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Comparative Evaluation of Selected Medicinal Plant Extracts on
Health Biomarkers and Antimicrobial Activity in Broiler Chickens

Halimatu Sadiya Abdullahi*, Moses Zira Zaruwa, Ijeomah Ann Ukamaka, Chibuzo Carole Nweze

Department of Biochemistry. Nasarawa State University, Keffi. Nasarawa State, Nigeria.

Corresponding author*

DOI: https://doi.org/10.51583/IJLTEMAS.2025.1410000014

Received: 28 Sep. 2025; Accepted: 06 Oct. 2025; Published: 30 October 2025

Abstract: This study presents a comparative evaluation of the phytochemical composition and health-promoting potential of seven
medicinal plant extracts Azadirachta indica, Curcuma longa, Allium sativum, Ocimum gratissimum, Vernonia amygdalina, Aloe
barbadensis, and Moringa oleifera on the health biomarkers of broiler chickens. By integrating phytochemical profiling,
biochemical assessment, hematological indices, and antimicrobial testing, this research provides a multi-dimensional understanding
of phytogenic addictive. Phytochemical screening confirmed the presence of phenols, tannins, alkaloids, flavonoids, saponins, and
cardiac glycosides in varying concentrations. Among the extracts, Ocimum gratissimum and Azadirachta indica contained the
highest flavonoid and saponin levels, respectively. Vernonia amygdalina and Moringa oleifera exhibited superior effects on liver
and kidney biomarkers, enhancing total protein while reducing urea and creatinine levels. Allium sativum notably improved
hematological parameters such as hemoglobin concentration, while Azadirachta indica increased total white blood cell counts.
Antimicrobial screening demonstrated that Moringa oleifera and Vernonia amygdalina possessed the strongest inhibition against
pathogenic bacteria, including Salmonella spp., E. coli, and Klebsiella spp. These findings reveal the synergistic potential of
medicinal plants as phytogenic feed additives to improve organ health, immunity, and microbial safety in poultry production. The
comparative approach enhances understanding of plant specific effects and supports development of natural alternative to antibiotics
in broiler chicken.

Key words: Phytochemicals, Broiler Chickens, Liver and Kidney function, Hematological Indices, Antimicrobial Activity,
Antibiotic alternative.

I. Introduction

Medicinal plants remain the most abundant bioresources used for therapeutic purposes in both human and veterinary medicine.
Their increasing application in livestock production, particularly as phytogenic feed additives, addresses the urgent need to reduce
dependence on synthetic antibiotics that contribute to microbial resistance and residues in animal products. Numerous studies have
explored individual plant species such as Moringa oleifera and Azadirachta indica for their growth-promoting and
immunomodulatory properties.. However, comparative assessments examining multiple plants simultaneously remain scarce. This
study aims to fill that gap by conducting a comparative evaluation of seven medicinal plants commonly utilized in traditional
African ethnoveterinary practice. By analyzing their phytochemical composition, biochemical impacts, hematological responses,
and antimicrobial efficacy, this research provides integrated evidence for their collective and differential contributions to broiler
chicken health.

II. Materials and Methods

Ethical Clearance: Ethical approval was obtained from the Ethical Committee, Nasarawa State University, Keffi.

Preparation of Plant Extract

Azadirachta indica, Curcuma longa, Allium sativum, Ocimum gratissimum, Vernonia amygdalina, Aloe barbadensis, Moringa
oleifera fresh leaves were obtained from local farmers in Keffi, area of Nasarawa state, Nigeria. The whole plants were washed
under running tap water, shade dried at room temperature, and powdered. The powdered plant samples (500g/150ml) of solvent
was added and kept for 3 days. The extract was filtered using Whatman No.1 filter paper and the supernatant was collected. The
residue was again extracted two times (with 3 days of interval for each extraction) and supernatants were collected. The supernatants
were evaporated in water bath 28 ± 1°C until the volume was reduced to 150 ml. Extract of the whole plant powder was prepared
and stored in air tight bottles for subsequent analysis.

Phytochemical Analysis

The phytochemical analysis were carried out on the plant extracts using the standard procedures. The presence or absence of a
particular phytochemical compound involved the addition of appropriate standard chemicals/reagents in appropriate sequence to
the plant extract. The following classes of phytochemicals were screened for: phenols, tannins, alkaloids, flavonoids, cardiac
glycosides, saponins, terpenoids, steroids, glycosides

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Uv- Vis Spectrum Analysis

The extract was centrifuged at 3000rpm for 10min and filtered through Whatmann No.1 filter paper. The sample was diluted to
1:10 with the same solvent. The extract was scanned at wave length ranging from 200 to 1100 nm using Perkin Elmer
Spectrophotometer and the characteristic peaks were detected. The peak values of the UV-VIS were recorded.

Experimental Design

A total of 32 healthy day old broiler chicks were purchased from a reputable veterinary shop in Keffi, Nasarawa State and housed
at the animal house of the Department of Biochemistry, Nasarawa State University, Keffi. The birds were acclamatized for two
week before grouping and two weeks after the grouping the treatment commenced and lasts for another two weeks. The broiler
chicken were grouped as follows;

Group 1 (control): Received 2L of water + 50g of normal feed

Group 2: Received of 300mg/kg bw Vernonia amygdalina extract in 2L of water + 50g of normal feed

Group 3: Received 200mg/kg bw of Azadirachta indica extract in 2L of water + 50g of normal feed.

Group 4: Received 330mg/kg of Ocimum gratissimum extract in 2L of water + 50g of normal feed.

Group 5: Received 400mg/kg of Allium sativum extract in 2L of water + 50g of normal feed

Group 6: Received 220mg/kg bw of Curcuma longa extract in 2L of water + 50g of normal feed.

Group 7: Received 500mg/kg bw of Aloe barbadensis extract in 2L of water + 50g of normal feed.

Group 8: Received 500mg/kg bw of Moringa oleifera extract in 2L of water + 50g of normal feed.

Blood Sample Collection

The animals were sacrificed in accordance with the guidelines of the European Convention for the Protection of Vertebrate Animals
and other Scientific Purposes ETS-123 (European Treaty Series, 2005). Whole blood was collected using sterile syringe and needle.
The blood sample of each chicken was collected into respective capped tubes and was centrifuged at 1000 rpm for 10 minutes using
a bench top centrifuge to separate cells from serum and stored in the refrigerator for further analysis.

Biochemical Analysis

The serum obtained was used to analyse for Liver function parameter (Alanine transaminase (ALT), Aspartate transaminase (AST),
Alkaline phosphatase (ALP), Protein following standard mehods of Reitman and Frankel (1957) and kidney function parameters
(Sodium ion (Na+), Potassium ion (K+), Urea and Creatinine) following standard methods of Bartels and Bohmer (1972) with
modification as outlined in Randox kit.

Haematologica Analysis

Whole blood samples were used for haemological parameters (white Blood cell (WBC), Red blood cell (RBC), Pack cell volume
(PCV), Hemoblobin (Hb), mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular
hemoglobin concentration (MCHC) and white cell differentials; Neutrophil (N), Leukocyte (L), Mesophil (M), Esnophil (E))and
Basophil (B) Following standard methods of Williams et al., (1972).

Microbiological Analysis

Plant extracts were inoculated in agar plates, allowing the bioactive components to diffuse into the agar and interact with the
microorganisms. The growth of the microorganisms is inhibited by the various plant extract creating a clear zone around the disk.
The size of the zone of inhibition is measured, indicating the bioactive components in the sample.

Statistical Analysis

The data obtained were analyzed using one-way ANOVA in IBM SPSS version 23.0 to get the means and standard deviations
(n=4). Further test for levels of significance was considered statistically at a level of p<0.05 (Duncan, 1957).

III. Results

Table 1: Qualititative phytochemical screening of Different Plant Extracts

Metabolites C. longa V. amygdalina M.
oleifera

O. gratissimum A. indica A. barbadensis A. sativum

Phenols + + + + ++ + +

Tannins + + + + + + +

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Alkaloids +++ + + + + + +

Flavonoids ++ + + + +++ + +

Cardiac
Glycosides

+ + + + ++ - +

Saponnins ++ + + + +++ + +

Terpenoids - + + + +++ - +

Steroids ++ + + - + - -

Glycosides - - + - - + +

Key: + = Present; ++ = Moderate; +++ = Abundant; – = Absent.
Highest phytochemical abundance observed in A. indica (saponins, terpenoids) and C. longa (alkaloids).

Table 2. Quantitative Phytochemical Concentrations (g/mg) of Different Plant Extracts

Metabolite C. longa V.
amygdalina

M. oleifera O.
gratissimum

A. indica A.
barbadensis

A. sativum

Phenols 8.60 ± 0.51 253.56 ± 2.37 171.33 ±
3.06

531.44 ± 4.57 43.79 ± 1.72 34.94 ± 0.52 39.10 ±
0.45

Tannins 11.29 ±
0.92

76.89 ± 1.44 47.18 ± 1.54 175.43 ± 4.49 10.52 ± 1.11 25.49 ± 1.43 24.24 ±
0.54

Alkaloids 75.44 ±
0.52

76.86 ± 2.68 98.69 ± 0.42 86.88 ± 2.06 28.84 ± 0.43 32.82 ± 1.07 29.74 ±
0.73

Flavonoids 40.74 ±
0.63

232.85 ± 2.05 30.67 ± 2.52 298.03 ± 2.82 199.46 ±
1.01

17.97 ± 0.84 73.64 ±
1.10

Saponins 47.19 ±
2.00

282.96 ± 2.34 82.33 ± 1.25 161.54 ± 2.26 677.95 ±
4.84

11.68 ± 1.41 21.09 ±
0.70

Terpenoids – 21.03 ± 1.49 40.89 ± 0.77 23.47 ± 1.27 161.75 ±
1.54

– 15.80 ±
0.69

Highlight: O. gratissimum exhibited the highest total phenolic and flavonoid content, while A. indica contained the highest saponin
and terpenoid concentrations.

Table 3. Liver Function Parameters in Broiler Chickens Administered with Different Plant Extracts

Group Total Protein (g/L) ALP (UI/L) ALT (UI/L) AST (UI/L) Interpretation

Control 56.00 ± 10.47 310.50 ± 27.99 15.60 ± 6.08 169.30 ± 17.35 –

V. amygdalina 75.38 ± 11.90 269.38 ± 29.89 12.78 ± 0.47 150.54 ± 8.56 ↑ protein, hepatoprotective

A. indica 63.18 ± 4.65 220.00 ± 6.16 13.95 ± 2.57 155.34 ± 12.17 ↓ ALP, hepatoprotective

O. gratissimum 69.50 ± 8.57 84.30 ± 3.35 12.36 ± 5.22 131.10 ± 0.82 Strong hepatoprotection

A. sativum 79.68 ± 6.55 315.75 ± 4.35 11.83 ± 3.54 149.08 ± 3.72 ↑ protein synthesis

C. longa 55.38 ± 8.99 250.40 ± 37.76 12.33 ± 2.05 160.83 ± 1.72 Mild effect

A. barbadensis 58.65 ± 6.60 84.50 ± 4.50 16.38 ± 1.64 133.42 ± 0.47 Mild improvement

M. oleifera 59.38 ± 8.44 237.70 ± 15.93 16.58 ± 4.64 171.03 ± 6.16 Moderate effect

Best performers: V. amygdalina, A. indica, and O. gratissimum showed strongest hepatoprotective effects.

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Table 4. Kidney Function Parameters in broiler chickens Administered with Different Plant Extracts

Group Na⁺
(mmol/L)

K⁺
(mmol/L)

Urea
(mmol/L)

Creatinine
(mmol/L)

Interpretation

Control 167.20 ±
3.11

3.75 ± 0.35 7.30 ± 0.57 69.40 ± 5.51 –

V. amygdalina 139.05 ±
0.82

3.59 ± 0.09 3.11 ± 0.09 42.68 ± 5.19 Strong
nephroprotection

A. indica 137.75 ±
1.21

3.88 ± 0.22 3.33 ± 0.25 59.68 ± 1.81 Improved kidney
function

M. oleifera 166.28 ±
2.58

3.63 ± 0.53 2.20 ± 0.08 49.83 ± 4.26 Best renal
improvement

C. longa, O. gratissimum, A.
sativum, A. barbadensis

– – – – Mild-to-moderate
effects

Key: Best performers-M. oleifera > V. amygdalina > A. indica for renal protection.

Table 5. Hematological Parameters of Broiler Chickens Administered with Different Plant Extracts

Parameter Control V.
amygdalina

A. indica A.
sativum

M.
oleifera

Key Observation

WBC (×10⁹/L) 4.10 ±
0.42

6.56 ± 1.41 26.63 ±
9.35

6.58 ±
0.57

13.13 ±
6.46

A. indica markedly increased WBC,
indicating enhanced immune response

Hb (g/dl) 11.55 ±
0.49

10.99 ± 2.33 14.68 ±
1.25

16.93 ±
0.97

13.93 ±
0.31

A. sativum showed highest hemoglobin
value, enhancing oxygen transport

RBC (×10¹²/L) 4.40 ±
0.14

4.73 ± 0.13 5.58 ±
0.62

6.35 ±
0.21

5.43 ±
0.46

A. sativum stimulated erythropoiesis

PCV (%) 37.00 ±
1.41

33.75 ± 6.24 47.25 ±
1.71

31.25 ±
2.22

42.25 ±
1.71

A. indica improved packed cell volume

MCV (fL) 83.50 ±
0.71

124.00 ±
10.13

233.75 ±
15.19

114.73 ±
5.59

69.00 ±
2.94

A. indica improved erythrocyte
morphology

MCH (pg) 26.45 ±
0.21

41.14 ± 3.68 43.50 ±
1.29

36.63 ±
1.60

22.75 ±
0.96

A. indica and A. sativum enhanced
hemoglobin content

MCHC (g/dl) 31.35 ±
0.07

33.26 ± 0.05 33.00 ±
0.61

33.20 ±
0.00

33.00 ±
0.82

Stable across groups, slight
improvement noted

Neutrophils
(%)

5.00 ±
1.41

11.93 ± 3.84 33.25 ±
2.22

32.25 ±
3.40

11.25 ±
1.71

A. indica induced strong immune
activation

Lymphocytes
(%)

85.00 ±
2.83

67.25 ± 1.71 60.75 ±
1.71

62.00 ±
1.63

81.75 ±
2.22

Lymphocyte balance maintained

Eosinophils
(%)

2.50 ±
0.71

2.75 ± 0.96 1.50 ±
0.58

2.75 ±
0.50

2.50 ±
0.58

No significant variation

Interpretation: Allium sativum enhanced erythropoietic parameters (Hb, RBC), while A. indica improved immune and hematological
indices (WBC, neutrophils, PCV).

Table 6. Antimicrobial Activity of Plant Extracts (Zone of Inhibition)

Bacterial Isolate V.
amygdalina

A.
indica

O.
gratissimum

C.
longa

M.
oleifera

Interpretation

Salmonella spp. 26mm 26mm 25mm 24mm 25mm Broad-spectrum inhibition observed
across extracts

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E. coli 24mm 26mm 27mm 26mm 26mm High activity against Gram-negative
bacteria

Klebsiella spp. 24mm 0mm 24mm 26mm 27mm Strongest inhibition by M. oleifera

Pseudomonas spp. 27mm 26mm 26mm 24mm 22mm Effective control by V. amygdalina
and A. indica

Staphylococcus
aureus

27mm 26mm 0mm 25mm 27mm Potent Gram-positive inhibition by M.
oleifera and V. amygdalina

Key: Moringa oleifera and Vernonia amygdalina demonstrated the broadest antimicrobial spectra, effectively inhibiting both Gram-
positive and Gram-negative bacteria, followed by A. indica.

V. Discussion

The comparative analysis of the seven plant extracts revealed that each species contributed differently to the physiological and
biochemical well-being of broiler chickens. The high levels of flavonoids and phenolic compounds (Table1) observed in Ocimum
gratissimum and Azadirachta indica corroborate findings from Zeng et al. (2015) and Perez-Gregorio et al. (2021), who reported
that flavonoid-rich phytobiotics enhance liver detoxification and oxidative stability in poultry.

Vernonia amygdalina and Moringa oleifera (Table 3 and 4) demonstrated marked improvements in liver and kidney function,
consistent with studies from Agbetuyi and Oloruntola (2020) and Mohammed et al. (2021), suggesting their hepatoprotective and
nephroprotective properties.

Vernonia amygdalina and Moringa oleifera demonstrated marked improvements in liver and kidney function, consistent with
studies from Agbetuyi and Oloruntola (2020) and Mohammed et al. (2021), suggesting their hepatoprotective and nephroprotective
properties. The increase in serum protein levels indicates improved protein metabolism and nutrient utilization, critical for growth
performance and immune competence.

The hematological results (Table 5) also align with previous reports that Allium sativum enhances erythropoiesis and immune cell
activation due to its organosulfur compounds. Elevated WBC counts following Azadirachta indica supplementation suggest
enhanced immune readiness, consistent with global reports on neem’s immunomodulatory capacity (Khater et al., 2020).

The broad-spectrum antimicrobial activity (Table 8), particularly from Moringa oleifera and Vernonia amygdalina, supports their
traditional use as natural antimicrobials. Similar inhibition patterns have been observed in Asian and European poultry trials,
reinforcing the global potential of these plants as safe antibiotic alternatives. This study therefore provides novel comparative
insights, demonstrating that the simultaneous assessment of multiple phytogenic plants offers a more holistic understanding of
functional diversity.

VI. Conclusions and Future Perspectives

The findings of this study confirm that medicinal plants such as Moringa oleifera, Vernonia amygdalina, Azadirachta indica, and
Allium sativum provide complementary health benefits when used as phytogenic feed additives in broiler chickens. They enhance
liver and kidney function, improve hematological profiles, and exhibit potent antimicrobial activity. The comparative approach
strengthens the understanding of their differential effects and highlights their collective potential to promote poultry production.

Future studies should focus on larger sample sizes, longer trial durations, and inclusion of growth performance, carcass quality, and
economic analyses to establish commercial feasibility. Integrating metabolomic and genomic tools could further elucidate the
molecular mechanisms underlying these beneficial effect.

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