry, and antioxidant capacity of aging laying hens.
MATERIALS AND METHODS
Experimental Site
The field study was conducted at the Teaching and Research Farm of the Department of
Animal Production and Health at The Federal University of Technology, Akure (FUTA), Ondo
State, Nigeria.
Collection, processing of garlic, ginger, and turmeric, and proximate composition analysis
Fresh turmeric, garlic, and ginger were purchased, washed to remove dirt, and peeled before
being chopped into smaller pieces. The turmeric and ginger rhizomes were air-dried at room
temperature (28 °C) for five days, milled into a fine powder, and stored in airtight containers.
Garlic bulbs were peeled and oven-dried at 125 °C for the first two hours, then adjusted to 102
°C for four hours before milling. The additives were incorporated into the experimental diets
in their dehydrated powdered form at the specified inclusion levels (3.00% turmeric, 3.00%
ginger, 2.25% turmeric + 0.75% garlic, and 2.25% ginger + 0.75% garlic). No extracts or
essential oils were used in this study.
The proximate composition (moisture, ash, crude fiber, and ether extract) was determined
using AOAC (2009) methods. Crude protein content was determined using the micro-Kjeldahl
method as described by Agbede et al.
(2009), while nitrogen-free extract (NFE) was calculated
algebraically using the formula provided by AOAC (2009):
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%NFE = [100 – (%Moisture + %Ash + %Crude Protein + %Ether Extract + %Crude Fiber)]
Experimental layout and hens’ management
A 12-week feeding trial arranged in a completely randomized design was conducted to
evaluate the performance of hens fed diets containing garlic, ginger, and turmeric. One
hundred thirty-five, 64-week-old, Isa Brown laying birds were obtained from the poultry
section of the Teaching and Research Farm of the Department of Animal Production and
Health at the University. The birds were housed in deep-litter cages in an open-sided house
and divided into five dietary groups, each consisting of three replicates of nine birds. The
control diet was formulated to meet the NRC (1994) requirements for laying hens, while the
other diets were duplicates of the control with various phytogenic feed additives (PFAs).
Accordingly, the five diets used for the trial, as presented in Table 1, were: Diet 1, control diet
with no inclusion of PFAs; Diet 2, 3% turmeric; Diet 3, 3% ginger; Diet 4, 2.25% turmeric +
0.75% garlic; and Diet 5, 2.25% ginger + 0.75% garlic. The birds were fed their respective
experimental diets and provided water
ad libitum
throughout the experiment.
The response criteria included feed intake (FI), feed conversion ratio (FCR), and hen-day
production (HDP). Daily FI was calculated as the difference between the weight (g) of the feed
administered and the weight of the leftover feed. The individual body weight of all hens was
recorded at the start of the first phase and at the end of the last phase, using a Kenny Camry
electronic price platform scale, to determine the average weight gain. Egg production was
recorded daily for each treatment, and the average egg production rate (hen-day percentage)
was calculated for each phase.
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Table 1. Gross composition of diets fed at laying phase.
Ingredients
Diet 1 (kg)
Diet 2 (kg)
Diet 3 (kg)
Diet 4 (kg)
Diet 5 (kg)
Turmeric
0.00
3.00
0.00
2.50
0.00
Garlic
0.00
0.00
0.00
0.50
0.50
Ginger
0.00
0.00
3.00
0.00
2.50
Corn
59.50
56.50
56.50
56.50
56.50
GNC*
14.00
14.00
14.00
14.00
14.00
Soybean meal
8.00
8.00
8.00
8.00
8.00
Wheat offal
6.00
6.00
6.00
6.00
6.00
DCP*
2.00
2.00
2.00
2.00
2.00
Limestone
7.00
7.00
7.00
7.00
7.00
Premix
0.30
0.30
0.30
0.30
0.30
Methionine
0.10
0.10
0.10
0.10
0.10
Lysine
0.30
0.30
0.30
0.30
0.30
Salt
0.30
0.30
0.30
0.30
0.30
Vegetable oil
2.50
2.50
2.50
2.50
2.50
*GNC = Groundnut Cake; DCP = Dicalcium phosphate.
Data Collection
Haematological And Serum Biochemical Studies
Three birds were randomly selected from each replicate, and 10 mL of blood was collected
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through their jugular veins. Of this, 3 mL of blood was drawn into an EDTA plastic tube for
hematological analysis. Packed cell volume (PCV), haemoglobin (Hb) concentration, red blood
cell (RBC) and white blood cell (WBC) counts, mean corpuscular haemoglobin (MCH), mean
corpuscular volume (MCV), mean corpuscular haemoglobin concentration (MCHC),
lymphocytes, and monocytes were measured following methods described by Dacie and Lewis
(1991). BIOBASE automatic biochemistry analyser was used to assess serum creatinine,
aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol, albumin,
globulin, and glucose.
Antioxidant assay
Serum glutathione peroxidase (GSH-Px) activity was measured according to the method of
Paglia and Valentine (1967), using a Randox/RS-504 kit (Ransel, Randox Laboratories, UK).
Superoxide dismutase (SOD) activity was determined using the Ransod enzyme kit
(RANDOX/SD-125) following the procedure by Nishikimi et al. (1972). Malondialdehyde (MDA)
concentration was measured with the Elabscience MDA colorimetric kit in accordance with the
method by Ohkawa et al. (1979). Total antioxidant capacity (T-AOC) was assessed using auto-
analyzers from BioMed Diagnostics (Egypt) following the method described by Koracevic et al.
(2001).
Statistical analysis
All data were subjected to one-way analysis of variance (ANOVA) using IBM SPSS Statistics
version 23.0 (IBM Corp, 2015). When significant differences were detected, Duncan´s multiple
range test (DMRT) was applied to separate the means.
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Below is the statistical model:
Yij = U + Ti + Eij
Where:
Yij is the individual observation of all treatments.
U is the universal means.
Ti is the effect of the diets, i.e., treatment.
Eij is the experimental error.
RESULTS
Proximate composition
The proximate composition of the phytogenic feed additives (PFAs) used in the experimental
diets varied across treatments, as shown in Table 2. Diet 4 exhibited the highest values for
moisture, ash, and crude fiber, whereas Diet 2 showed the highest fat and extract contents.
Crude protein levels were highest in Diet 3 and lowest in Diet 5. Significant differences (p <
0.05) were observed among the diets for all measured parameters, indicating distinct
compositional profiles among the PFAs.
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Table 2. Proximate composition of phytogenic feed additives
Nutrients (%)
Diet 2
Diet 3
Diet 4
Diet 5
±SEM*
P value
Moisture
5.03d
7.08b
7.50a
5.89c
0.01
0.001
Ash
3.26d
5.69b
6.11a
4.16c
0.01
0.001
EE*
14.98a
9.61c
7.39d
12.50b
0.06
0.001
CF*
11.52c
10.28d
13.79a
13.49b
0.03
0.001
CP*
3.67c
7.23a
6.33b
3.07d
0.07
0.001
NFE*
61.54a
60.11b
58.88c
60.89a
0.12
0.001
a,b,c – Means on the same row having different superscripts are significantly (p < 0.001) different. *SEM
= standard error of the mean; EE = ether extract; CF = crude fiber; CP = crude protein; NFE = nitrogen
free extract.
Performance indices
The performance indices (Table 3) were not significantly (p ˃ 0.05) influenced by the
experimental diets, with the exception of HDP. The highest HDP was observed in birds fed Diet
3 and 5; whereas the lowest HDP was recorded in birds fed Diet 4.
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Table 3. Performance indices of laying birds fed diet containing phytogenic feed additives.
Diet 1
Diet 2
Diet 3
Diet 4
Diet 5
SEM*
P-value
Initial weight (kg)
1.84
1.97
1.77
1.87
1.81
0.02
0.090
Final weight (kg)
1.99
2.02
1.93
1.92
1.95
0.02
0.590
Weight gained (kg)
0.15
0.05
0.16
0.04
0.14
0.02
0.100
FI/hen/day (g)
112.53
113.07
112.66
113.35
112.59
0.12
0.220
% HDP*
58.25b
58.47b
62.92a
57.50b
62.04a
0.98
0.040
FCR*
2.34
2.33
2.15
2.37
2.18
0.04
0.350
a,b,c – Means on the same row having different superscripts are significantly (p < 0.05) different.
*SEM = standard error of the mean; FI = feed intake; %HDP = percentage hen day production; FCR =
feed conversion ratio.
Haematological parameters
The haematological profiles of laying hens were not significantly affected by the inclusion of
phytogenic feed additives, with no differences observed across treatments (p > 0.05). Despite
the lack of statistical significance, some numerical variations were noted. Birds fed Diet 2
exhibited higher erythrocyte sedimentation rate (ESR) and lymphocyte (LYM) values compared
to the other groups, while the lowest ESR and LYM were recorded in birds receiving Diets 4
and 1, respectively. Diet 4 resulted in the highest values of packed cell volume (PCV), red blood
cells (RBC), haemoglobin (Hb), and eosinophils (EOS), whereas Diet 2 showed the lowest values
for PCV, RBC, and Hb. The Monocyte (MON) percentage was slightly higher in Diet 1 and lowest
in Diet 4. Detailed values are presented in Table 4.
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Table 4. Effect of phytogenic feed additives on haematology of laying hens.
Parameters
Diet 1
Diet 2
Diet 3
Diet 4
Diet 5
±SEM*
P value
ESR* (mm/h)
3.78
4.33
3.78
3.67
3.89
0.12
0.410
PCV* (%)
26.11
25.00
26.00
26.22
25.78
0.25
0.550
RBC* (× 106 mm3)
2.12
1.87
2.09
2.19
2.06
0.07
0.620
Hb* (g/100mL)
8.68
8.30
8.62
8.71
8.54
0.08
0.540
LYM* (%)
59.11
60.22
59.56
59.56
59.89
0.16
0.270
MON* (%)
12.56
12.33
12.00
11.56
11.67
0.28
0.770
EOS* (%)
1.33
1.22
1.11
1.56
1.33
0.11
0.790
*SEM = standard error of the mean; ESR = erythrocyte sedimentation rate; PCV = packed cell volume;
RBC = red blood cell; Hb = haemoglobin; LYM = lymphocyte; MON = monocytes; EOS = eosinophil.
Serum biochemical indices
No significant differences (p > 0.05) were observed in the serum biochemical parameters of
aged laying hens fed diets containing phytogenic feed additives. However, some numerical
trends emerged across treatments. Birds receiving Diet 1 exhibited higher values of alkaline
phosphatase (ALP), total protein, and globulin compared to those on the supplemented diets.
The highest aspartate aminotransferase (AST) and cholesterol concentrations were recorded
in birds fed Diet 5, whereas the lowest values for both parameters were observed in birds fed
Diet 3. Complete results are presented in Table 5.
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Table 5. Effect of phytogenic feed additives on blood serum parameters.
Parameters
Diet 1
Diet 2
Diet 3
Diet 4
Diet 5
±SEM*
P value
AST* (IU/L)
48.50
39.75
36.00
47.00
70.75
7.70
0.660
ALT* (IU/L)
20.75
60.75
29.25
28.25
10.25
5.75
0.120
PO3* (IU/L)
284.65
151.28
224.58
204.00
254.43
16.34
0.160
Protein (g/L)
50.80
47.08
47.08
39.50
44.55
3.28
0.860
Albumin (g/L)
18.75
27.00
20.50
20.00
21.00
1.16
0.240
Globulin (g/L)
32.05
17.58
26.58
19.60
23.05
2.66
0.470
Urease (mmol/L)
1.75
3.28
2.33
2.33
3.63
0.25
0.150
Creatinine (umol/L)
55.00
74.50
59.50
69.75
77.50
3.93
0.350
Cholesterol (mmol/L)
2.70
2.73
2.65
3.08
3.43
0.18
0.610
HDL* (mmol/L)
2.28
2.73
1.95
2.85
2.88
0.15
0.240
LDL* (mmol/L)
2.73
2.78
2.43
3.10
3.28
0.21
0.720
Triglycerides (mmol/L)
6.05
7.85
4.63
7.08
7.30
0.55
0.400
*SEM = standard error of the mean; AST = Aspartate aminotransferase; ALT = alanine aminotransferase;
PO3 = alkaline phosphate (ALP); HDL = high density lipoprotein; LDL = low density lipoprotein.
Antioxidant properties
Dietary inclusion of phytogenic feed additives significantly (p < 0.05) influenced the antioxidant
parameters in aged laying hens. Clear differences were observed across treatments. Diet 2
exhibited the highest glutathione (GSH) concentrations, while Diet 1 showed the lowest.
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Catalase activity peaked in hens receiving Diet 5 and was lowest in those fed Diet 3. Similarly,
superoxide dismutase (SOD) activity was substantially higher in birds fed Diet 5, while birds on
Diets 1, 3, and 4 exhibited comparably lower values. Detailed results are presented in Table 6.
Table 6. Antioxidant effect of phytogenic feed additives of laying hens.
Parameters
Diet 1
Diet 2
Diet 3
Diet 4
Diet 5
±SEM*
P value
GSH* (uM/mL)
0.21d
2.02a
1.49b
0.53c
0.21d
0.03
0.001
Catalase (uM/mg protein)
0.32c
0.32c
0.19d
0.33b
0.80a
0.01
0.001
SOD* (%)
10.00b
20.00b
10.00b
10.00b
60.00a
2.31
0.001
a,b,c – Means on the same row having different superscripts are significantly (p < 0.001) different. *SEM
= standard error of the mean, GSH = glutathione peroxidase, SOD = superoxide dismutase.
DISCUSSION
In evaluating the nutritional properties of the experimental ingredients, the proximate analysis
of the phytogenic feed additives (PFAs) highlights the natural compositional variability inherent
to plant-based ingredients, driven by factors such as maturity stage, environment, and
processing methods (Ikpeama et al., 2014; Ahaotu and Lawal, 2019). While not intended as
primary protein sources, their crude protein content suggests a supplementary role in
monogastric diets, contributing to overall amino acid availability and dietary balance
(Ogbuewu et al., 2014).
Regarding mineral composition, ash content —an important factor influencing bone
mineralization and eggshell strength— ranged from 3.26% to 6.11% in the tested PFAs,
indicating a meaningful mineral contribution (Javadi et al., 2021). Notably, Diet 4, with the
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highest ash value, may enhance dietary mineral availability. This finding is consistent with Kostić
et al. (2023), who reported that the combination of turmeric and garlic yields synergistic effects
on mineral composition.
In the context of dietary fiber, crude fiber (CF) is essential in poultry nutrition for maintaining
gut health and promoting the digestion and absorption of nutrients (Jiménez-Moreno et al.,
2016). According to NRC (2012), feedstuffs with CF levels above 18% are considered roughages
and are poorly digested by monogastric animals. The CF levels observed in the PFAs studied
(10.28–13.79%) fall within a tolerable range, supporting their use as dietary supplements
without impairing nutrient digestibility in laying hens.
The ether extract values, particularly the highest level in Diet 2 (14.98%), are consistent with
previous reports on turmeric's rich content of beta-carotene, polyphenols, essential fatty acids,
and volatile oils (Ikpeama et al., 2014). These bioactive compounds contribute to the elevated
energy value of PFA-based diets. As noted by Babayemi and Bamikole (2006), such
phytochemicals can enhance energy availability and overall efficiency in poultry systems.
Nitrogen-free extract (NFE), representing the carbohydrate portion of feed, serves as a vital
energy source for poultry (Moniruzzaman and Fatema, 2022; Ndelekwute et al., 2019). The NFE
values reported support the notion that PFAs contribute to growth performance, body
composition, and glucose metabolism (Ge et al., 2023). Furthermore, the presence of plant-
derived bioactive compounds may enhance metabolic pathways and improve energy
utilization, thereby helping maintain overall nutritional balance (Murugesan et al., 2015).
Although no significant effects were observed on performance parameters and serum
biochemical indices, notable improvements were seen in hen-day production (HDP) and
antioxidant enzyme activities. These findings indicate that PFAs contributed to improve
production efficiency and oxidative balance. While feed intake (FI) and feed conversion ratio
(FCR) remained unaffected, HDP was significantly higher in birds receiving Diets 3 and 5. This
enhancement may be linked to improved nutrient utilization and metabolic activity, potentially
mediated by compounds such as gingerols and shogaols, which are known to enhance
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gastrointestinal function and enzymatic activity, thereby facilitating more efficient nutrient
absorption (El-Kashef, 2022; Soliman et al., 2021; Olaniyi et al., 2020; Ayeni et al., 2020;
Banerjee, 2018; Oleforuh-Okoleh et al., 2014).
Haematological indices remained stable across all treatment groups, indicating that PFA
supplementation had no adverse effects on blood health or physiological status in laying hens.
This finding is consistent with previous studies showing that turmeric and ginger
supplementation does not significantly alter hemoglobin concentration, erythrocyte
sedimentation rate, or packed cell volume (Bounous and Stedman, 2000; Dacie and Lewis, 1991;
Hosseini-Vashan et al., 2012; Basavaraj et al., 2011; Sugiharto et al., 2011).
Similarly, serum biochemical parameters—including aspartate aminotransferase (AST), alanine
aminotransferase (ALT), alkaline phosphatase (ALP), total protein, albumin, and cholesterol—
remained within physiological reference ranges. Specifically, the ranges for AST (36.00–70.75
IU/L), ALT (10.25–60.75 IU/L), ALP (151.28–284.65 IU/L), total protein (39.50–50.80 g/L), albumin
(18.75–27.00 g/L), and cholesterol (2.65–3.43 mmol/L) were all within or near the normal limits
for laying hens, as reported by Kaiser et al. (2022) and Kraus et al. (2021).
Although ALT in Diet 2 (60.75 IU/L) slightly exceeded the typical upper limit, previous research
has documented ALT values up to 60 IU/L in healthy birds (Lumeij, 2008). Furthermore, in avian
species, ALT is not the primary indicator of liver function, whereas AST and ALP are more
reliable markers (Kaneko et al., 2008). Given that both AST and ALP remained within normal
ranges and that no clinical signs of hepatic dysfunction were observed, the elevated ALT level
does not suggest liver impairment. Additionally, the trend of reduced ALP values in the
treatment groups may indicate hepatoprotective effects, though further investigations,
including metabolomic profiling, histological evaluation, and longitudinal studies, are needed
to confirm this hypothesis (Onu, 2010).
Antioxidants are essential for protecting against oxidative stress, cellular damage, and chronic
degenerative diseases (Sreelatha and Padma, 2009). A key outcome of this study was the
significant increase in antioxidant enzyme activities, particularly glutathione peroxidase (GSH-
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Px) and catalase, in hens receiving PFA-enriched diets. The highest levels of these enzymes
were observed in birds fed Diets 2 and 5, suggesting that the bioactive compounds in these
additives strengthen the birds’ antioxidant defense mechanisms.
Turmeric, for instance, is rich in curcuminoids (especially curcumin), which possess potent
antioxidant and anti-inflammatory properties through free radical scavenging and modulation
of inflammatory pathways (Rajesh and Devvrat, 2018). Ginger contains gingerols and shogaols,
which enhance gastrointestinal health and stimulate antioxidant enzyme activity, thereby
improving nutrient absorption and metabolic performance (El-Kashef, 2022; Banerjee, 2018;
Morakinyo et al., 2011). Garlic provides allicin and other organosulfur compounds that support
lipid metabolism and reinforce antioxidant responses, thus mitigating oxidative stress
(Navidshad et al., 2018; Sallam et al., 2004).
The synergistic interaction of these phytochemicals likely explains the observed improvements
in antioxidant status and overall physiological condition of the hens. Consistent with these
findings, Hosseini-Vashan et al. (2012) also reported enhanced GSH-Px activity in chicks fed
turmeric-supplemented diets. Therefore, PFA supplementation appears promising for
boosting glutathione levels and maintaining a robust antioxidant system, which is essential for
sustaining health and optimizing productivity in laying hens (Mohammed et al., 2022).
FINAL CONSIDERATIONS
The findings of this study indicate that the inclusion of phytogenic feed additives (PFAs),
notably ginger, turmeric, and garlic, can positively influence both productive performance and
oxidative balance in 64-week-old laying hens. Diets supplemented with ginger, either alone or
in combination with garlic, significantly enhanced hen-day production (HDP) without
negatively affecting feed intake or feed conversion ratio, suggesting improved nutrient
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utilization and metabolic efficiency. These outcomes support the practical application of
ginger-based additives in commercial egg production systems, particularly in aged layers
where performance tends to decline.
The effects in antioxidant enzyme activities, especially glutathione peroxidase and catalase,
detected in hens fed diets containing turmeric or the ginger–garlic blend, are likely attributable
to bioactive phytochemicals such as gingerols, curcuminoids, and allicin, known for their
capacity to modulate oxidative stress responses. The observed enhancement in antioxidant
defense highlights the potential of these additives to improve physiological resilience under
oxidative or metabolic challenges.
Importantly, haematological and serum biochemical profiles indicated the absence of adverse
effects and supporting the safety of these PFAs in layer nutrition. Additionally, subtle trends
suggesting improved liver function in supplemented groups warrant further investigation to
clarify underlying mechanisms. Overall, the use of ginger, alone or synergistically with garlic,
emerges as a promising strategy to improve both performance and health in laying hens.
Future research should focus on elucidating the molecular pathways through which these
compounds exert their effects and refine optimal inclusion rates for application in various
production settings.
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