Iranian Journal of Veterinary Science and Technology

Iranian Journal of Veterinary Science and Technology

Hematological Parameters and Splenic Histopathology in Offspring of Wistar Rats Exposed to Prenatal Stress and Moringa oleifera

Document Type : Research Article

Authors
Odochi Ogbu Chukwu 1
Anthony C.U. Ezimah 2
Nwaeze Konyefom 2
Grace Ottah-umahi 2
Sylvester Ibekailo 2
1 Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, David Umahi Federal University of Health Sciences, Ebonyi State & Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, Alex Ekwueme Federal University, Ndufu-Alike. Abakaliki, Ebonyi State & Physiology Unit, College of Health Sciences, Evangel University, Akaeze, Nigeria.
2 Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, Alex Ekwueme Federal University, Ndufu-Alike. Abakaliki, Ebonyi, State, Nigeria.
10.22067/ijvst.2025.91007.1447
Abstract
This study investigates the effects of prenatal exposure to Moringa oleifera leaf extract (MoLE) and chronic unpredictable stress (CUS) on the immune development and splenic histopathology of offspring in rat model. Twenty-five pregnant Albino-Wistar rats were divided into five groups: a control group, two MoLE-treated groups (5 mg/kg and 10 mg/kg), and two CUS-exposed MoLE-treated groups (5 mg/kg and 10 mg/kg). MoLE was administered via gavage from gestation day 8 to 21. Hematological parameters and splenic histopathology were assessed in offspring at puberty. Results showed that MoLE exposure affected blood cell counts and splenic histology. Specifically, MoLE treatment resulted in decreased monocyte and granulocyte counts, while lymphocyte levels remained unchanged. The high-dose MoLE group exhibited increased mean platelet volume (MPV) and platelet distribution width (PDW). Severe histopathological changes, including fibrosis and hemorrhage, were observed in the CUS-exposed groups. These findings suggest that prenatal MoLE and CUS exposure influence immune development.
Keywords
Prenatal Stress
Hematotoxicity
Immunity
Blood
Moringa oleifera
Subjects

Abbreviations

AE-FUNAI: Alex Ekwueme Federal University Ndufu-Alike

ANOVA: Analysis of Variance

CUS: Chronic Unpredictable Stress

CV: Coefficient of Variation

EDTA: Ethylenediaminetetraacetic Acid

GC–MS: Gas Chromatography-Mass Spectrometry

Introduction

Chronic unpredictable stress (CUS) during pregnancy represents a significant public health concern due to its potential to disrupt fetal development, and impacting both physiological and behavioral outcomes in offspring [ 1 ]. Prenatal stress, especially in the form of CUS, has been associated with alterations in hematological profiles, such as changes in white blood cell (WBC) counts and platelet function, which could have lasting effects on immune development and blood clotting processes [ 1 - 3 ]. These hematological alterations may increase offspring susceptibility to infections and other health complications [ 4 ]. Thus, identifying potential interventions to mitigate these adverse effects is crucial for promoting healthy offspring development.

Moringa oleifera, commonly referred to as the “drumstick tree,” is renowned for its health-promoting properties. This drought-resistant tree is cultivated in tropical and subtropical regions and is particularly valued for its leaves, which are rich in essential nutrients, vitamins, minerals, and bioactive compounds, including flavonoids, glucosinolates, and phenolic acids [ 5 , 6 ]. These compounds are believed to contribute to the plant’s antioxidant, anti-inflammatory, and immunomodulatory effects [ 7 ].

Although the therapeutic potential of MoLE, has been increasingly documented, its effects during sensitive developmental periods, particularly during pregnancy, remain insufficiently characterized. Furthermore, the combined influence of MoLE and CUS exposure on offspring development, especially regarding hematological parameters and splenic health, has not been fully elucidated. This study aims to bridge this gap by examining the effects of MoLE on hematological markers and splenic histopathology in offspring exposed to prenatal CUS, thus contributing valuable insights into maternal health and offspring development.

Results

The majour compounds identified in the GC-MS analysis of MOLE are known for their antioxidant and anti-inflammatory activities [ 8 ](Table 1).

S/No Name of Compound Mol. formular Mol. Wt(g) RT (min) % TIC Structure Activity
1 1-Propanol, 3,3'-oxybis- C6H14O3 134. 3.045 1.521 Humectants (Food additive/Moisturizer)
2 1-Propanamine, 3-propoxy- C5H12NO 117 3.327 0.803 Textile resins, Drugs, Pesticides
3 2-Pentene, 2-methyl- C6H12 84 3.778 0.593 Photochemical and ozonolysis studies
4 Pyridine C5H5N 79 4.285 4.763 Drugs, Vitamins, Food flavorings, Pesticides,
5 2-Pentanone, 5-hydroxy- C5H10O2 102 4.820 0.630 Anti-malarial drugs, Vitamin B1
6 5-Hexen-2-ol, 5-methyl- C7H14O 114 5.102 0.887 Natural substances and Extractives
7 1,3-Propanediamine, N-(1-methylethyl)- C6H16N2 116 5.440 0.546 Useful research chemical compound
8 1,4-Butanediamine, N,N'-diethyl- C8H20N2 144 5.553 0.183 Unidentified
9 Hexanoic acid, methyl ester C7H14O2 130 5.722 0.843 Flavouring agents
10 Cyclotetrasiloxane, octamethyl- C8H24O4Si4 296 6.031 0.468 Pharmaceuticals, Polymers, Hair/Skin care products, Antiperspirants and Deodorants, Lubricants, Sealants, Adhesives, Waxes and Coating.
11 1,2,3-Trimethyldiaziridine C4H10N2 86 6.285 0.515 Unidentified
12 2-Hexyn-1-ol C6H10O 98 6.426 0.212 Flavour and fragrance
13 Heptanoic acid, methyl ester C8H16O2 144 6.595 1.204 Human Metabolite, Flavouring agents, Fragrance,
14 1-Heptene, 3-methyl- C8H16 112 6.905 0.608 Hydrocarbon
15 1-Fluorononane C9H19F 146 7.158 0.459 Unidentified
16 Octanoic acid, methyl ester C9H18O2 158 7.440 2.299 Metabolite
17 Erythritol C4H10O4 122 7.834 6.842 Food additive and Sugar substitutes
18 2-Mercaptopropanoic acid C3H6O2S 106 8.313 2.240 Flavour and Fragrance agents
19 Triethylene glycol C6H14O4 150 9.243 1.079 Pesticides, Fragrance, Humectant, Disinfectant, Plasticizer for vinyl polymers
20 Decanoic acid, methyl ester C11H22O2 186 9.440 1.750 Biodiesel surrogate
21 Benzene,2-methoxy-1,3,4-trimethyl C10H14O 150 10.849 2.298 unidentified
22 Ethylene, 1,2-dichloro-, (Z)- C2H2Cl2 97 10.894 0.200 Pharmacology, Refrigerant, Degreaser, Adhesives, Lacquers, oils, and Resins
23 10-Undecenoic acid, methyl ester C12H22O2 198 11.102 0.275 Flavouring agents
24 Trisiloxane, 1,1,1,5,5,5-hexamethyl-3,3-bis[(trimethylsilyl)oxy]- C12H36O4Si5 385 11.581 0.360 Paints, Coatings, and Cosmetics, including some Personal care products
25 7-Hexadecenal, (Z)- C16H30O 238 12.539 0.016 Derivative of essential oils with potential antibacterial activities.
26 1-Octanol, 2-butyl- C12H26O 186 12.623 0.015 Human metabolite, Humectant
27 3,8-Dioxatricyclo[5.1.0.0(2,4)]octane, 4-ethenyl- C8H10O2 138 12.905 0.003 Undefined
28 Dodecanoic acid, methyl ester C13H26O2 214 13.356 6.008 Therapeutic uses, Flavouring agents
29 1-Decanol, 2-hexyl- C16H34O 242 14.342 0.497 Fungicidal properties, Inhibitor (Candida glabrata), suggesting useful for treating Skin cancer
30 Myristyl stearate C32H64O2 481 14.623 0.181 Skin conditioning and Deodorant
31 2-Tridecenal, (E)- C13H24O 196 14.877 0.219 Flavour and Fragrance agents
32 Cyclotetradecane C14H28 196 15.468 1.427 Plant metabolite and a human metabolite
33 Methyl tetradecanoate C15H30O2 242 15.891 4.899 Plant metabolite, Flavouring agent and a Fragrance.
34 2-Piperidinone, N-[4-bromo-n-butyl]- C9H16BrNO 234 16.426 1.766 Antimicrobial
35 Tetradecanal C14H28O 212 16.736 1.853 Human metabolite, Flavouring agent and Fragrance
36 Ethanol, 2-(octadecyloxy)- C20H42O2 314 16.877 1.420 Surfactant
37 Hexadecanoic acid, methyl ester C17H34O2 270 17.384 26.182 Metabolite
38 6-Ethoxy-6-methyl-2-cyclohexenone C9H14O2 154 17.778 3.234 Flavouring agent
39 n-Dodecyl methacrylate C16H30O2 254 18.004 3.814 Drug (Clinical trials), Metabolites, Fragrance, Pesticides etc.
40 9-Octadecenoic acid (Z)-, methyl ester C19H36O2 296 18.567 4.770 Flavouring agents and Fragrance
41 Methyl stearate C19H38O2 298 18.736 12.113 Metabolites and Flavouring agents
Table 1.GC-MS analysis of MOLE; An exempt from “GC–MS analysis of Moringa oleifera leaf extract and effects of administration on histology of reproductive organs and liver of female rats exposed to chronic unpredictable stress” by Chukwu et al., 2024 [ 10 ]

White Blood Cell and Differential Counts

Prenatal exposure to Moringa oleifera leaf extract (MoLE) and/or chronic unpredictable stress (CUS) resulted in significant alteration in leukocyte profiles in the offspring (Table 2). Total white blood cell (WBC) counts were significantly elevated in the High dose MoLE group (17.42 ± 1.78 ×109/L) and the CUS + High dose MoLE group (15.32 ± 3.39 ×109/L) when compared with the Control group (5.40 ± 0.22 ×109/L, p < 0.05). Lymphocyte counts exhibited a similar pattern, with significant increases observed in the High-dose MoLE and CUS + High dose MoLE groups (p < 0.05).

Variable (Units) Control Low-dose MoLE High-dose MoLE CUS + Low-dose MoLE CUS + High-dose MoLE
WBC (×109/L) 5.40±0.22a 10.87±1.64b 17.42±1.78c 7.32±2.15ab 15.32±3.39c
Lymphocytes (×109/L) 4.43±0.07a 7.93±0.77b 13.03±1.54c 6.90±2.07ab 11.86±2.49c
Monocytes (×109/L) 0.51±0.10a 0.54±0.19a 0.95±0.11ab 0.30±0.06a 0.63±0.18ab
Granulocytes (×109/L) 0.46±0.15a 2.40±0.71b 3.45±0.15c 0.12±0.03a 2.83±0.72c
Platelet Indices
PLT (×109/L) 1087.3±115.9a 355.7±51.0b 312.3±40.1b 1445.0±101.9c 316.0±17.2b
MPV (fL) 7.07±0.03a 7.27±0.12ab 7.77±0.23c 6.83±0.09a 7.60±0.15bc
PDW-CV (%) 13.97±0.13a 14.00±0.25a 15.07±0.12b 13.83±0.09a 14.10±0.17ab
PDW-SD (fL) 9.77±0.19a 10.53±0.45ab 12.30±0.44c 9.43±0.03a 11.50±0.42bc
PCT (mL/L) 7.71±0.86a 2.59±0.35b 2.44±0.37b 5.59±2.80ab 2.41±0.13b
P-LCR (%) 14.60±0.26a 15.33±0.94a 18.77±1.27b 12.60±0.65a 16.77±0.98b
Values are expressed as Mean ± SEM. Within each row, means that do not share the same superscript letter differ significantly (p < 0.05). Superscripts with the same letter indicate no statistically significant difference between the corresponding groups.
 Table 2.Hematological Parameters (White Blood Cell, Differential Count, and Platelet Indices) of Offspring of Dams Exposed to CUS and MoLE

Granulocyte counts were significantly higher in the High dose MoLE (3.45 ± 0.15 ×109/L) and the CUS + High dose MoLE groups (2.83 ± 0.72 ×109/L) compared to Control (0.46 ± 0.15 ×109/L, p < 0.05). In contrast, no significant changes were observed in monocyte counts across groups (p > 0.05). These findings suggest a stress induced inflammatory response that was partially modulated by MoLE supplementation in a dose dependent manner.

Platelet Counts and Indices

Platelet profiles also significantly affected by prenatal treatments (Table 2). Offspring from High dose MoLE and both CUS exposed groups exhibited a sharp reduction in platelet counts compared to Control, showing a decrease of over 70% (p < 0.05). In contrast, CUS + Low-dose MoLE group exhibited a significant increase in platelet count (1445.0±101.9 ×109/L, p < 0.05) compared to Control.

Analysis of platelet indices revealed that mean platelet volume (MPV) and platelet distribution width (PDW-SD and PDW-CV) were significantly elevated only in the High-dose MoLE group (p < 0.05). No significant differences in MPV or PDW values were observed in the CUS + Low-dose MoLE group compared with Control, indicating preservation of platelet morphology under low-dose supplementation.

Histopathological Findings

Representative photomicrographs of spleen sections are shown in Figure 1 (Panels A–E). Spleens from Control animals exhibited normal histological architecture, characterized by well-demarcated white pulp and red pulp regions (Figure 1A). Low-dose MoLE-treated offspring displayed mild splenic tissue degeneration accompanied by moderate inflammatory infiltration and focal ground-glass changes (Figure 1B). In contrast, High-dose MoLE offspring showed mild to moderate fibrotic changes with occasional hemorrhage (Figure 1C). The most severe histopathological changes were observed in the CUS + Low-dose MoLE group, characterized by marked tissue degeneration, extensive fibrosis, and hemorrhagic red pulp (Figure 1D). The CUS + High-dose MoLE group exhibited moderate tissue degeneration with focal fibrosis but less extensive damage compared to the CUS + Low-dose MoLE group (Figure 1E), suggesting a partial protective effect of higher MoLE supplementation under stress exposure. Semi-quantitative lesion scoring (Table 3), confirmed these histological observations: fibrosis and hemorrhage were significantly more severe in the CUS + Low-dose MoLE group compared to Control (p < 0.05). In contrast, co-treatment with High-dose MoLE attenuated lesion severity under CUS conditions (p < 0.05).

Group Fibrosis (0–3) Hemorrhage (0–3) Inflammatory Infiltrate (0–3) Ground Glass Areas (0–3) Total Lesion Score (0-12)
Control 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00a
Low-dose MoLE 1.00 ± 0.00 0.00 ± 0.00 2.00 ± 0.00 2.00 ± 0.00 5.00 ± 0.00b
High-dose MoLE 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 0.00 ± 0.00 3.00 ± 0.00b
CUS + Low-dose MoLE 3.00 ± 0.00 3.00 ± 0.00 2.00 ± 0.00 0.00 ± 0.00 8.00 ± 0.00c
CUS + High-dose MoLE 2.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 0.00 ± 0.00 4.00 ± 0.00b
Values are mean ± SEM (n = 3 animals per group). 0 = absent, 1 = mild, 2 = moderate, 3 = severe. Superscripts within the Total Lesion Score column indicate statistically distinct groups (p < 0.05). Groups sharing the same superscript are not significantly different from each other.
aControl, bLow-dose MoLE, High-dose MoLE, and CUS + High-dose MoLE (not significantly different from one another), cCUS + Low-dose MoLE (significantly higher than all other groups)
 Table 3.Semi-quantitative Scoring of Splenic Lesions in Offspring

Figure 1. Representative photomicrographs of spleen sections (H&E staining) showing:

  • (B) Low-dose MoLE: Mild degeneration with inflammatory infiltration;
  • (C) High-dose MoLE: Mild fibrosis and hemorrhage;
  • (D) CUS + Low-dose MoLE: Severe degeneration, fibrosis, and hemorrhagic red pulp;
  • (E) CUS + High-dose MoLE: Moderate degeneration with focal fibrosis.

Figure 1. A: Photomicrograph of Control section of the spleen (x150) (H/E) shows normal spleen architecture with Red Pulp (RP) and White Pulp (WP), with Central Spleenic Artery. B: Photomicrograph of Low-dose MoLE section of the Spleen (x150) (H/E) shows Mild Degeneration with moderate Focal Area of Ground Glass (GG) and moderate Infilteration (IIC). C: Photomicrograph of High-dose MoLE section of the Spleen (x150) (H/E) shows mild to moderate degeneration with mild fibrosis (F) and hemorrhage (H). D: Photomicrograph of CUS + Low-dose MoLE section of the Spleen (x100) (H/E) shows severe degeneration of the Spleenic Tissue with severe Fibrosis (F) and Hemorrhagic (H) Red Pulp. E: Photomicrograph of CUS + High-dose MoLE section of the Spleen (x100) (H/E) shows moderate Degeneration with mild Focal Area of Fibrosis (F).

Declaration of Generative AI and AI-Assisted Technologies in the Writing Process

During manuscript preparation, a generative AI tool (ChatGPT, OpenAI) was used only as a supportive language aid to improve sentence structure, grammar, and clarity in selected sections of the manuscript. The tool was not used to generate scientific hypotheses, experimental design, data, statistical analyses, figures, or interpretations. All content was critically reviewed, edited, and finalized by the authors, who take full responsibility for the originality, accuracy, and integrity of the work.

Ethical Approval

All experimental procedures were conducted in accordance with the guidelines for the care and use of laboratory animals and were approved by the Faculty of Basic Medical Sciences Research Ethics Committee, Alex Ekwueme Federal University Ndufu-Alike, Ebonyi State, Nigeria (Approval Code: FBMS/EC/AE/1983).

Plant Collection, Identification, and Extraction

Fresh Moringa oleifera leaves were collected in the early morning from a cultivated garden in Abakaliki, Ebonyi State. Botanical authentication was confirmed at the Herbarium Unit, Department of Biology, Alex Ekwueme Federal University Ndufu-Alike (AE-FUNAI). Leaves were thoroughly washed, air-dried at room temperature for seven days, and pulverized into a coarse powder using an electric blender (Model MS-233, China). Extraction was performed using methanol according to standardized procedures [ 5 , 6 ]. The filtrate was concentrated at 40°C under reduced pressure to yield a dark-green paste, which was stored at 4°C until use.

Phytochemical characterization of the extract was performed using gas chromatography–mass spectrometry (GC–MS) as previously described by Chukwu et al. [ 8 ]. Analysis was performed using an Agilent GC–MS system equipped with an HP-5MS capillary column (30 m × 0.25 mm, 0.25 µm). The oven temperature was programmed from 60°C (2 min hold) to 280°C at 10°C/min. Helium was used as the carrier gas at a flow rate of 1.0 mL/min. Major compounds identified are known for antioxidant and anti-inflammatory activity [ 8 ] (Table 1).

Experimental Animals and Housing Conditions

Twenty-five mature, nulliparous, virgin female Albino-Wistar rats (weighing 150–180 g) were obtained from the Animal House, AE-FUNAI. Animals were housed in well-ventilated polypropylene cages under standard laboratory conditions, including 12h light/dark cycle, temperature of 23±2°C, and humidity of 50-60%. Rats had free access to standard rat chow (Vital feed®, Nigeria) and tap water. Animals were acclimatized for two weeks before mating.

Estrous Cycle Monitoring and Mating

Estrous cycle monitoring was performed by vaginal cytology using light microscopy following established protocols [ 9 ]. Females with two consecutive regular four-day cycles were considered for mating. During the proestrus phase, identified by the predominance of nucleated epithelial cells and absence of cornified cells, females were co-housed overnight with proven male breeders (1:2 ratio). The presence of spermatozoa in morning smears confirmed successful mating and was designated as gestational day (GD) 1 [ 10 ].

Experimental Design and MoLE Administration

Pregnant rats were randomly assigned (n=5 per group) to the following treatment groups:

  • • Control: Standard diet and water ad libitum.
  • • Low-dose MoLE: MoLE 5 mg/kg/day via oral gavage from GD 8–21.
  • • High-dose MoLE: MoLE 10 mg/kg/day via oral gavage from GD 8–21.
  • • CUS + Low-dose MoLE: CUS exposure plus 5 mg/kg/day MoLE from GD 8–21.
  • • CUS + High-dose MoLE: CUS exposure plus 10 mg/kg/day MoLE from GD 8–21.

Doses were freshly prepared in distilled water daily and administered in volumes not exceeding 1 mL/100 g body weight to avoid gastric discomfort [ 8 ]. The gestational window GD 8-21 was selected to reflect the late second to third trimester in human pregnancy, a critical period for immune and hematopoietic development [ 11 , 12 ].

Chronic Unpredictable Stress (CUS) Protocol

Animals in CUS groups were subjected to a validated CUS protocol [ 13 ] consisting of the following randomly applied stressors:

  • • Wet bedding (300 mL water mixed with sawdust).
  • • Cage tilting at 45° for 6 hours.
  • • Overnight food deprivation.
  • • Psychological stress by exposure to a caged cat.
  • • Sleep deprivation using a pedestal in shallow water.
  • • Restraint stress in 50 mL plastic tubes for 2-hour intervals.
  • • Continuous overnight light exposure.
  • • Social isolation for 6-12 hours.

Stressors were rotated daily in an unpredictable order from GD 8–21 to mimic human psychosocial stress [ 13 , 14 ].

Sample Collection and Litter Effect Consideration

At the onset of puberty (postnatal day 21 onward), five offspring per treatment group were sampled for hematological analysis, with at least one pup selected from each dam (total 5 dams per group). To minimize litter effect bias and maintain balanced sex representation, pups were randomly selected across litters, ensuring that no more than one male and one female pup per dam contributed to the dataset. This approach ensured equal representation of each dam while avoiding over-representation of littermates.

Although individual pup measurements were recorded, the dam (litter) was treated as the experimental unit (n = 5 dams per group) for all statistical analyses, in accordance with internationally accepted developmental toxicology guidelines [ 12 , 16 ], thereby preventing pseudo-replication. Approximately 2 mL of blood was collected via retro-orbital venous puncture under light isoflurane anesthesia into EDTA-coated tubes and processed within 3 hours at 4°C to preserve cell morphology and ensure accurate hematological profiling.

Hematological Analysis

A calibrated automated hematology analyzer (Mindray BC-2800, Shenzhen, China) was used to determine:

  • • Total WBC counts (×109/L).
  • • Differential counts: Lymphocytes (Lym), monocytes (MID), and granulocytes (Gran), reported as absolute counts and percentages.
  • • Platelet parameters: Platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW-CV, PDW-SD), plateletcrit (PCT), and platelet-large cell ratio (P-LCR) [ 16 , 17 ].

Histological Studies

Following blood collection, animals were humanely euthanized using sodium pentobarbital (150 mg/kg, i.p.). Spleens were excised, trimmed of fat, and fixed in 10% neutral-buffered formalin for 24-48 hours. Standard tissue processing, paraffin embedding, and hematoxylin-eosin (H&E) staining were performed [ 18 ]. Sections were examined under a light microscope (×100–150 magnification). Histopathological changes, including fibrosis, hemorrhage, inflammatory infiltration, ground-glass appearance; were semi-quantitatively graded on a scale of 0-3 by two independent blinded observers. Any scoring discrepancies were resolved by consensus [ 19 ].

Statistical Analysis

All data are presented as mean ± standard error of the mean (SEM). The dam was considered as the experimental unit (n = 5 per group), to avoid pseudo-replication. When more than one pup per dam was sampled, values were averaged to obtain a single representative data point per dam. Hematological parameters were analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test for multiple comparisons. Non-parametric data from semi-quantitative splenic lesion scoring were analyzed using the Kruskal–Wallis test followed by Dunn’s multiple comparisons. A p-value <0.05 was considered statistically significant. Statistical analyses were performed using GraphPad Prism (Version 9.0, GraphPad Software, San Diego, CA, USA).

Authors' Contributions

O.O.C., A.C.U.E, conceived, planned and carried out the experiments, O.O.C., S.N.I.and N.G.K carried out data collection and analysis. O.O.C. wrote the first draft of the manuscript, and all authors provided critical feedback and helped shape the research, analysis and manuscript.

Acknowledgements

The authors express their gratitude to the Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, AE-FUNAI, Ebonyi State, for their support during this study.

Conflict of interest

The authors declare that there is no conflict of interest.

Abbreviations-Cont'd

GD: Gestational Day

Gran: Granulocytes

HPA: Hypothalamic–Pituitary–Adrenal (Axis)

i.p.: Intraperitoneal

Lym: Lymphocytes

MID: Monocytes/Intermediate Cells

MoLE: Moringa oleifera Leaf Extract

MPV: Mean Platelet Volume

PCT: Plateletcrit

PDW: Platelet Distribution Width

PDW-CV: Platelet Distribution Width Coefficient of Variation

PDW-SD: Platelet Distribution Width Standard Deviation

P-LCR: Platelet Large Cell Ratio

PLT: Platelet Count

SD: Standard Deviation; SEM: Standard Error of the Mean

BC: White Blood Cell

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  • Receive Date 20 December 2024
  • Revise Date 08 October 2025
  • Accept Date 28 October 2025