Archives of Immunology Research and Therapy

Abstract

A 42-day feeding trial was conducted to evaluate the ameliorative potential of Afrocarpus falcatus leaf powder on the growth performance and hematological indices of broiler chicks exposed to Aflatoxin B1 (AFB1). A total of 200 one day-old broiler chicks were randomly assigned to 4 dietary treatments in a Completely Randomized Design (CRD). Each treatment was replicated 5 times with 10 birds per replicate, housed in a sanitized battery cage system. Treatment 1 (T1) served as the absolute control, receiving a standard basal diet without toxin or additives. Treatment 2 (T2) was fed a basal diet contaminated with 0.5 g AFB1/ kg feed and supplemented with 250 mg synthetic ascorbic acid/kg. Treatments 3 (T3) and 4 (T4) were fed the 0.5 g AFB1 /kg contaminated diet supplemented with Afrocarpus falcatus leaf powder at 150 g/kg and 200 g/kg of feed, respectively. At the end of the 42-day experimental period, final body weight (FBW) was significantly lower (P<0.05) in T2 compared to T1, T3, and T4, which maintained statistically similar weights. Feed intake (FI) followed a distinct trend, being highest in T1, intermediate in T3 and T4, and lowest in T2 (P<0.05). The feed conversion ratio (FCR) was significantly higher (P<0.05) in the unprotected T2 group, whereas T3 and T4 effectively restored FCR values to par with the T1 control. Hematological analysis revealed that Red Blood Cell (RBC) counts, Packed Cell Volume (PCV), and Hemoglobin (Hb) concentrations remained within the normal physiological reference range for broilers across all groups; however, values were significantly lower (P<0.05) in the T2 group than in T1, T3, and T4. Total White Blood Cell (WBC) counts were not significantly altered (P>0.05) by any of the dietary interventions. In conclusion, dietary exposure to 0.5 g AFB1/kg causes severe growth depression and subclinical microcytic anemia in broilers, which cannot be fully mitigated by 250 mg of ascorbic acid. However, dietary inclusion of Afrocarpus falcatus leaf powder at both 150 g/kg and 200 g/kg effectively neutralizes AFB1 toxicity, protecting erythrocytic indices and restoring growth performance parameters to control levels. Afrocarpus falcatus shows strong potential as a functional phytogenic feed additive for organic mycotoxin management in commercial poultry production.

Introduction

The global poultry industry plays a critical role in meeting human demand for high-quality animal protein, with broiler production serving as one of its most dynamic and fastgrowing sectors [1]. To keep pace with this demand, commercial producers rely heavily on intensive feeding practices and standardized management protocols [2]. However, the economic sustainability of intensive broiler production is frequently threatened by the contamination of feed ingredients with mycotoxins [3-4]. Among these, Aflatoxin B 1 (AFB1) a highly toxic secondary metabolite primarily synthesized by the ubiquitous fungal species Aspergillus flavus and Aspergillus parasiticus is considered the most prevalent and economically devastating threat in tropical and subtropical regions [4]. In commercial broiler facilities, the co-exposure of intensive production systems and compromised feed hygiene creates a high-risk environment for systemic mycotoxicity [5-6]. When broilers ingest AFB1-contaminated rations, it triggers severe economic and biological failures, commonly characterized by structural tissue damage, reduced growth, and suppressed physiological function [7]. Intensive broiler production systems require optimal health and defense against feed-borne toxins [7].

The core problem facing the poultry sector is that conventional postharvest drying and storage techniques often fail to prevent the proliferation of Aspergillus species in key feed ingredients like corn and soybean meal, especially under hot and humid tropical climates [8]. Once synthesized, AFB1 is exceptionally heatstable and resistant to typical feed manufacturing processes, making its elimination from finished rations a major technical challenge [9]. When consumed by broiler chicks, AFB1 undergo rapid hepatic biotransformation cytochrome P450 enzymes [10]. This pathway generates highly reactive intermediate metabolites, most notably aflatoxin-B1 -8, 9-epoxide. This electrophilic intermediate binds covalently to cellular DNA, forming toxic adducts that disrupt protein translation and transcription [10]. At the systemic level, this biochemical disruption manifests as severe hepatotoxicity, lipid peroxidation of intestinal and erythrocytic biomembranes, and suppressed nutrient assimilation [11]. Consequently, affected flocks experience a drop in feed intake, stunted body weight gain, and high feed conversion ratios (FCR) [11]. Furthermore, the induced oxidative stress destabilizes mature red blood cell membranes, leading to accelerated hemolysis, anemia, and altered hematological profiles [12]. This reduces production margins and causes significant economic losses worldwide [13].

To mitigate the destructive impacts of aflatoxicosis, historical research has evaluated various therapeutic interventions, including synthetic chemical binders, inorganic clays, and singlesource vitamins [14]. For example, supplementing diets with synthetic antioxidants like ascorbic acid (Vitamin C) has been a common mitigation strategy due to its role as an electron donor that neutralizes free radicals and supports tissue regeneration [15]. However, previous studies have demonstrated that relying on synthetic ascorbic acid alone offers limited, narrow spectrum protection when animals face a highdose mycotoxin challenge [16]. While synthetic vitamins can partially offset systemic oxidative stress, they fail to provide a physical barrier against toxin absorption in the intestine, nor do they possess the structural complexity needed to fully protect hepatic or hematopoietic tissues from severe AFB1 exposure [15]. As a result, recent livestock research has shifted toward identifying complex, multicomponent phytogenic feed additives [17]. These plant derived alternatives offer broad spectrum protection by combining direct radical scavenging with physical bio sorption properties [17].

Given the limitations of single-molecule synthetic alternatives, there is a clear need to evaluate underexplored, bio-active tropical plants as sustainable, low-cost solutions for managing mycotoxin risks. Afrocarpus falcatus (Outeniqua yellowwood) is a promising candidate for this purpose due to its dense profile of secondary metabolites, including a natural blend of polyphenols, complex tannins, and unique bifla-vonoids such as podocarpusflavone and sciadopitysin [18]. The foliage of Afrocarpus falcatus contains a rich profile of bioactive compounds. Using the whole crude leaf powder of Afrocarpus falcatus provides a dual-action defense against AFB1 toxicity that synthetic alternatives cannot match [18]. The rich concentrations of flavonoids and polyphenols acts as a potent antioxidant network, scavenging reactive oxygen species (ROS) and preventing the lipid peroxidation of erythrocyte and hepatic membranes. The structural, condensed tannins within the leaf powder can form stable complexes with AFB1 within the gastrointestinal tract, reducing its bio-availability and preventing its absorption into the portal circulation [17].

By investigating the inclusion of Afrocarpus falcatus leaf powder at graded levels in comparison to a standard synthetic ascorbic acid treatment, this study provides valuable data on organic mycotoxin management. The findings establish a practical, plant-based strategy to protect the health and performance of broiler chickens, offering a reliable alternative to safeguard poultry production from the economic impact of aflatoxin contamination.

Materials and Methods

Experimental Site, Environmental Conditions, and Ethical Approval

The 42-day feeding trial was executed at the Poultry Section of the Gandhi College of Agriculture, Rajasthan, India. The institutional study site is geographically positioned between latitude 230 03 ′N to 300 12 ′ N and longitude 690 30 ′ E to 78017 ′E. Environmental monitoring throughout the experimental window recorded a mean annual temperature range fluctuating between 25.80 C and 35.10 C, alongside a mean annual rainfall ranging from 1100 mm to 1650 mm. All standard management, handling, and experimental interventions involving live birds were stringently subjected to review and secured formal clearance from the Institutional Animal Care and Ethics Committee under the specific authorization protocol code (AGN/30221/2026189).

Preparation and Processing of Afrocarpus falcatus Leaf Powder

Fresh leaves of Afrocarpus falcatus (Outeniqua yellowwood) were gathered during early morning hours from designated botanical repositories in the region. The harvested leaves were immediately transported to the Botany Department laboratory where their taxonomy was verified and recorded under voucher reference number AF/10002/202F deposit-ed for future verification. The raw leaves were meticulous-ly sorted manually to strip away extraneous organic matter and debris, followed by washing in clean distilled water. The cleaned leaves were evenly distributed across plastic trays and subjected to unheated shade-drying at room temperature for a duration of 12 consecutive days until a constant weight was recorded was achieved. This deliberate thermal preservation strategy prevents the degradation of heat-sen-sitive phytogenic metabolites. The dried leaves were then crushed using a heavy-duty mechanical mill to achieve a homogeneous coarse leaf powder. This mechanical modi-fication maximizes the cellular surface area, optimizing the bioavailability of active polyphenolic matrices, tannins, and complex flavonoids when homogeneously blended into the daily raw feed fractions.

Fungal Inoculation, Extraction, and High-Performance Liquid Chromatography (HPLC) Quantification of Afla-toxin B1

The creation of the target mycotoxin challenge was initiated by using an active, toxigenic strain of Aspergillus flavus ob-tained from the Microbiology Laboratory, Sumitra Research Institute, Gujarat to inoculate a clean batch of cracked white maize matrix. The moisture content of the maize substrate was adjusted to a constant 22 % by adding sterile distilled water, before it was autoclaved at 1210C for 20 minutes to eliminate competing native microbial flora. The sterile substrate was then inoculated with the A. flavus spore suspension and incubated under controlled environmental dark chambers at a constant temperature of 280C and 85% relative humidity for a 14-day production window to stimulate secondary fungal metabolism and optimize the biosynthesis of Aflatoxin B1(AFB1 ).Following the incubation cycle, the fungal activity was halted by steaming the toxic corn matrix at 1000 C for 15 minutes, followed by complete dehydration in a forced-air oven at 600 C until a stable dry mass was achieved. The toxic corn was ground into a uniform meal. The quantification of AFB1 concentrations within the generated substrate meal was determined using High-Performance Liquid Chroma-tography (HPLC) equipped with post-column photochemical derivatization. A 25 g representative sample of the toxic corn meal was extracted using a 100 mL mixture of methanol and water (80:20 v/v) by shaking vigorously on an orbital shaker for 45 minutes.

The crude extract slurry was filtered through Whatman No. 4 filter paper, and a 10 mL aliquot of the filtrate was diluted with 40 mL of phosphate-buffered saline (PBS, pH 7.4). The diluted extract was then passed through an AFB -specific immunoaffinity clean-up column at a flow rate of 1 drop per second to isolate the mycotoxin molecules. The column was washed with 10 mL of pure distilled water, and the bound AFB1molecules were eluted by passing 1.5 mL of HPLC-grade methanol through the matrix.The purified eluate was directly injected into an Agilent 1260 Infinity II HPLC System (Agilent Technologies, United States). The operational configuration utilized a reversed-phase C 18 analytical column (4.6 mm×250 mm, 5 μm particle size) maintained at an internal oven temperature of 350C. The mobile phase consisted of an isocratic blend of water, meth-anol, and acetonitrile (60:20:20 v/v/v) driven at a constant flow rate of 1.0 mL/min. Detection was accomplished using a high-sensitivity fluorescence detector with excitation and emission wavelengths configured precisely at 365 nm and 435 nm, respectively. Using the determined potency of the stock corn meal, precise portions were calculated and incorporated into the basal broiler rations to establish an exact, uniform contamination exposure level of 0.5 g (500 mg) of pure AFB1per kilogram of finished experimental diet across treatments T2, T3, and T4.

Experimental Design, Housing Management

A total of 200 one-day-old broiler chicks (Cobb 500) was purchased from a reputable source in Rajathan, India. Chicks were unboxed and their average weight was recorded using a digital sensitive scale before it was randomly distributed split into 4 distinct dietary treatments, with each individual treatment comprised of 5 independent replicates containing 10 birds each using Completely Randomized Design (CRD). The birds were housed within a strictly sanitized and disinfected multi-tier battery cage system located inside an environmentally regulated poultry house. Prior to the arrival of the chicks, all pre-experimental hygiene procedures were carried out, including thorough pressure washing, chemical disinfection of the floors, cages, and boundaries, and setting up clean feeding and watering troughs. The baseline environmental temperature within the brood-ing zone was initialized at 330 C using automated gas heaters, then gradually scaled down by 2.50 C every week until a steady ambient comfort threshold of 240 C was reached. Clean water and the standard experimental diets which was formulated according to the standard nutritional requirement for broilers [18] was provided on an ad libitum basis through-out the 42-day production cycle. Proximate composition of experimental diet was carried out according to the method outlined by [19].

The experimental feeding groups were formulated as follows:

Treatment 1 (T1): Control group receiving only the standard 

basal diet without any AFB1 contamination or antioxidant additives.

Treatment 2 (T2): Basal diet contaminated with 0.5 g AFB1/ kg diet, supplemented with 250 mg of synthetic ascorbic acid per kilogram of feed.

Treatment 3 (T3): Basal diet contaminated with 0.5 g AFB1/ kg diet, supplemented with 150 g of processed Afrocarpus falcatus leaf powder per kilogram of feed.

Treatment 4 (T4): Basal diet contaminated with 0.5 g AFB1/ kg diet, supplemented with 200 g of processed Afrocarpus falcatus leaf powder per kilogram of feed.

Growth Performance Parameters

Final Body Weight (FBW) and Feed Intake (FI) were mea-sured at the end of the 42-day trial using a high-precision electronic digital platform scale. The Feed Conversion Ratio (FCR) for each replicate was calculated mathematically by dividing the total feed consumed within that replicate by the corresponding total live weight gain recorded over the 42-day timeline.

Blood Collection and Analysis

At the exact termination of the 42-day experiment, 5 birds were randomly chosen from each treatment group for blood profiling. Blood samples (3 mL) were collected via branchial vein puncture using sterile needles and deposited into vacuum tubes containing ethylene diamine tetra acetic acid (EDTA) anticoagulant to preserve cellular integrity. The complete blood count focused on Red Blood Cell (RBC) count, Packed Cell Volume (PCV), Hemoglobin (Hb) concentration, and total White Blood Cell (WBC) count—was performed using a Mindray BC-2800 Vet Automated Hematology Analyzer (Mindray Bio-Medical Electronics Co., Ltd., Shenzhen, China). The analyzer was configured using verified, animal-spe-cific software parameters calibrated for avian blood morphol-ogy to account for the nucleated nature of avian erythrocytes and thrombocytes. The operational specifications of the kit has a sample aspiration volume of 20 μL (whole blood mode); 20 μL (pre-diluted mode), aperture diameter (80 μm) and the measurement principle is via electronic impedance method for cell counting and cyanide-free colorimetric method for hemoglobin quantification.

Statistical Analysis

All data collected on growth performance parameters (Fi-nal Body Weight, Feed Intake, and Feed Conversion Ratio) and hematological indices (RBC, PCV, Hb, and WBC) were subjected to a one-way Analysis of Variance (ANOVA) for a Completely Randomized Design (CRD) using the General

Linear Model (GLM) procedure of SAS (Statistical Analysis System, version 9.4, SAS Institute Inc., Cary, NC, USA).The underlying statistical model used for the analysis is formulated as follows: Y ij =μ+Ti +ϵ ij

Where:

Y ij represents the individual observation of the dependent variable from the j-th replicate within the i-th dietary treat-ment.

μ is the overall population mean for the parameter measured. Ti represents the fixed effect of the i-th dietary treatment (i=1, 2, 3, 4).

ϵij is the random residual error associated with the j-th repli-cate in the i-th treatment, assumed to be independently and identically distributed following a normal distribution: ϵij

Results

Phyto-components in Afrocarpus falcatus leaf powder is presented in Table 2. The plant contained tannins (10.24 mg/g), saponins (35.62 mg/g), phenols (276.33 mg/g), alka-loids (27.11 mg/g), terpenoids (131.4 mg/g), steroids (65.72 mg/g) and flavonoids (290.2 mg/g). In ranking order: phenols > flavonoids > terpenoids > steroids > saponins > alkaloids > tannins.

Growth performance of broiler chickens exposed to Afrocar-pus falcatus leaf powder and a diet contaminated with Afla-toxin B1 (AFB1) is presented in Table 3. Body weight gain was lower in T2 (1763.94 g) than T1 (2406.23 g), T3 (2443.46 g) and T4 (2450.98 g) (p<0.05). Cummulative feed intake was more in T1 compared to the other groups (p<0.05). Feed conversion ratio varied from 2.00 – 2.54, value obtained was higher in T2 relative to the other treatments (p<0.05).

Haematological indices of broiler chickens exposed to Afro-carpus falcatus leaf powder and a diet contaminated with Af-latoxin B1 (AFB1) is presented in Table 4. Whereas pack cell volume, haemoglobin and red blood cell were lower (p<0.05) for T2 than for T1, T3 and T4. White blood cells were similar (p >0.05) among the treatments.

Haematological indices of broiler chickens exposed to Afro-carpus falcatus leaf powder and a diet contaminated with Af-latoxin B1 (AFB1) is presented in Table 4. Whereas pack cell volume, haemoglobin and red blood cell were lower (p<0.05) for T2 than for T1, T3 and T4. White blood cells were similar (p >0.05) among the treatments.

2.5 kg of vitamin-mineral premix contains: Vitamin D3 - 2,000,000 IU; Vitamin K - 2,250 mg; Vitamin A 10,000,000 IU; Vitamin E - 20,000 IU; Thiamine B1 - 1,750 mg; Niacin - 27,500 mg; Pantothen-ic acid - 7,500 mg; Biotin - 50 mg; Choline chloride - 400 g; Ribofla-vin B2 - 5,000 mg; Pyridoxine B6 - 2,750 mg; Antioxidant - 125 g; Magnesium - 80 g; Iodine - 1.2 g; Selenium - 200 mg; Cobalt - 200 mg; Zinc - 50 mg; Iron - 20 g; Copper - 5 g

Table 1: Ingredient and chemical composition of experimental (bas-al) diet

Table 2: Phyto-components in Afrocarpus falcatus leaf powder

Note on Superscripts: ᵃ˒ ᵇ˒ ᶜ Means along the same row with different superscripts are significantly different (p<0.05) Treatment 1 (T1): Control group receiving only the standard basal diet without any AFB1 contamination or antioxidant additives; Treat-ment 2 (T2): Basal diet contaminated with 0.5 g AFB1/kg diet, supplemented with 250 mg of synthetic ascorbic acid per kilogram of feed;

Treatment 3 (T3): Basal diet contaminated with 0.5 g AFB1/kg diet, supplemented with 150 g of processed Afrocarpus falcatus leaf powder per kilogram of feed; Treatment 4 (T4): Basal diet contaminated with 0.5 g AFB1/kg diet, supplemented with 200 g of processed Afrocarpus falcatus leaf powder per kilogram of feed.

Table 3: Growth performance of broiler chickens exposed to Afrocarpus falcatus leaf powder and a diet contaminated with Aflatoxin B1 (AFB1)

Note on Superscripts: ᵃ˒ ᵇ Means along the same row with different superscripts are significantly different (p<0.05) Treatment 1 (T1): Control group receiving only the standard basal diet without any AFB1 contamination or antioxidant additives; Treat-ment 2 (T2): Basal diet contaminated with 0.5 g AFB1/kg diet, supplemented with 250 mg of synthetic ascorbic acid per kilogram of feed; Treatment 3 (T3): Basal diet contaminated with 0.5 g AFB1/kg diet, supplemented with 150 g of processed Afrocarpus falcatus leaf powder per kilogram of feed; Treatment 4 (T4): Basal diet contaminated with 0.5 g AFB1/kg diet, supplemented with 200 g of processed Afrocarpus falcatus leaf powder per kilogram of feed.

Table 4: Haematological indices of broiler chickens exposed to Afrocarpus falcatus leaf powder and a diet contaminated with Aflatoxin B1 (AFB1)

Discussion

The inclusion of Afrocarpus falcatus leaf powder at graded levels (150 g/kg and 200 g/kg) successfully countered the severe systemic damage caused by Aflatoxin B1(AFB1) over the 42-day broiler trial. Ingestion of 0.5 g AFB1/kg without sufficient protection as seen in the T2 group supplemented with only 250 mg of ascorbic acid triggered classic symp-toms of aflatoxicosis [20]. At the cellular level, AFB1 is con-verted by hepatic cytochrome P450 enzymes into the highly reactive intermediate aflatoxin-B1-8, 9-epoxide [21]. This electrophilic compound binds to cellular macromolecules to form toxic DNA adducts, which block transcription and translation [21]. This broad disruption of protein synthesis directly explains the significantly lower final body weights observed in T2 [22]. Furthermore, the unmitigated oxidative stress caused by AFB1generates a surge of reactive oxygen species (ROS) that attack the lipid membranes of the intestinal mucosa [23]. This lipid peroxidation causes the breakdown of intestinal villi, reducing their height and lowering the gut’s overall capacity to absorb nutrients [23-24]. This structural damage, paired with toxin-induced appetite suppression, led to the lower feed intake and elevated feed conversion ratio (FCR) in T2. This demonstrates that the birds consumed feed but could not convert it efficiently into muscle mass because their metabolic resources were redirected toward hepatic detoxification [24]. This result is in agreement with the report of [6] when ginger extract was supplemented in the diet of broiler chicken.

The higher final body weight and feed conversion efficiency in the T3 and T4 groups proves that Afrocarpus falcatus leaf powder provides superior protection against aflatoxicosis compared to synthetic ascorbic acid alone. This impressive therapeutic effect is directly tied to the rich and diverse profile of phytochemicals within the plant’s leaves [2]. The leaf powder contains exceptionally high levels of flavonoids (290.2 mg/g) and phenols (276.3 mg/g). These polyphenolic compounds function as a powerful antioxidants [15-33]. They easily donate hydrogen atoms or electrons to neutralize ROS and lipid peroxyl radicals, halting the oxidative destruction of hepatic and intestinal cell membranes [16]. Additionally, the terpenoids (131.4 mg/g) and steroids (65.72 mg/g) in the leaf powder likely worked alongside these antioxidants to reduce inflammation and stabilize cell walls, protecting the structural integrity of the gut lining [34]. By preserving the intestinal villi, the leaf powder maintained normal nutrient transport and as-similation [35]. This allowed the birds in T3 and T4 to utilize their feed efficiently, resulting in an optimal feed conversion ratio matching the toxin-free control (T1), despite showing an intermediate feed intake. The performance of the leaf powder is further explained by its content of tannins (10.24 mg/g) and alkaloids (27.11 mg/g), which provide an elegant physical defense mechanism within the digestive tract [33]. Tannins are high-molecular-weight polyphenols known for their ability to bind to various chemical complexes [15]. In the small intestine of birds, these tannins can form stable, insoluble hydrophobic complexes with the hydrophobic phenan-threne rings of AFB1. This physical binding effectively traps a portion of the mycotoxin inside the digestive tract, preventing it from crossing the intestinal wall into the portal bloodstream [16].

By acting as a natural bio-sorbent, the leaf powder reduced the total amount of AFB1 absorbed by the birds. This significantly eased the toxic burden on the liver and bone marrow. This dual action where tannins physically block toxin absorption while high concentrations of flavonoids and phenols chemically neutralize systemic oxidative stress explains why Afrocarpus falcatus leaf powder protected the birds far more effectively than the single-molecule, water-soluble ascorbic acid treatment used in T2.

The hematological results provide clear evidence of this systemic protection. The lower Red Blood Cell (RBC) counts, Packed Cell Volume (PCV), and Hemoglobin (Hb) levels in the T2 group indicate a state of sub-clinical microcytic anemia caused by unmitigated aflatoxicosis [25]. This decline happens because AFB1 directly suppresses the bone marrow, slowing down the production of new red blood cells (erythropoiesis) [26]. At the same time, circulating free radicals weaken mature red blood cell walls, causing them to rupture prematurely (hemolysis) [26]. In contrast, the birds in T3 and T4 maintained significantly higher RBC, PCV, and Hb values than those in T2, keeping them well within normal reference ranges [27]. The abundant flavonoids (290.2 mg/g) and saponins (35.62 mg/g) in the leaf powder helped stabilize erythrocyte membranes, protecting them from oxidative damage and hemolysis [27]. Moreover, by protecting liver tissue from damage, the leaf powder ensured the continuous synthesis of key proteins like transferrin, which are essential for normal iron transport and hemoglobin production [28]. Finally, the fact that White Blood Cell (WBC) counts remained statistically unaffected across all groups indicates that the dietary challenge of 0.5 g AFB1/kg did not cause acute immune cell depletion or severe systemic inflammation [29]. This suggests that the primary damage caused by this level of aflatoxin over 42 days is metabolic rather than destructive to white blood cell lines [28]. This result is in agreement with the report of [30].

Conclusion

In conclusion, the outcome of this experiment demonstrate that Afrocarpus falcatus leaf powder contains bioactive compounds with various therapeutic properties. Its inclusion at both 150 g and 200 g per kg of diet successfully ameliorates the destructive effects of Aflatoxin B1 in broiler chickens and protects hematopoietic system from oxidative damage and preserves gut and liver health, allowing for optimal feed conversion, growth performance on par with a toxin-free diet. It represents a highly promising, organic phytogenic feed addi- tive for managing mycotoxin risks in poultry production.

References

1. Alagbe, J. O. “Dietary supplementation of rauvolfia vomitoria root extract as a phytogenic feed additive in growing rabbit diets: growth performance and caecal microbial population.” Concept in Dairy and Veterinary Sciences 4, no. 2 (2021): 2021.
   View at Publisher | View at Google Scholar
2. Sharma, Singh, Alagbe Olujimi John, Liu Xing, Sharma Ram, and Kumar Amita. “Comparative analysis of ethanolic Junipe-rus thurifera leaf, stem bark and root extract using gas chro-matography and mass spectroemetry.” International Journal of Agriculture and Animal Production 2, no. 6 (2022): 18-27.
   View at Publisher | View at Google Scholar
3. Adeyeye S.A, Oloruntola O.D, Ayodele, S.O, Olajide O.O, Abdusalam, L.O, Adeniji, O.E. (2024). Effect of administering aqueous extract of morinda lucida leaves on the performance of broiler chickens fed an aflatoxin b1 contaminated diet. Proc. 49th Conf., Nig. Soc. for Anim. Prod. 24 – 27 March, 2024, Univ. of Ibadan, Nigeria.
   View at Publisher | View at Google Scholar
4. Yunus, Agha W., Ebrahim Razzazi-Fazeli, and Josef Bohm. “Aflatoxin B1 in affecting broiler’s performance, immunity, and gastrointestinal tract: A review of history and contemporary is-sues.” Toxins 3, no. 6 (2011): 566-590.
   View at Publisher | View at Google Scholar
5. Fouad, Ahmed Mohamed, Dong Ruan, HebatAllah Kasem El-Senousey, Wei Chen, Shouqun Jiang, and Chuntian Zheng. “Harmful effects and control strategies of aflatoxin b1 produced by Aspergillus flavus and Aspergillus parasiticus strains on poultry.” Toxins 11, no. 3 (2019): 176.
   View at Publisher | View at Google Scholar
6. Vipin, A. V., Raksha Rao, Nawneet Kumar Kurrey, Anu Appaiah KA, and G. Venkateswaran. “Protective effects of phenolics rich extract of ginger against Aflatoxin B1-induced oxidative stress and hepatotoxicity.” Biomedicine & pharmacotherapy 91 (2017): 415-424.
   View at Publisher | View at Google Scholar
7. Vipin, A. V., Raksha Rao, Nawneet Kumar Kurrey, Anu Appaiah KA, and G. Venkateswaran. “Protective effects of phenolics rich extract of ginger against Aflatoxin B1-induced oxidative stress and hepatotoxicity.” Biomedicine & pharmacotherapy 91 (2017): 415-424.
   View at Publisher | View at Google Scholar
8. Oloruntola, Olugbenga D., Simeon O. Ayodele, Deborah A. Ol-oruntola, Olumuyiwa J. Olarotimi, Andrew B. Falowo, Victor O. Akinduro, Francis A. Gbore, Olufemi A. Adu, and Johnson O. Agbede. “Dietary supplementation of Capsicum powder affects the growth, immunoglobulins, pro-inflammatory cytokines, adi-pokines, meat, and liver histology of aflatoxin B1 exposed broil-er chickens.” Toxicon 240 (2024): 107640.
   View at Publisher | View at Google Scholar
9. Olarotimi, Olumuyiwa Joseph, Francis Ayodeji Gbore, Olufemi Adesanya Adu, Olugbenga David Oloruntola, and Olatunji Abu-bakar Jimoh. “Ameliorative effects of Sida acuta and vitamin C on serum DNA damage, pro-inflammatory and anti-inflamma-tory cytokines in roosters fed aflatoxin B1 contaminated diets.” Toxicon 236 (2023): 107330.
   View at Publisher | View at Google Scholar
10. Jobe, Martha Cebile, Doctor MN Mthiyane, Phiwayinkosi V. Dludla, Sithandiwe E. Mazibuko-Mbeje, Damian C. Onwudiwe, and Mulunda Mwanza. “Pathological role of oxidative stress in aflatoxin-induced toxicity in different experimental models and protective effect of phytochemicals: a review.” Molecules 28, no. 14 (2023): 5369.
    View at Publisher | View at Google Scholar
11. Zhang, Zheng Fan, Yu Xi, Si Tian Wang, Li Yun Zheng, Ya Qi, Shuang Shuang Guo, and Bin Ying Ding. “Effects of Chinese gallnut tannic acid on growth performance, blood parameters, antioxidative status, intestinal histomorphology, and cecal mi-crobial shedding in broilers challenged with aflatoxin B1.” Jour-nal of Animal Science 100, no. 4 (2022): skac099.
    View at Publisher | View at Google Scholar
12. Tavangar, Pouya, Shahabodin Gharahveysi, Vahid Rezaeipour, and Mehrdad Irani. “Efficacy of phytobiotic and toxin binder feed additives individually or in combination on the growth per-formance, blood biochemical parameters, intestinal morphol-ogy, and microbial population in broiler chickens exposed to aflatoxin B1.” Tropical Animal Health and Production 53, no. 3 (2021): 335.
    View at Publisher | View at Google Scholar
13. Śliżewska, Katarzyna, Bożena Cukrowska, Stefania Smuli-kowska, and Joanna Cielecka-Kuszyk. “The effect of probiot-ic supplementation on performance and the histopathological changes in liver and kidneys in broiler chickens fed diets with aflatoxin B1.” Toxins 11, no. 2 (2019): 112.
    View at Publisher | View at Google Scholar
14. Sarker, Md Touhiduzzaman, Xiaoli Wan, Haiming Yang, and Zhiyue Wang. “Dietary lycopene supplementation could allevi-ate aflatoxin B1 induced intestinal damage through improving immune function and anti-oxidant capacity in broilers.” Animals 11, no. 11 (2021): 3165.
    View at Publisher | View at Google Scholar
15. Hernandez, M and Alagbe, J.O. (2025). Influence of Abrus procatorious crude oil supplementation on the growth perfor-mance, Nutrient digestibility, Ruminal fermentation and Mi-crobial population of Malabari Bucks. International Journal of Global Sustainable Research, 3(7): 527-538.
    View at Publisher | View at Google Scholar
16. Alagbe, J. O., A. A. Eimoga, and O. O. Alagbe. “Growth re-sponse and carcass characteristics of weaner grass cutters fed diets supplemented with Polyalthia longifolia seed oil as a natural growth promoter.” Greener Journal of Agricultural Sci-ences 7, no. 5 (2017): 112-119.
    View at Publisher | View at Google Scholar
17. Morris, Hernández, and Alagbe John Olujimi. “INFLUENCE OF Odontonema Strictum OIL ON THE GROWTH PERFOR-MANCE AND RUMINAL FERMENTATION OF BARBARI BUCKS.” Research in: Agricultural & Veterinary Sciences 9, no. 2 (2025).
    View at Publisher | View at Google Scholar
18. National Research Council, and Subcommittee on Poultry Nu-trition. Nutrient requirements of poultry: 1994. National Acade-mies Press, 1994.
    View at Publisher | View at Google Scholar
19. Rajput, Shahid Ali, Cong Zhang, Yue Feng, Xiao Tian Wei, Mahmoud Mohamed Khalil, Imran Rashid Rajput, Dost Mu-hammad Baloch et al. “Proanthocyanidins alleviates aflatox-inB1-induced oxidative stress and apoptosis through mito-chondrial pathway in the bursa of fabricius of broilers.” Toxins 11, no. 3 (2019): 157.
    View at Publisher | View at Google Scholar
20. Denli, M., J. C. Blandon, M. E. Guynot, S. Salado, and J. F. Pe-rez. “Effects of dietary AflaDetox on performance, serum bio-chemistry, histopathological changes, and aflatoxin residues in broilers exposed to aflatoxin B1.” Poultry Science 88, no. 7 (2009): 1444-1451.
    View at Publisher | View at Google Scholar
21. Chen, Xueping, Muhammad Ishfaq, and Jian Wang. “Effects of Lactobacillus salivarius supplementation on the growth per-formance, liver function, meat quality, immune responses and Salmonella Pullorum infection resistance of broilers challenged with Aflatoxin B1.” Poultry Science 101, no. 3 (2022): 101651.
    View at Publisher | View at Google Scholar
22. Dumrongphol, Yolprapa, Takaya Hirota, Hidehiro Kondo, Takashi Aoki, and Ikuo Hirono. “Identification of novel genes in Japanese flounder (Paralichthys olivaceus) head kidney up-regulated after vaccination with Streptococcus iniae forma-lin-killed cells.” Fish & Shellfish Immunology 26, no. 1 (2009): 197-200.
    View at Publisher | View at Google Scholar
23. Kumar, D. Senthil, Suguna Rao, M. L. Satyanarayana, PG Pradeep Kumar, and N. Anitha. “Amelioration of hepatotoxicity induced by aflatoxin using citrus fruit oil in broilers (Gallus do-mesticus).” Toxicology and Industrial Health 31, no. 11 (2015): 974-981.
    View at Publisher | View at Google Scholar
24. Liu, H. W., K. Li, J. S. Zhao, and W. Deng. “Effects of chestnut tannins on intestinal morphology, barrier function, pro-inflam-matory cytokine expression, microflora and antioxidant capac-ity in heat-stressed broilers.” Journal of Animal Physiology and Animal Nutrition 102, no. 3 (2018): 717-726.
    View at Publisher | View at Google Scholar
25. Rashidi, Nasrin, Ali Khatibjoo, Kamran Taherpour, Mohammad Akbari-Gharaei, and Hassan Shirzadi. “Effects of licorice ex-tract, probiotic, toxin binder and poultry litter biochar on perfor-mance, immune function, blood indices and liver histopatholo-gy of broilers exposed to aflatoxin-B1.” Poultry science 99, no. 11 (2020): 5896-5906.
    View at Publisher | View at Google Scholar
26. El Khoury, Rhoda, Isaura Caceres, Olivier Puel, Sylviane Bail-ly, Ali Atoui, Isabelle P. Oswald, André El Khoury, and Jean-De-nis Bailly. “Identification of the anti-aflatoxinogenic activity of Micromeria graeca and elucidation of its molecular mechanism in Aspergillus flavus.” Toxins 9, no. 3 (2017): 87.
    View at Publisher | View at Google Scholar
27. Solis-Cruz, Bruno, Daniel Hernandez-Patlan, Victor M. Petrone, Karine P. Pontin, Juan D. Latorre, Eric Beyssac, Xochitl Her-nandez-Velasco et al. “Evaluation of cellulosic polymers and curcumin to reduce aflatoxin B1 toxic effects on performance, biochemical, and immunological parameters of broiler chick-ens.” Toxins 11, no. 2 (2019): 121.
    View at Publisher | View at Google Scholar
28. Umaya, Suganthi R., Y. C. Vijayalakshmi, and V. Sejian. “Ex-ploration of plant products and phytochemicals against aflatox-in toxicity in broiler chicken production: Present status.” Toxi-con 200 (2021): 55-68.
    View at Publisher | View at Google Scholar
29. Chen, X., K. Naehrer, and T. J. Applegate. “Interactive effects of dietary protein concentration and aflatoxin B1 on perfor-mance, nutrient digestibility, and gut health in broiler chicks.” Poultry science 95, no. 6 (2016): 1312-1325.
    View at Publisher | View at Google Scholar
30. Chlebicz, Agnieszka, and Katarzyna Śliżewska. “In vitro detox-ification of aflatoxin B1, deoxynivalenol, fumonisins, T-2 toxin and zearalenone by probiotic bacteria from genus Lactobacil-lus and Saccharomyces cerevisiae yeast.” Probiotics and anti-microbial proteins 12, no. 1 (2020): 289-301.
    View at Publisher | View at Google Scholar
31. Choi, Dae-Woon, Sun Young Jung, Jisu Kang, Young-Do Nam, Seong-Il Lim, Ki Tae Kim, and Hee Soon Shin. “Immune-en-hancing effect of nanometric Lactobacillus plantarum nF1 (nLp-nF1) in a mouse model of cyclophosphamide-induced immunosuppression.” (2018): 218-226.
    View at Publisher | View at Google Scholar
32. Omokore, E. O., and J. O. Alagbe. “Efficacy of dried Phyllan-tus amarus leaf meal as an herbal feed additive on the growth performance, haematology and serum biochemistry of growing rabbits.” International Journal of Academic Research and De-velopment 4, no. 3 (2019): 97-104.
    View at Publisher | View at Google Scholar
33. Alagbe, J. O., A. A. Eimoga, and O. O. Alagbe. “Growth re-sponse and carcass characteristics of weaner grass cutters fed diets supplemented with Polyalthia longifolia seed oil as a natural growth promoter.” Greener Journal of Agricultural Sci-ences 7, no. 5 (2017): 112-119.
    View at Publisher | View at Google Scholar
34. Alagbe, J. O. “Growth performance and blood parameters of weaner pigs fed diets supplemented with turmeric powder.” En- ergy (ME kcal/kg) 2433, no. 2437.5 (2017): 2439-9.
    View at Publisher | View at Google Scholar