Impact of Zingiber officinale on Testicular Morphometry, Sperm Quality, and Hormonal Profiles in Alcohol-Induced Toxicity

Impact of Zingiber officinale on Testicular Morphometry, Sperm Quality, and Hormonal Profiles in Alcohol-Induced Toxicity | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Impact of Zingiber officinale on Testicular Morphometry, Sperm Quality, and Hormonal Profiles in Alcohol-Induced Toxicity Collins Nduka Esomchi, Joy Onyinyechi Nwofia, Favour Kyrian Nwoke, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5784131/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Alcohol consumption is known to induce reproductive toxicity, leading to adverse effects on testicular morphology, sperm quality, and DNA integrity in males. Zingiber officinale (ginger), known for its antioxidant and anti-inflammatory properties, may counteract these effects. This study aimed to investigate the impact of ginger on testicular morphometry, sperm quality, and hormonal profiles in alcohol-induced toxicity. 30 male Wistar rats were divided into six groups (n=5/group). Group A (control) received normal saline. Group B was exposed to 40% alcohol (3.50 g/kg body weight) from Days 15–28. Group C received ginger (750 mg/kg) during the same period. Groups D, E, and F were treated with alcohol for 14 days, followed by low (250 mg/kg), medium (500 mg/kg), and high (750 mg/kg) doses of ginger, respectively, from Days 15–28. The study evaluated changes in body and testicular morphometry, antioxidant enzyme and hormonal changes. Semen analysis included sperm motility, count, and morphology, while sperm chromatin/DNA integrity was assessed using Aniline Blue and Toluidine Blue staining. Results: Alcohol exposure (Group B) significantly reduced testicular weight, sperm motility, and chromatin integrity. Ginger-treated groups (C, D, and E) showed marked improvements, with Group C outperforming the control group (A) in sperm motility, antioxidant levels, and hormones. However, Group F showed distorted results similar to Group B, suggesting high-dose toxicity. Conclusion : Ginger improves reproductive health and mitigates alcohol-induced toxicity in a dose-dependent manner. Moderate doses show optimal benefits, while high doses may be detrimental. These findings support ginger’s potential as a natural therapeutic agent for reproductive health. Alcohol-induced toxicity Antioxidant Ginger Semen quality Testicular morphometry Zingiber officinale Figures Figure 1 Figure 2 Figure 3 1. Introduction Infertility known as the inability to conceive after months of regular, unprotected sexual intercourse for at least 12 months is a significant health matter [ 1 ]. On different scales, it is known as a medical illness because it affects more than one person instead of a person [ 2 ]. Globally, infertility affects about 15% of couples of reproductive age [ 3 ]. Males account for about 50% of total infertility cases worldwide, the prevalence is said to increase at 0.3% annually leading to an uneven geographical distribution ranging from 20 to 70% [ 4 ]. Nonetheless, the rates of male infertility are underreported due to different factors such as cultural, societal, religious and patriarchal influences that hinder precise sampling and analysis [ 5 ]. Male infertility includes any health condition that hinders the chances of conception. It can result from abnormal sperm function or obstructions preventing ejaculation [ 6 ]. Numerous factors are reported to influence male infertility which include anatomo-pathophysiological factors, environmental factors, the process of ageing and lifestyle factors [ 7 ]. Today, lifestyle-related factors play a significant role in causing male infertility globally [ 8 ]. An example of these lifestyle-related factors is alcohol consumption which has gained considerable attention due to its global prevalence and modifiable nature [ 9 ]. Alcohol is considered one of the most prevalent dietary factors which people are exposed to and very commonplace in many societies, with almost 60% of the global population aged 15 years and over reported to have consumed alcoholic drinks in a single year [ 10 ]. The consumption of alcoholic beverages has been a part of the socio-cultural heritage of most populations since ancient times [ 11 ]. Indeed, alcohol has been viewed as an integral part of a meal, and in some cases, as a remedy for infectious diseases or even as a cleaning agent [ 12 ]. Alcohol consumption poses a significant threat to male reproductive health, affecting millions worldwide and impacting testicular structure and function through oxidative stress mechanisms [ 3 ]. Chronic alcohol exposure leads to histological, hormonal, and biochemical alterations, often culminating in reduced semen quality, including impaired sperm concentration and motility—key markers of male infertility [ 12 ]. Since alcohol causes these distortions via the increase of reactive oxygen species (ROS), many therapeutic approaches focus on synthetic or natural antioxidants derived from natural sources to mitigate the damage caused by an elevation in ROS [ 13 , 14 ]. An example of these natural antioxidants is Zingiber officinale commonly known as ginger. The ginger rhizome is utilised for its aromatic smell and strong taste [ 15 ]. Moreover, ginger is extensively employed in folk medicine for its numerous health benefits in treating various diseases, including chronic conditions such as cancer, diabetes, Alzheimer's, ulcers, cardiovascular disease as well as depression [ 16 ]. The positive impact of ginger on these diseases primarily stems from its antioxidant, antimicrobial, and anti-inflammatory properties [ 17 ]. In different protective studies, ginger has been shown to overcome the reproductive toxicity of cyclophosphamide [ 18 ], gentamicin [ 19 ], sodium arsenite [ 20 ] and ethanol [ 21 ] and increase sperm counts, viability, motility, and hormones and improve testicular architecture. Despite these promising findings, the curative potential on testicular morphometry, sperm quality, and hormonal profiles in alcohol-induced toxicity remains underexplored, hence, this study. 2. Methodology 2.1 Materials The materials for this study include Wistar rats, alcohol solution, Z. officinale extract, assay kits, laboratory equipment etc. 2.2 Study Area This study was carried out in the animal unit of the Anatomy department at Alex Ekwueme Federal University Ndufu Alike Ikwo (AE-FUNAI), Ebonyi State, Nigeria. Ethical approval was sought from the Research and Ethical Committee of the Faculty of Basic Medical Sciences with the code AE-FUNAI/FBMS/EAHC/24/006. 2.3 Samples Collection, Identification and Preparations Fresh samples of ginger tuber were purchased at Ogbe Hausa in Abakaliki Local Government Area, Ebonyi State, Nigeria. The samples were identified in the Applied Biology Department at Ebonyi State University Abakaliki, Nigeria with the voucher number EB/06006. 3g of ginger was shredded into small pieces, dried at room temperature, and then mechanically milled to a fine powder. The obtained solution was dried with a water bath at 40°C, and 2.5g, 5g and 7.5g of the extract were mixed with 30ml, 50ml and 70ml of H 2 O for the low dose, medium dose and high dose treatment, respectively. Otunola & Afolayan [ 22 ] report that 5000 mg/kg is not toxic to Wistar rats. 2.4 Alcohol Preparation The alcohol (99.7–100%vv GPR absolute ethanol) produced by NAFCO Scientific Supplies Limited, Surulere Lagos, Nigeria with Product Number 28304 7k was purchased which was used as the toxicant for this experiment. According to the study Biney et al. [ 23 ], 42% had no mortality in rats. 2.5 Animal Care Thirty (30) male Wistar rats randomly divided into six (6) groups (Groups A-F), each comprising five (5) randomised rats were obtained from the animal house at AE-FUNAI and were kept at room temperature (20–22°C). The animals were housed in well-ventilated cages with suitable environmental conditions. During their acclimatisation process, which lasted for two weeks at the animal house, the rats were provided with standard feed only. Group A served as the normal control group and were administered sterile water throughout the experiment; Group B served as the negative control receiving only 3.50g/kg.bw.d of 40% alcohol for 14 days; Group C served as the positive control and was given a high dose of ginger extract (750 mg/kg.bw) for 14 days; Group D served as an experimental group and was induced with alcohol for 14 days, then treated with a low dose of ginger extracts (250 mg/kg.bw) for another 14 days; Group E as an experimental group was induced with alcohol for 14 days and treated with a medium dose of ginger extracts (500 mg/kg.bw) for 14 days; and, Group F as an experimental group was induced with alcohol for 14 days and treated with a high dose of ginger extracts for 14 days. 2.6 Animal Sacrifice and Sample Collection On the 28th day of the experiment, animals were weighed and sacrificed using cervical dislocation. Blood samples were collected by heart puncture and were centrifuged (3000rpm for 15 min) to obtain sera and they were then separated and stored at -80ºC for later hormonal assays. After blood collection, testicular parameters (weight, size and gonadosomatic/testes index) were performed. The left testis was manually homogenized and centrifuged at 3000rpm for 10 min in a cold phosphate buffer (pH 7.4, 0.1M). The obtained supernatant was used to evaluate the level of hormones and antioxidant enzyme activity. 2.7 Testes Morphometry 2.7.1 Testis Weight The weight of each testis was recorded immediately after extraction using an electronic scale. 2.7.2 Testis Size A sliding digital Vernier calliper was used to measure the width and length of each testis. The testis size was then calculated using the spheroid formula: Testis size = width 2 × length × 0.523 (mm) 2.7.3 Gonadosomatic/Testes Index The final body and testis weights were used to calculate the gonadosomatic index using the formula previously reported by Ukoha et al. [ 24 ]. $$\:\text{G}\text{o}\text{n}\text{a}\text{d}\text{o}\text{s}\text{o}\text{m}\text{a}\text{t}\text{i}\text{c}\:\text{i}\text{n}\text{d}\text{e}\text{x}\:=\:\frac{\text{T}\text{e}\text{s}\text{t}\text{i}\text{s}\:\text{w}\text{e}\text{i}\text{g}\text{h}\text{t}\:}{\text{B}\text{o}\text{d}\text{y}\:\text{w}\text{e}\text{i}\text{g}\text{h}\text{t}\:}\:X\:100\left(\%\right)$$ 2.8 Biochemical Assay 2.8.1 Hormonal Assay The serum levels of reproductive hormones—luteinizing hormone (LH), follicle-stimulating hormone (FSH), and inhibin B (INHB)—were measured using enzyme-linked immunosorbent assay (ELISA) kits. The ELISA kits for LH and FSH were obtained from NIADDK, NIH (USA), while the kit for INHB was sourced from Diagnostic Systems Laboratories (DSL-10-84100i; Webster, TX, USA), following the manufacturer’s protocols, as reported by Famurewa et al . [ 25 ]. 2.8.2 Glutathione (GSH) Level Testicular activities of antioxidant enzyme GSH were analysed in homogenate supernatant using commercial rat ELISA kits. 2.8.3 Semen Analysis The sperm was deposited on a clean, dry glass slide and mixed with an equal volume of 1% NaHCO3 buffered Tyrodes Lactate solution to assess the percentage of sperm motility, count, and morphology, following the procedures outlined by Ovie et al . [ 26 ]. 2.8.4 Sperm Chromatin Evaluation Standard cytochemical methods, incorporating aniline blue (AB) and toluidine blue (TB), were employed to evaluate chromatin condensation and DNA integrity. AB was selectively utilised to stain lysine-rich histones. Air-dried smears obtained from washed semen samples were positioned in 0.2 M phosphate buffer (pH 7.2) containing 3% buffered glutaraldehyde for 30 minutes at room temperature. Subsequently, each smear underwent staining in 4% acetic acid (pH 3.5) with a 5% aqueous solution of AB for 7 minutes. During the light microscopic evaluation, a meticulous count of 200 spermatozoa was conducted across various sections of each slide, utilizing a ×100 eyepiece magnification [ 27 ]. Sperm heads stained pale blue/colourless and dark blue were considered normal (AB-) and abnormal sperm (AB+), respectively. On the other hand, TB served as a metachromatic dye, offering insight into nuclear chromatin condensation and the quality and quantity of DNA fragmentation in sperm. Air-dried sperm smears were fixed using a mixture of 96% ethanol and acetone (1:1) for 30 minutes at a temperature of 4°C. Subsequently, the slides underwent a 5-minute incubation in 0.1 N HCl at 4°C, followed by thorough washing with distilled water three times for 2 minutes each. Finally, staining took place for 10 minutes at room temperature using 0.05% TB in 50% citrate phosphate. In the evaluation process, a minimum of 200 spermatozoa were counted in each sample using light microscopy with a ×100 eyepiece magnification [ 27 ]. Normal sperm is pale blue and abnormal sperm is dark blue or violet purple. For each sample, the normal (TB-) and abnormal (TB+) spermatozoa were reported as percentages. 2.9 Data Analysis Data were subjected to analysis of variance using GraphPad Prism version 8 and presented as Means ± SD. Group means of parametric data were compared using a one-way analysis of variance, followed by Turkey’s post hoc test. p < 0.05 was considered statistically significant. 3. Results 3.1 Body Weight Analysis Table 1 Body weight analysis among the experimental groups. Groups Initial Weight (g) Final Weight (g) Weight change (g) A 113.20 ± 7.46 152.00 ± 9.06 38.50 ± 11.68 B 131 ± 25.86 147.50 ± 22.34 16.50 ± 11.39 C 152.25 ± 15.44 172.75 ± 31.82 20.50 ± 33.41 D 131 ± 12.88 160.00 ± 19.55 29.00 ± 31.35 E 150 ± 4.47 149.75 ± 23.47 -0.75 ± 24.51 F 158.40 ± 20.74 171.00 ± 1.16 13.25 ± 24.73 Values represent Mean ± SD. The body weight analysis demonstrates distinct weight changes among the groups. Group A (control) exhibited a steady increase resulting in a total weight change of 38.50 g. Group B showed a minimal weight gain of only 16.50 g when compared to A, indicating the detrimental effects of alcohol on weight. Group C displayed a weight increase of 20.50 g whilst Groups D, E, and F, were exposed to alcohol and treated with varying doses of ginger. D gained 29.00 g, while Group E had a slight weight loss of 0.75 g, and Group F gained 13.25 g as presented in Table 1 . 3.2 Testicular Morphometry Table 2 Testes Morphometry Groups Testes Weight (g) Testes Size (mm) Gonadosomatic index (%) A 1.57 ± 0.39 2.16 ± 1.76 1.04 ± 0.28 B 1.57 ± 0.32 1.10 ± 0.29 1.10 ± 0.27 C 1.81 ± 0.13 2.61 ± 1.31 1.10 ± 0.31 D 1.75 ± 0.24 1.08 ± 0.28 1.20 ± 0.12 E 1.66 ± 0.28 1.18 ± 0.08 1.11 ± 0.12 F 1.65 ± 0.21 1.31 ± 0.19 0.96 ± 0.13 Values represent Mean ± SD. Testicular morphometric analysis, as shown in Table 2 , reveals that alcohol treatment in Group B did not significantly affect testes weight (1.57 ± 0.32 g) compared to the control Group A (1.57 ± 0.39 g). However, ginger treatment in Group C led to an increase in weight (1.81 ± 0.13 g). Groups D, E, and F, which received ginger after alcohol exposure, showed improved weights (1.75 ± 0.24 g, 1.66 ± 0.28 g, and 1.65 ± 0.21 g, respectively), indicating that ginger mitigated alcohol’s effects. Furthermore, testes size decreased in Group B (1.10 ± 0.29 mm) compared to Group A (2.16 ± 1.76 mm), while Group C saw a size increase (2.61 ± 1.31 mm) with ginger treatment. Groups D, E, and F showed varying increases, all higher than Group B. The gonadosomatic index (GSI) remained similar between Groups A and B but improved slightly in ginger-treated groups, particularly in Group D (1.20 ± 0.12%) while that of Group F had the least signifying the detrimental effects. 3.3 Semen Analysis Table 3 Showing the semen analysis after exposure to alcohol and ginger Groups A B C D E F Motility (%) Progressive motile 71.67 ± 2.89 36.67 ± 2.89 *** 75.00 ± 0.00 51.67 ± 2.89 # 51.67 ± 10.41 # 40 ± 13.23 ### Sluggish motile 23.33 ± 2.89 30.00 ± 5.00 20.00 ± 0.00 26.67 ± 2.89 28.33 ± 2.89 36.67 ± 10.40 # Non-motile 5.00 ± 0.00 33.33 ± 2.89 *** 5.00 ± 0.00 21.67 ± 2.89 20.00 ± 10.00 # 23.33 ± 2.89 Count (10 6 /ml) 37.33 ± 6.43 20.00 ± 7.00 38.33 ± 8.39 34.00 ± 5.00 27.33 ± 11.02 33.00 ± 12.12 Morphology (%) Normal sperm 97.33 ± 0.58 95.33 ± 0.58 95.67 ± 0.58 96.67 ± 1.53 96.00 ± 1.00 95.67 ± 0.58 Head defects Round Head 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 Pinhead 1.00 ± 1.00 3.33 ± 0.58 * 1.67 ± 0.58 0.67 ± 0.58 1.67 ± 1.14 1.33 ± 0.58 # Midpiece defects Bent midpiece 0 0.33 ± 0.58 0 1.00 ± 1.00 1.33 ± 1.15 1.33 ± 0.58 Coiled midpiece 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 Tail defects Headless tail 1.00 ± 0.00 1.33 ± 0.58 1.33 ± 0.58 1.33 ± 0.58 1.00 ± 1.00 0.33 ± 0.58 Coiled tail 0.67 ± 0.58 1.33 ± 0.58 0 0.33 ± 0.58 0 1.33 ± 0.00 Absence of tail 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 Loop tail 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 Values represent Mean ± SD. * represent significance difference when compared to A; # represents significance difference when compared to B. *p < 0.05, **p < 0.001, ***p < 0.0001; #p < 0.05, ##p < 0.001, ###p < 0.0001 As shown in Table 3 , the semen analysis revealed significant differences in motility, count and morphology across the groups. Group B exhibited significantly reduced progressive motile sperms when compared to Group A indicating significant impairment. Group C demonstrated notable improvement when compared to A. Groups D, E and F recorded motility levels significantly different from Group B. Regarding sperm count, Group B had a lower count compared to Group A. Groups C, D, E, and F showed counts higher than Group B. In terms of morphology, the results revealed that Groups A, C, D, and E maintained high percentages of normal sperm (around 95%), while Group B had a slightly lower percentage of 95.33 ± 0.58. Notably, Group B displayed higher instances of head defects, with pinhead defects reported at 3.33 ± 0.58 (p < 0.05) compared to Group A. Additionally, in terms of midpiece defects, Group B had a bent midpiece at 0.33 ± 0.58, while all other groups reported 0.00 ± 0.00. Concerning tail defects, Group B exhibited a higher incidence of headless tails at 1.33 ± 0.58 compared to Group A, which had 1.00 ± 0.00. 3.4 Sperm Chromatin The evaluation of sperm chromatin and DNA integrity, presented in Fig. 1 , revealed significant differences across the experimental groups in a dose dependent manner. In the Aniline Blue staining, Group A had a high percentage of normal chromatin (AB-), recorded at 96.83 ± 1.61, whereas Group B showed a reduction to 92.17 ± 3.62, indicating a detrimental effect on chromatin integrity due to alcohol exposure. Conversely, Groups C, D, E, and F showed high normal chromatin percentages respectively. For the Toluidine Blue staining, which assesses DNA integrity, Group A demonstrated an almost complete absence of abnormal DNA (TB+), with only 0.67 ± 0.76. Group B exhibited a significantly higher percentage of abnormal DNA (TB+) at 12.33 ± 4.04 (p < 0.0001), which is substantially higher than Groups C, D, E, and F, which recorded 0.50 ± 0.50, 4.83 ± 1.89, 2.67 ± 1.26, and 1.00 ± 0.87, respectively. These suggests that Z. officinale treatment effectively mitigates the adverse effects of alcohol on sperm DNA integrity. 3.5 Hormonal Assay As seen in Fig. 2 , Group B exhibited a significant reduction in LH levels compared to Group A indicating alcohol's suppressive effect whereas Group C remained close to control levels. Ginger treatment in groups D, E and F showed mild recovery, with Group D showing the most improvement. Also, alcohol drastically reduced FSH levels in Group B (2.63 mIU/mL, p < 0.0001) compared to Group A (6.57 mIU/mL). Group C remained near the control. Ginger-treated groups D (5.07 mIU/mL, p < 0.05), E (4.30 mIU/mL), and F (4.07 mIU/mL) showed varied degrees of recovery, with Group D achieving better restoration. Nonetheless, alcohol also caused a significant reduction in inhibin B levels in Group B (22.03 pg/mL, p < 0.001) compared to Group A (31.33 pg/mL). Group C (33.00 pg/mL) had elevated levels. Ginger-treated groups D (30.83 pg/mL, p < 0.001), E (27.40 pg/mL), and F (24.67 pg/mL) showed progressive recovery, with Group D showing the greatest restoration. 3.6 Antioxidant Enzyme Level The effects on antioxidant marker is presented in Fig. 3 . GSH levels were significantly decreased in Group B (p < 0.001) compared to Group A. Conversely, Group C had elevated GSH levels at 25.47 U/mg, comparable to Group A. Ginger treatment also improved GSH in Groups D, E and F. 4. Discussion The impact of alcohol and ginger on the body and testicular morphometry in our study showed some variations compared to the existing literature. This can be attributed to differences in experimental design, alcohol dosages, duration of exposure, and animal species used. Obesity is a risk factor for many chronic diseases, such as diabetes, hypertension, and cardiovascular disease [ 28 ]. In our study, alcohol consumption was associated with an increase in body weight, which is consistent with the findings of Kołota et al . [ 29 ], who observed weight gain in Wistar rats after four weeks of 10% ethanol consumption. Epidemiological studies have confirmed that, in adults, excessive alcohol consumption, especially between meals, often leads to metabolic syndrome, with excessive body mass as one of its components [ 30 ]. However, other studies have reported weight loss or stagnation following prolonged alcohol exposure [ 31 , 32 ] primarily due to alcohol’s effects on gastrointestinal function, appetite, and nutrient absorption. The difference between our study and those that report a decrease in body weight is linked to the duration of administration where they administered between 28 days to 12 months. When treated, our study demonstrated a slight increase in body mass in the alcohol-treated rats administered ginger, aligning with findings from Mahamoud & Elnour [ 33 ] and Misawa et al . [ 34 ], where ginger showed potential for reducing adiposity and enhancing lipid metabolism. However, the modest changes in body weight observed in our study are in contrast to other reports where ginger had a more pronounced effect on weight reduction, particularly in high-fat diet models [ 35 ]. This discrepancy could be due that the dose used by these studies was ginger powder and that the later study used women as our study focused on alcohol-treated rats and used aqueous extract, while other studies often explore ginger’s effects on obese or high-fat diet-induced animals, where the metabolic pathways influenced by ginger may be more pronounced. Additionally, the duration of ginger treatment could explain the differences in the design, where more than two (2) weeks were used. For testicular weight, our study found that alcohol led to a decrease in testicular mass, which aligns with studies reporting that alcohol consumption negatively impacts testicular health, often by disrupting spermatogenesis and reducing steroidogenic activity through the decrease in the number of seminiferous tubules and germ cells [ 36 ]. Conversely, ginger administration in our study was associated with increased testicular weight, consistent with studies by Morakinyo et al . [ 37 ] and Khaki et al . [ 38 ], who reported curative effects of ginger on testicular tissue. However, not all studies observed similar outcomes, with Mohammadi et al . [ 18 ] and Akinyemi et al . [ 39 ] findings reporting no significant change in testicular weight with ginger treatment. This variation could be due to differences in the preparation and dosage of ginger used and the timing and method of administration. These studies used combined therapy and pretreated before inducing them with the toxicants. In our study, ginger was administered after alcohol exposure, which may have contributed to its curative effects on testicular weight by counteracting alcohol-induced oxidative stress and inflammation. Studies believe that the increase in testicular weight is related to androgens which are necessary for the development, growth and normal functioning of the testes and male accessory reproductive glands [ 40 ]. Sperm morphology and seminal fluid parameters are considered primary morphological and physicochemical diagnostic markers of male infertility and are crucial for the development of suitable treatments [ 3 ]. With data from animal studies showing that an alcohol-rich diet can affect testicular function, with consequences on the semen quality, the analysis of semen parameters in this study showed significant differences across treatment groups, particularly in sperm motility, count, and morphology, demonstrating the adverse effects of alcohol on reproductive function and the potential curative role of ginger. Previous studies support that alcohol consumption significantly decreases sperm concentration, motility, and morphological integrity [ 41 ]. This reduction is likely due to ethanol's oxidative stress effects, which increase lipid peroxidation in sperm membranes, leading to structural and functional impairments. The marked improvement in motility, count and morphology in ginger-treated groups highlights ginger’s antioxidant properties, consistent with studies by Khaki et al . [ 38 ], Gholami-Ahangaran et al . [ 42 ] which reported that ginger increases sperm parameters and reduces oxidative sperm abnormalities. While the study demonstrates that ginger has a curative role against alcohol-induced reproductive toxicity, the extent of recovery is influenced by the severity and duration of alcohol exposure, as also noted by Biney et al . [ 23 ]. The assessment of DNA damage also offers a tool for selecting sperm with the best DNA integrity for use in assisted reproductive technologies. Chromatin condensation constitutes a valuable parameter in assessing male fertility, completely independent of conventional sperm parameters [ 43 ]. Alcohol consumption is well-documented in the literature as a major factor that compromises sperm DNA integrity through increased DNA damage, reduced chromatin condensation, and heightened rates of germ cell apoptosis [ 44 ]. The impact of alcohol on DNA integrity appears significant, as evidenced by higher abnormal DNA percentages in studies using techniques like TB and AB staining. These effects, observed in several animal models, seem to correlate with the dosage and duration of alcohol exposure, as seen in studies by Rahimipour et al . [ 45 ] and Bai et al . [ 46 ], which report elevated rates of DNA damage and poorly condensed chromatin following ethanol exposure. On the other hand, our study’s exploration of ginger extract as a curative intervention introduces an essential novel aspect, especially given the absence of prior research investigating ginger's effects on sperm chromatin or DNA integrity. Furthermore, our results show that alcohol significantly reduces LH, FSH, and INB levels in the alcohol-untreated group which concurs with studies documenting alcohol’s detrimental effects on the HPG axis [ 47 ]. Interestingly, while our study shows a significant reduction in LH and FSH levels in Group B, some studies have found elevated gonadotropin levels in response to alcohol [ 48 , 49 ] which may be due to the feedback mechanism within the HPG axis. The increase of these hormones in ginger-treated groups resonates with previous findings [ 37 ]. Ginger is known for its antioxidant properties, attributed to bioactive compounds such as gingerols and shogaols. While Basaki et al . [ 50 ] observed decreased GSH activities in alcohol-induced animals, this current study reports an increase in ginger-only and ginger-treated groups as seen in previous studies [ 21 ]. Nonetheless, while ginger is effective in combating alcohol-induced damage, higher doses may not always result in superior benefits and could even lead to diminished responses, a pattern observed in a previous study [ 51 ]. 5. Conclusion This study demonstrated that aqueous Z. officinale extract has a significant curative effect against alcohol-induced reproductive damage in male Wistar rats. Ginger treatment improved testicular morphology and restored semen quality. The extract also boosted antioxidant enzyme activities. Declarations Ethics approval and consent to participate The Research and Ethical Committee of the Faculty of Basic Medical Sciences, with code AE-FUNAI/FBMS/EAHC/24/006, sought ethical approval. Consent for publication Not applicable. Availability of data and materials The datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request. Competing interests The authors declare that they have no competing interests. Funding This study received no funding. Authors' contributions EO & ECN: Contributed to the study design, data analysis, and manuscript writing. NJ, EC & NF: Conducted animal experiments and analysed data. EO, EC & NF: Assisted with semen and biochemical analysis. EO & EC: Reviewed and edited the manuscript. Acknowledgements The authors express their heartfelt gratitude to the Department of Anatomy, Alex Ekwueme Federal University Ndufu-Alike Ikwo (AE-FUNAI), for providing the necessary facilities and support during this research. Special thanks go to the Anatomy and Physiology Laboratory Technologists for their invaluable assistance and technical support, which greatly contributed to the success of this study. References Kyrgiafini M-A, Mamuris Z. Male Infertility: From Genes to Genomes 2022. Genes (Basel). Switzerland; 2023. p. 959. Krausz C, Riera-Escamilla A. Genetics of male infertility. Nat Rev Urol. 2018;15:369–84. Assidi M. Infertility in Men: Advances towards a Comprehensive and Integrative Strategy for Precision Theranostics. Cells. 2022;11:1711. Sun H, Gong T-T, Jiang Y-T, Zhang S, Zhao Y-H, Wu Q-J. 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Effects of Vernonia ambigua on testicular histology, selected semen profiles and serum oxidative stress biomarkers of Wistar rats. J Exp Clin Anat. 2024;21:385–92. Mahomoodally MF, Aumeeruddy MZ, Rengasamy KRR, Roshan S, Hammad S, Pandohee J, et al. Ginger and its active compounds in cancer therapy: From folk uses to nano-therapeutic applications. Semin Cancer Biol. 2021;69:140–9. Kukula-Koch W, Koch W, Czernicka L, Głowniak K, Asakawa Y, Umeyama A, et al. MAO-A Inhibitory Potential of Terpene Constituents from Ginger Rhizomes-A Bioactivity Guided Fractionation. Molecules. 2018;23:1301. Ballester P, Cerdá B, Arcusa R, Marhuenda J, Yamedjeu K, Zafrilla P. Effect of Ginger on Inflammatory Diseases. Molecules. 2022;27:7223. Mohammadi F, Nikzad H, Taghizadeh M, Taherian A, Azami-Tameh A, Hosseini SM, et al. Protective effect of Zingiber officinale extract on rat testis after cyclophosphamide treatment. Andrologia. 2014;46:680–6. Zahedi A, Khaki A. Recovery effect of Zingiber officinale on testis tissue after treatment with gentamicin in rats. J Med Plants Res [Internet]. 2014;8:288–91. Available from: http://academicjournals.org/journal/JMPR/article-abstract/ACCDC2643104 Seif M, Abd El-Aziz T, Sayed M, Wang Z. Zingiber officinale ethanolic extract attenuates oxidative stress, steroidogenic gene expression alterations, and testicular histopathology induced by sodium arsenite in male rats. Environ Sci Pollut Res Int. 2021;28:19783–98. Li N, Xing Y, Sultan AH, Raeeszadeh M, Akbari A, Liu H. Ginger (Zingiber officinale Roscoe) Improves Ethanol-Induced Reproductive Dysfunction by Enhancing Steroidogenesis and Inhibiting Oxidative Stress and Inflammation. Brazilian Arch Biol Technol [Internet]. 2021;64:e21210035. Available from: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132021000100426&tlng=en Otunola GA, Afolayan AJ. Assessment of oral safety profile of aqueous extract blend of three medicinal spices in Wistar rats. Trop J Pharm Res [Internet]. 2017;16:91. Available from: http://www.ajol.info/index.php/tjpr/article/view/150950 Biney RP, Owusu Agyei PE, Ameyaw EO, Afortude JK, Henneh IT, Obese E, et al. Effects of three herb-based alcoholic beverages manufactured in Ghana on sperm characteristics and reproductive hormones in rats. Sci African [Internet]. 2020;7:e00316. Available from: https://linkinghub.elsevier.com/retrieve/pii/S2468227620300545 Ukoha U, Egwu O, Dimkpa U, Maduka S, J.I O, G.U N, et al. Histological and Weight Changes in Testes of Male Albino Rats Fed with Diets Containing Yaji (A Local Meat Sauce). Int J Heal Sci Res. 2014;4:118–26. Famurewa AC, Hamdi H, Sedky A. Lipoic acid abates testis lead accumulation, sperm-endocrine deficits, testicular oxidative inflammation and apoptosis and modulates gene expression of Bax and Bcl-2 in rats. Sci African [Internet]. 2023;21:e01842. Available from: https://www.sciencedirect.com/science/article/pii/S2468227623002983 Ovie FO, Ndukwe GU, Oliver NL, Obi KC, Aguwa US, Olu SI. Effect of Aqueous extract on carica papaya seed and back on the Testes and sperm morphology of male wister rats. Int J Sci Res Publ [Internet]. 2019;9:p9389. Available from: http://www.ijsrp.org/research-paper-0919.php?rp=P939145 Pourmasumi S, Khoradmehr A, Rahiminia T, Sabeti P, Talebi AR, Ghasemzadeh J. Evaluation of Sperm Chromatin Integrity Using Aniline Blue and Toluidine Blue Staining in Infertile and Normozoospermic Men. J Reprod Infertil. 2019;20:95–101. Esomchi CN, Ishicheli GK, Nnaka J, Nwofia JO, Chukwu MA. Prevalence of obesity by occupation among male residents of South East Nigeria. J Anat Sci [Internet]. 2023;14:91–6. Available from: https://www.asn-ng.com/journal/article/1709406443 Kołota A, Głąbska D, Oczkowski M, Gromadzka-Ostrowska J. Influence of Alcohol Consumption on Body Mass Gain and Liver Antioxidant Defense in Adolescent Growing Male Rats. Int J Environ Res Public Health [Internet]. 2019;16:2320. Available from: https://www.mdpi.com/1660-4601/16/13/2320 Vieira BA, Luft VC, Schmidt MI, Chambless LE, Chor D, Barreto SM, et al. Timing and Type of Alcohol Consumption and the Metabolic Syndrome - ELSA-Brasil. PLoS One. 2016;11:e0163044. Ehrlich D, Pirchl M, Humpel C. Effects of long-term moderate ethanol and cholesterol on cognition, cholinergic neurons, inflammation, and vascular impairment in rats. Neuroscience. 2012;205:154–66. Milat AM, Mudnić I, Grković I, Ključević N, Grga M, Jerčić I, et al. Effects of White Wine Consumption on Weight in Rats: Do Polyphenols Matter? Oxid Med Cell Longev. 2017;2017:8315803. Mahmoud RH, Elnour WA. Comparative evaluation of the efficacy of ginger and orlistat on obesity management, pancreatic lipase and liver peroxisomal catalase enzyme in male albino rats. Eur Rev Med Pharmacol Sci. 2013;17:75–83. Misawa K, Hashizume K, Yamamoto M, Minegishi Y, Hase T, Shimotoyodome A. Ginger extract prevents high-fat diet-induced obesity in mice via activation of the peroxisome proliferator-activated receptor δ pathway. J Nutr Biochem. 2015;26:1058–67. Ebrahimzadeh Attari V, Ostadrahimi A, Asghari Jafarabadi M, Mehralizadeh S, Mahluji S. Changes of serum adipocytokines and body weight following Zingiber officinale supplementation in obese women: a RCT. Eur J Nutr. 2016;55:2129–36. Owembabazi E, Nkomozepi P, Mbajiorgu EF. Impact of Concurrent Exposure of Diabetic Male Sprague Dawley Rats to Alcohol and Combination Antiretroviral Therapy (cART) on Reproductive Capacity. Appl Sci [Internet]. 2023;13:5096. Available from: https://www.mdpi.com/2076-3417/13/8/5096 Morakinyo AO, Adeniyi OS, Arikawe AP. Effects of Zingiber Officinale on Reproductive Functions in the Male Rat. African J Biomed Res [Internet]. 2008;11:329–34. Available from: http://www.ajol.info/index.php/ajbr/article/view/50750 Khaki A, Nouri M, Fathiazad F, Khaki A. Evaluation of Zingiber Officinalis and Allium Cepa on Spermatogenesis in Rat. Med J ofTabriz Univ Med Sci Heal Serv [Internet]. 2008;30:53–8. Available from: https://mj.tbzmed.ac.ir/Article/6233 Akinyemi AJ, Adedara IA, Thome GR, Morsch VM, Rovani MT, Mujica LKS, et al. Dietary supplementation of ginger and turmeric improves reproductive function in hypertensive male rats. Toxicol reports. 2015;2:1357–66. Murashima A, Kishigami S, Thomson A, Yamada G. Androgens and mammalian male reproductive tract development. Biochim Biophys Acta - Gene Regul Mech [Internet]. 2015;1849:163–70. Available from: https://www.sciencedirect.com/science/article/pii/S1874939914001266 Oremosu AA, Akang EN. Impact of alcohol on male reproductive hormones, oxidative stress and semen parameters in Sprague–Dawley rats. Middle East Fertil Soc J [Internet]. 2015;20:114–8. Available from: https://www.sciencedirect.com/science/article/pii/S1110569014000661 Gholami-Ahangaran M, Karimi-Dehkordi M, Akbari Javar A, Haj Salehi M, Ostadpoor M. A systematic review on the effect of Ginger (Zingiber officinale) on improvement of biological and fertility indices of sperm in laboratory animals, poultry and humans. Vet Med Sci. 2021;7:1959–69. Kim H-S, Kang MJ, Kim SA, Oh SK, Kim H, Ku S-Y, et al. The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis. Clin Exp Reprod Med. 2013;40:23–8. Silva R, Carrageta DF, Alves MG, Silva BM, Oliveira PF. Antioxidants and Male Infertility. Antioxidants (Basel, Switzerland). Switzerland; 2022. p. 1152. Rahimipour M, Talebi AR, Anvari M, Sarcheshmeh AA, Omidi M. Effects of different doses of ethanol on sperm parameters, chromatin structure and apoptosis in adult mice. Eur J Obstet Gynecol Reprod Biol. 2013;170:423–8. Bai S, Wan Y, Zong L, Li W, Xu X, Zhao Y, et al. Association of Alcohol Intake and Semen Parameters in Men With Primary and Secondary Infertility: A Cross-Sectional Study. Front Physiol. 2020;11:566625. Akbari A, Jelodar GA. The effect of oxidative stress and antioxidants on men fertility. Zahedan J Res Med Sci. 2013;15:1–7. Owembabazi E, Nkomozepi P, Mbajiorgu EF. Androgen Receptor (AR) Depletion Underlies the Reproductive Dysfunctions in Male Rats Exposed to Alcohol and Combination Antiretroviral Therapy (cART). Sarkar D, editor. Andrologia [Internet]. 2023;2023:1–12. Available from: https://www.hindawi.com/journals/and/2023/2966391/ Muthusami KR, Chinnaswamy P. Effect of chronic alcoholism on male fertility hormones and semen quality. Fertil Steril. 2005;84:919–24. Basaki M, Saeb M, Nazifi S, Shamsaei HA. Zinc, copper, iron, and chromium concentrations in young patients with type 2 diabetes mellitus. Biol Trace Elem Res. 2012;148:161–4. Khwanes SA, Mohamed RA, Ibrahim KA, Abd El‐Rahman HA. Ginger reserves testicular spermatogenesis and steroidogenesis in difenoconazole‐intoxicated rats by conducting oxidative stress, apoptosis and proliferation. Andrologia [Internet]. 2022;54:e14241. Available from: https://onlinelibrary.wiley.com/doi/10.1111/and.14241 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5784131","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":401312873,"identity":"8e196c35-2a9a-4dcb-841c-b61b94a7237c","order_by":0,"name":"Collins Nduka Esomchi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0UlEQVRIiWNgGAWjYFCCBGYGhgNAmr0BSBhYkKKFB0QYSJCiRSIBxCNCizl7jrHBjzOH5fgln1/d8KNAgoG/vTsBrxbLnjfGiT03DhtLzs4pu9kDdJjEmbMb8GoxuJFjfIDnw+HEDbdz0m7wALUYSOQS1nLwz4fD9ftvnkm7+YdYLck8Nw4nGEiwH7tNnC1nnhUby5xJN5xxJofttoyBBA9hvxxP3iz55pi1PH/78Wc33/yxkeNv78WvBQnwGIBJYpWDAPsDUlSPglEwCkbBCAIAicBMtJZ0t2EAAAAASUVORK5CYII=","orcid":"","institution":"Alex Ekwueme Federal University Ndufu Alike Ikwo","correspondingAuthor":true,"prefix":"","firstName":"Collins","middleName":"Nduka","lastName":"Esomchi","suffix":""},{"id":401312876,"identity":"aaf5a108-8d99-4ee2-8683-784b287e56cb","order_by":1,"name":"Joy Onyinyechi Nwofia","email":"","orcid":"","institution":"Alex Ekwueme Federal University Ndufu Alike Ikwo","correspondingAuthor":false,"prefix":"","firstName":"Joy","middleName":"Onyinyechi","lastName":"Nwofia","suffix":""},{"id":401312877,"identity":"2d79ed53-76cd-47f1-80b2-8ab45bf9c998","order_by":2,"name":"Favour Kyrian Nwoke","email":"","orcid":"","institution":"Alex Ekwueme Federal University Ndufu Alike Ikwo","correspondingAuthor":false,"prefix":"","firstName":"Favour","middleName":"Kyrian","lastName":"Nwoke","suffix":""},{"id":401312880,"identity":"c4ffa9a5-34b9-4929-98d8-d8be4f3a1f05","order_by":3,"name":"Ogugua Augustine Egwu","email":"","orcid":"","institution":"Alex Ekwueme Federal University Ndufu Alike Ikwo","correspondingAuthor":false,"prefix":"","firstName":"Ogugua","middleName":"Augustine","lastName":"Egwu","suffix":""}],"badges":[],"createdAt":"2025-01-07 20:53:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5784131/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5784131/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":73724693,"identity":"3c974796-f3f1-4404-98f4-0282b4410839","added_by":"auto","created_at":"2025-01-14 03:40:31","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":124121,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eImpact of Alcohol and Ginger on Sperm Chromatin\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eValues represent Mean ± SD. * represent significance difference when compared to A; # represents significance difference when compared to B. *p\u0026lt;0.05, **p\u0026lt;0.001, ***p\u0026lt;0.0001; #p\u0026lt;0.05, ##p\u0026lt;0.001, ###p\u0026lt;0.0001\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5784131/v1/3cf7870f7044e29ab62c3670.png"},{"id":73724960,"identity":"5475d073-27ca-4c4e-87c1-45d8ca29e1de","added_by":"auto","created_at":"2025-01-14 03:48:31","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":97352,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffects of Alcohol and Ginger on Serum Levels of LH, FSH and INB.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eValues represent Mean ± SD. * represent significance difference when compared to A; # represents significance difference when compared to B. *p\u0026lt;0.05, **p\u0026lt;0.001, ***p\u0026lt;0.0001; #p\u0026lt;0.05, ##p\u0026lt;0.001, ###p\u0026lt;0.0001.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5784131/v1/0f65c880b50a4ce044532547.png"},{"id":73724961,"identity":"7aed5422-336f-41c3-864a-05ba2325c439","added_by":"auto","created_at":"2025-01-14 03:48:32","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":49585,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eShowing GSH levels after exposure to alcohol and ginger treatment.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eValues represent Mean ± SD. * represent significance difference when compared to A; # represents significance difference when compared to B. *p\u0026lt;0.05, **p\u0026lt;0.001, ***p\u0026lt;0.0001; #p\u0026lt;0.05, ##p\u0026lt;0.001, ###p\u0026lt;0.0001.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5784131/v1/8c7501a032be6b1248c0aacc.png"},{"id":73725720,"identity":"d42c0634-983f-4ec7-aeba-23d818075469","added_by":"auto","created_at":"2025-01-14 03:56:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1212463,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5784131/v1/7a3cdc91-976d-4e28-a1a2-161e4776f357.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Impact of Zingiber officinale on Testicular Morphometry, Sperm Quality, and Hormonal Profiles in Alcohol-Induced Toxicity","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eInfertility known as the inability to conceive after months of regular, unprotected sexual intercourse for at least 12 months is a significant health matter [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. On different scales, it is known as a medical illness because it affects more than one person instead of a person [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Globally, infertility affects about 15% of couples of reproductive age [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Males account for about 50% of total infertility cases worldwide, the prevalence is said to increase at 0.3% annually leading to an uneven geographical distribution ranging from 20 to 70% [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Nonetheless, the rates of male infertility are underreported due to different factors such as cultural, societal, religious and patriarchal influences that hinder precise sampling and analysis [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Male infertility includes any health condition that hinders the chances of conception. It can result from abnormal sperm function or obstructions preventing ejaculation [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNumerous factors are reported to influence male infertility which include anatomo-pathophysiological factors, environmental factors, the process of ageing and lifestyle factors [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Today, lifestyle-related factors play a significant role in causing male infertility globally [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. An example of these lifestyle-related factors is alcohol consumption which has gained considerable attention due to its global prevalence and modifiable nature [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Alcohol is considered one of the most prevalent dietary factors which people are exposed to and very commonplace in many societies, with almost 60% of the global population aged 15 years and over reported to have consumed alcoholic drinks in a single year [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. The consumption of alcoholic beverages has been a part of the socio-cultural heritage of most populations since ancient times [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Indeed, alcohol has been viewed as an integral part of a meal, and in some cases, as a remedy for infectious diseases or even as a cleaning agent [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Alcohol consumption poses a significant threat to male reproductive health, affecting millions worldwide and impacting testicular structure and function through oxidative stress mechanisms [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Chronic alcohol exposure leads to histological, hormonal, and biochemical alterations, often culminating in reduced semen quality, including impaired sperm concentration and motility\u0026mdash;key markers of male infertility [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSince alcohol causes these distortions via the increase of reactive oxygen species (ROS), many therapeutic approaches focus on synthetic or natural antioxidants derived from natural sources to mitigate the damage caused by an elevation in ROS [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. An example of these natural antioxidants is \u003cem\u003eZingiber officinale\u003c/em\u003e commonly known as ginger. The ginger rhizome is utilised for its aromatic smell and strong taste [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Moreover, ginger is extensively employed in folk medicine for its numerous health benefits in treating various diseases, including chronic conditions such as cancer, diabetes, Alzheimer's, ulcers, cardiovascular disease as well as depression [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The positive impact of ginger on these diseases primarily stems from its antioxidant, antimicrobial, and anti-inflammatory properties [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In different protective studies, ginger has been shown to overcome the reproductive toxicity of cyclophosphamide [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e], gentamicin [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], sodium arsenite [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and ethanol [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] and increase sperm counts, viability, motility, and hormones and improve testicular architecture. Despite these promising findings, the curative potential on testicular morphometry, sperm quality, and hormonal profiles in alcohol-induced toxicity remains underexplored, hence, this study.\u003c/p\u003e"},{"header":"2. Methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Materials\u003c/h2\u003e \u003cp\u003eThe materials for this study include Wistar rats, alcohol solution, \u003cem\u003eZ. officinale\u003c/em\u003e extract, assay kits, laboratory equipment etc.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Study Area\u003c/h2\u003e \u003cp\u003eThis study was carried out in the animal unit of the Anatomy department at Alex Ekwueme Federal University Ndufu Alike Ikwo (AE-FUNAI), Ebonyi State, Nigeria. Ethical approval was sought from the Research and Ethical Committee of the Faculty of Basic Medical Sciences with the code AE-FUNAI/FBMS/EAHC/24/006.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Samples Collection, Identification and Preparations\u003c/h2\u003e \u003cp\u003eFresh samples of ginger tuber were purchased at Ogbe Hausa in Abakaliki Local Government Area, Ebonyi State, Nigeria. The samples were identified in the Applied Biology Department at Ebonyi State University Abakaliki, Nigeria with the voucher number EB/06006. 3g of ginger was shredded into small pieces, dried at room temperature, and then mechanically milled to a fine powder. The obtained solution was dried with a water bath at 40\u0026deg;C, and 2.5g, 5g and 7.5g of the extract were mixed with 30ml, 50ml and 70ml of H\u003csub\u003e2\u003c/sub\u003eO for the low dose, medium dose and high dose treatment, respectively. Otunola \u0026amp; Afolayan [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] report that 5000 mg/kg is not toxic to Wistar rats.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Alcohol Preparation\u003c/h2\u003e \u003cp\u003eThe alcohol (99.7\u0026ndash;100%vv GPR absolute ethanol) produced by NAFCO Scientific Supplies Limited, Surulere Lagos, Nigeria with Product Number 28304 7k was purchased which was used as the toxicant for this experiment. According to the study Biney et al. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], 42% had no mortality in rats.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Animal Care\u003c/h2\u003e \u003cp\u003eThirty (30) male Wistar rats randomly divided into six (6) groups (Groups A-F), each comprising five (5) randomised rats were obtained from the animal house at AE-FUNAI and were kept at room temperature (20\u0026ndash;22\u0026deg;C). The animals were housed in well-ventilated cages with suitable environmental conditions. During their acclimatisation process, which lasted for two weeks at the animal house, the rats were provided with standard feed only.\u003c/p\u003e \u003cp\u003eGroup A served as the normal control group and were administered sterile water throughout the experiment; Group B served as the negative control receiving only 3.50g/kg.bw.d of 40% alcohol for 14 days; Group C served as the positive control and was given a high dose of ginger extract (750 mg/kg.bw) for 14 days; Group D served as an experimental group and was induced with alcohol for 14 days, then treated with a low dose of ginger extracts (250 mg/kg.bw) for another 14 days; Group E as an experimental group was induced with alcohol for 14 days and treated with a medium dose of ginger extracts (500 mg/kg.bw) for 14 days; and, Group F as an experimental group was induced with alcohol for 14 days and treated with a high dose of ginger extracts for 14 days.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Animal Sacrifice and Sample Collection\u003c/h2\u003e \u003cp\u003eOn the 28th day of the experiment, animals were weighed and sacrificed using cervical dislocation. Blood samples were collected by heart puncture and were centrifuged (3000rpm for 15 min) to obtain sera and they were then separated and stored at -80\u0026ordm;C for later hormonal assays. After blood collection, testicular parameters (weight, size and gonadosomatic/testes index) were performed. The left testis was manually homogenized and centrifuged at 3000rpm for 10 min in a cold phosphate buffer (pH 7.4, 0.1M). The obtained supernatant was used to evaluate the level of hormones and antioxidant enzyme activity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Testes Morphometry\u003c/h2\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.7.1 Testis Weight\u003c/h2\u003e \u003cp\u003eThe weight of each testis was recorded immediately after extraction using an electronic scale.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e2.7.2 Testis Size\u003c/h2\u003e \u003cp\u003eA sliding digital Vernier calliper was used to measure the width and length of each testis. The testis size was then calculated using the spheroid formula:\u003c/p\u003e \u003cp\u003eTestis size\u0026thinsp;=\u0026thinsp;width\u003csup\u003e2\u003c/sup\u003e \u0026times; length \u0026times; 0.523 (mm)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e2.7.3 Gonadosomatic/Testes Index\u003c/h2\u003e \u003cp\u003eThe final body and testis weights were used to calculate the gonadosomatic index using the formula previously reported by Ukoha \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:\\text{G}\\text{o}\\text{n}\\text{a}\\text{d}\\text{o}\\text{s}\\text{o}\\text{m}\\text{a}\\text{t}\\text{i}\\text{c}\\:\\text{i}\\text{n}\\text{d}\\text{e}\\text{x}\\:=\\:\\frac{\\text{T}\\text{e}\\text{s}\\text{t}\\text{i}\\text{s}\\:\\text{w}\\text{e}\\text{i}\\text{g}\\text{h}\\text{t}\\:}{\\text{B}\\text{o}\\text{d}\\text{y}\\:\\text{w}\\text{e}\\text{i}\\text{g}\\text{h}\\text{t}\\:}\\:X\\:100\\left(\\%\\right)$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e2.8 Biochemical Assay\u003c/h2\u003e \u003cdiv id=\"Sec14\" class=\"Section3\"\u003e \u003ch2\u003e2.8.1 Hormonal Assay\u003c/h2\u003e \u003cp\u003eThe serum levels of reproductive hormones\u0026mdash;luteinizing hormone (LH), follicle-stimulating hormone (FSH), and inhibin B (INHB)\u0026mdash;were measured using enzyme-linked immunosorbent assay (ELISA) kits. The ELISA kits for LH and FSH were obtained from NIADDK, NIH (USA), while the kit for INHB was sourced from Diagnostic Systems Laboratories (DSL-10-84100i; Webster, TX, USA), following the manufacturer\u0026rsquo;s protocols, as reported by Famurewa \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section3\"\u003e \u003ch2\u003e2.8.2 Glutathione (GSH) Level\u003c/h2\u003e \u003cp\u003eTesticular activities of antioxidant enzyme GSH were analysed in homogenate supernatant using commercial rat ELISA kits.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section3\"\u003e \u003ch2\u003e2.8.3 Semen Analysis\u003c/h2\u003e \u003cp\u003eThe sperm was deposited on a clean, dry glass slide and mixed with an equal volume of 1% NaHCO3 buffered Tyrodes Lactate solution to assess the percentage of sperm motility, count, and morphology, following the procedures outlined by Ovie \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e \u003ch2\u003e2.8.4 Sperm Chromatin Evaluation\u003c/h2\u003e \u003cp\u003eStandard cytochemical methods, incorporating aniline blue (AB) and toluidine blue (TB), were employed to evaluate chromatin condensation and DNA integrity. AB was selectively utilised to stain lysine-rich histones. Air-dried smears obtained from washed semen samples were positioned in 0.2 M phosphate buffer (pH 7.2) containing 3% buffered glutaraldehyde for 30 minutes at room temperature. Subsequently, each smear underwent staining in 4% acetic acid (pH 3.5) with a 5% aqueous solution of AB for 7 minutes. During the light microscopic evaluation, a meticulous count of 200 spermatozoa was conducted across various sections of each slide, utilizing a \u0026times;100 eyepiece magnification [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Sperm heads stained pale blue/colourless and dark blue were considered normal (AB-) and abnormal sperm (AB+), respectively.\u003c/p\u003e \u003cp\u003eOn the other hand, TB served as a metachromatic dye, offering insight into nuclear chromatin condensation and the quality and quantity of DNA fragmentation in sperm. Air-dried sperm smears were fixed using a mixture of 96% ethanol and acetone (1:1) for 30 minutes at a temperature of 4\u0026deg;C. Subsequently, the slides underwent a 5-minute incubation in 0.1 N HCl at 4\u0026deg;C, followed by thorough washing with distilled water three times for 2 minutes each. Finally, staining took place for 10 minutes at room temperature using 0.05% TB in 50% citrate phosphate. In the evaluation process, a minimum of 200 spermatozoa were counted in each sample using light microscopy with a \u0026times;100 eyepiece magnification [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Normal sperm is pale blue and abnormal sperm is dark blue or violet purple. For each sample, the normal (TB-) and abnormal (TB+) spermatozoa were reported as percentages.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e2.9 Data Analysis\u003c/h2\u003e \u003cp\u003eData were subjected to analysis of variance using GraphPad Prism version 8 and presented as Means\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. Group means of parametric data were compared using a one-way analysis of variance, followed by Turkey\u0026rsquo;s post hoc test. p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 Body Weight Analysis\u003c/h2\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eBody weight analysis among the experimental groups.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroups\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eInitial Weight (g)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFinal Weight (g)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWeight change (g)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e113.20\u0026thinsp;\u0026plusmn;\u0026thinsp;7.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e152.00\u0026thinsp;\u0026plusmn;\u0026thinsp;9.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e38.50\u0026thinsp;\u0026plusmn;\u0026thinsp;11.68\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e131\u0026thinsp;\u0026plusmn;\u0026thinsp;25.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e147.50\u0026thinsp;\u0026plusmn;\u0026thinsp;22.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.50\u0026thinsp;\u0026plusmn;\u0026thinsp;11.39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e152.25\u0026thinsp;\u0026plusmn;\u0026thinsp;15.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e172.75\u0026thinsp;\u0026plusmn;\u0026thinsp;31.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20.50\u0026thinsp;\u0026plusmn;\u0026thinsp;33.41\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e131\u0026thinsp;\u0026plusmn;\u0026thinsp;12.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e160.00\u0026thinsp;\u0026plusmn;\u0026thinsp;19.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29.00\u0026thinsp;\u0026plusmn;\u0026thinsp;31.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e150\u0026thinsp;\u0026plusmn;\u0026thinsp;4.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e149.75\u0026thinsp;\u0026plusmn;\u0026thinsp;23.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-0.75\u0026thinsp;\u0026plusmn;\u0026thinsp;24.51\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e158.40\u0026thinsp;\u0026plusmn;\u0026thinsp;20.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e171.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13.25\u0026thinsp;\u0026plusmn;\u0026thinsp;24.73\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eValues represent Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD.\u003c/p\u003e\n \u003cp\u003eThe body weight analysis demonstrates distinct weight changes among the groups. Group A (control) exhibited a steady increase resulting in a total weight change of 38.50 g. Group B showed a minimal weight gain of only 16.50 g when compared to A, indicating the detrimental effects of alcohol on weight. Group C displayed a weight increase of 20.50 g whilst Groups D, E, and F, were exposed to alcohol and treated with varying doses of ginger. D gained 29.00 g, while Group E had a slight weight loss of 0.75 g, and Group F gained 13.25 g as presented in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 Testicular Morphometry\u003c/h2\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eTestes Morphometry\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroups\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTestes Weight\u003c/p\u003e\n \u003cp\u003e(g)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTestes Size\u003c/p\u003e\n \u003cp\u003e(mm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGonadosomatic index (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.61\u0026thinsp;\u0026plusmn;\u0026thinsp;1.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eE\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eValues represent Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD.\u003c/p\u003e\n \u003cp\u003eTesticular morphometric analysis, as shown in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, reveals that alcohol treatment in Group B did not significantly affect testes weight (1.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32 g) compared to the control Group A (1.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39 g). However, ginger treatment in Group C led to an increase in weight (1.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13 g). Groups D, E, and F, which received ginger after alcohol exposure, showed improved weights (1.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24 g, 1.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28 g, and 1.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21 g, respectively), indicating that ginger mitigated alcohol\u0026rsquo;s effects. Furthermore, testes size decreased in Group B (1.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29 mm) compared to Group A (2.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.76 mm), while Group C saw a size increase (2.61\u0026thinsp;\u0026plusmn;\u0026thinsp;1.31 mm) with ginger treatment. Groups D, E, and F showed varying increases, all higher than Group B. The gonadosomatic index (GSI) remained similar between Groups A and B but improved slightly in ginger-treated groups, particularly in Group D (1.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12%) while that of Group F had the least signifying the detrimental effects.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3 Semen Analysis\u003c/h2\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eShowing the semen analysis after exposure to alcohol and ginger\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroups\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eE\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003eMotility (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eProgressive motile\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e71.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51.67\u0026thinsp;\u0026plusmn;\u0026thinsp;10.41\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e40\u0026thinsp;\u0026plusmn;\u0026thinsp;13.23\u003csup\u003e###\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSluggish motile\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23.33\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30.00\u0026thinsp;\u0026plusmn;\u0026thinsp;5.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28.33\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36.67\u0026thinsp;\u0026plusmn;\u0026thinsp;10.40\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNon-motile\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33.33\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.00\u0026thinsp;\u0026plusmn;\u0026thinsp;10.00\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23.33\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003e\u003cstrong\u003eCount (10\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e6\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e/ml)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e37.33\u0026thinsp;\u0026plusmn;\u0026thinsp;6.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.00\u0026thinsp;\u0026plusmn;\u0026thinsp;7.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e38.33\u0026thinsp;\u0026plusmn;\u0026thinsp;8.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34.00\u0026thinsp;\u0026plusmn;\u0026thinsp;5.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e27.33\u0026thinsp;\u0026plusmn;\u0026thinsp;11.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33.00\u0026thinsp;\u0026plusmn;\u0026thinsp;12.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003e\u003cstrong\u003eMorphology (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNormal sperm\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e97.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e96.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e96.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003e\u003cstrong\u003eHead defects\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eRound Head\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePinhead\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003e\u003cstrong\u003eMidpiece defects\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBent midpiece\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCoiled midpiece\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003e\u003cstrong\u003eTail defects\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHeadless tail\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCoiled tail\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbsence of tail\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLoop tail\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eValues represent Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. * represent significance difference when compared to A; # represents significance difference when compared to B. *p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, **p\u0026thinsp;\u0026lt;\u0026thinsp;0.001, ***p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001; #p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, ##p\u0026thinsp;\u0026lt;\u0026thinsp;0.001, ###p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003cp\u003eAs shown in Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, the semen analysis revealed significant differences in motility, count and morphology across the groups. Group B exhibited significantly reduced progressive motile sperms when compared to Group A indicating significant impairment. Group C demonstrated notable improvement when compared to A. Groups D, E and F recorded motility levels significantly different from Group B. Regarding sperm count, Group B had a lower count compared to Group A. Groups C, D, E, and F showed counts higher than Group B. In terms of morphology, the results revealed that Groups A, C, D, and E maintained high percentages of normal sperm (around 95%), while Group B had a slightly lower percentage of 95.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58. Notably, Group B displayed higher instances of head defects, with pinhead defects reported at 3.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) compared to Group A. Additionally, in terms of midpiece defects, Group B had a bent midpiece at 0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58, while all other groups reported 0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00. Concerning tail defects, Group B exhibited a higher incidence of headless tails at 1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 compared to Group A, which had 1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec23\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4 Sperm Chromatin\u003c/h2\u003e\n \u003cp\u003eThe evaluation of sperm chromatin and DNA integrity, presented in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, revealed significant differences across the experimental groups in a dose dependent manner. In the Aniline Blue staining, Group A had a high percentage of normal chromatin (AB-), recorded at 96.83\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61, whereas Group B showed a reduction to 92.17\u0026thinsp;\u0026plusmn;\u0026thinsp;3.62, indicating a detrimental effect on chromatin integrity due to alcohol exposure. Conversely, Groups C, D, E, and F showed high normal chromatin percentages respectively. For the Toluidine Blue staining, which assesses DNA integrity, Group A demonstrated an almost complete absence of abnormal DNA (TB+), with only 0.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76. Group B exhibited a significantly higher percentage of abnormal DNA (TB+) at 12.33\u0026thinsp;\u0026plusmn;\u0026thinsp;4.04 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001), which is substantially higher than Groups C, D, E, and F, which recorded 0.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50, 4.83\u0026thinsp;\u0026plusmn;\u0026thinsp;1.89, 2.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.26, and 1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87, respectively. These suggests that \u003cem\u003eZ. officinale\u003c/em\u003e treatment effectively mitigates the adverse effects of alcohol on sperm DNA integrity.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\n \u003ch2\u003e3.5 Hormonal Assay\u003c/h2\u003e\n \u003cp\u003eAs seen in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, Group B exhibited a significant reduction in LH levels compared to Group A indicating alcohol\u0026apos;s suppressive effect whereas Group C remained close to control levels. Ginger treatment in groups D, E and F showed mild recovery, with Group D showing the most improvement. Also, alcohol drastically reduced FSH levels in Group B (2.63 mIU/mL, p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) compared to Group A (6.57 mIU/mL). Group C remained near the control. Ginger-treated groups D (5.07 mIU/mL, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), E (4.30 mIU/mL), and F (4.07 mIU/mL) showed varied degrees of recovery, with Group D achieving better restoration. Nonetheless, alcohol also caused a significant reduction in inhibin B levels in Group B (22.03 pg/mL, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to Group A (31.33 pg/mL). Group C (33.00 pg/mL) had elevated levels. Ginger-treated groups D (30.83 pg/mL, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), E (27.40 pg/mL), and F (24.67 pg/mL) showed progressive recovery, with Group D showing the greatest restoration.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec25\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6 Antioxidant Enzyme Level\u003c/h2\u003e\n \u003cp\u003eThe effects on antioxidant marker is presented in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. GSH levels were significantly decreased in Group B (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to Group A. Conversely, Group C had elevated GSH levels at 25.47 U/mg, comparable to Group A. Ginger treatment also improved GSH in Groups D, E and F.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe impact of alcohol and ginger on the body and testicular morphometry in our study showed some variations compared to the existing literature. This can be attributed to differences in experimental design, alcohol dosages, duration of exposure, and animal species used. Obesity is a risk factor for many chronic diseases, such as diabetes, hypertension, and cardiovascular disease [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. In our study, alcohol consumption was associated with an increase in body weight, which is consistent with the findings of Kołota \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], who observed weight gain in Wistar rats after four weeks of 10% ethanol consumption. Epidemiological studies have confirmed that, in adults, excessive alcohol consumption, especially between meals, often leads to metabolic syndrome, with excessive body mass as one of its components [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. However, other studies have reported weight loss or stagnation following prolonged alcohol exposure [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] primarily due to alcohol\u0026rsquo;s effects on gastrointestinal function, appetite, and nutrient absorption. The difference between our study and those that report a decrease in body weight is linked to the duration of administration where they administered between 28 days to 12 months. When treated, our study demonstrated a slight increase in body mass in the alcohol-treated rats administered ginger, aligning with findings from Mahamoud \u0026amp; Elnour [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] and Misawa \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e], where ginger showed potential for reducing adiposity and enhancing lipid metabolism. However, the modest changes in body weight observed in our study are in contrast to other reports where ginger had a more pronounced effect on weight reduction, particularly in high-fat diet models [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. This discrepancy could be due that the dose used by these studies was ginger powder and that the later study used women as our study focused on alcohol-treated rats and used aqueous extract, while other studies often explore ginger\u0026rsquo;s effects on obese or high-fat diet-induced animals, where the metabolic pathways influenced by ginger may be more pronounced. Additionally, the duration of ginger treatment could explain the differences in the design, where more than two (2) weeks were used.\u003c/p\u003e \u003cp\u003eFor testicular weight, our study found that alcohol led to a decrease in testicular mass, which aligns with studies reporting that alcohol consumption negatively impacts testicular health, often by disrupting spermatogenesis and reducing steroidogenic activity through the decrease in the number of seminiferous tubules and germ cells [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Conversely, ginger administration in our study was associated with increased testicular weight, consistent with studies by Morakinyo \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] and Khaki \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], who reported curative effects of ginger on testicular tissue. However, not all studies observed similar outcomes, with Mohammadi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] and Akinyemi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] findings reporting no significant change in testicular weight with ginger treatment. This variation could be due to differences in the preparation and dosage of ginger used and the timing and method of administration. These studies used combined therapy and pretreated before inducing them with the toxicants. In our study, ginger was administered after alcohol exposure, which may have contributed to its curative effects on testicular weight by counteracting alcohol-induced oxidative stress and inflammation. Studies believe that the increase in testicular weight is related to androgens which are necessary for the development, growth and normal functioning of the testes and male accessory reproductive glands [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSperm morphology and seminal fluid parameters are considered primary morphological and physicochemical diagnostic markers of male infertility and are crucial for the development of suitable treatments [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. With data from animal studies showing that an alcohol-rich diet can affect testicular function, with consequences on the semen quality, the analysis of semen parameters in this study showed significant differences across treatment groups, particularly in sperm motility, count, and morphology, demonstrating the adverse effects of alcohol on reproductive function and the potential curative role of ginger. Previous studies support that alcohol consumption significantly decreases sperm concentration, motility, and morphological integrity [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. This reduction is likely due to ethanol's oxidative stress effects, which increase lipid peroxidation in sperm membranes, leading to structural and functional impairments. The marked improvement in motility, count and morphology in ginger-treated groups highlights ginger\u0026rsquo;s antioxidant properties, consistent with studies by Khaki \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], Gholami-Ahangaran \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e] which reported that ginger increases sperm parameters and reduces oxidative sperm abnormalities. While the study demonstrates that ginger has a curative role against alcohol-induced reproductive toxicity, the extent of recovery is influenced by the severity and duration of alcohol exposure, as also noted by Biney \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe assessment of DNA damage also offers a tool for selecting sperm with the best DNA integrity for use in assisted reproductive technologies. Chromatin condensation constitutes a valuable parameter in assessing male fertility, completely independent of conventional sperm parameters [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Alcohol consumption is well-documented in the literature as a major factor that compromises sperm DNA integrity through increased DNA damage, reduced chromatin condensation, and heightened rates of germ cell apoptosis [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. The impact of alcohol on DNA integrity appears significant, as evidenced by higher abnormal DNA percentages in studies using techniques like TB and AB staining. These effects, observed in several animal models, seem to correlate with the dosage and duration of alcohol exposure, as seen in studies by Rahimipour \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e] and Bai \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e], which report elevated rates of DNA damage and poorly condensed chromatin following ethanol exposure. On the other hand, our study\u0026rsquo;s exploration of ginger extract as a curative intervention introduces an essential novel aspect, especially given the absence of prior research investigating ginger's effects on sperm chromatin or DNA integrity.\u003c/p\u003e \u003cp\u003eFurthermore, our results show that alcohol significantly reduces LH, FSH, and INB levels in the alcohol-untreated group which concurs with studies documenting alcohol\u0026rsquo;s detrimental effects on the HPG axis [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Interestingly, while our study shows a significant reduction in LH and FSH levels in Group B, some studies have found elevated gonadotropin levels in response to alcohol [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e] which may be due to the feedback mechanism within the HPG axis. The increase of these hormones in ginger-treated groups resonates with previous findings [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Ginger is known for its antioxidant properties, attributed to bioactive compounds such as gingerols and shogaols. While Basaki \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e] observed decreased GSH activities in alcohol-induced animals, this current study reports an increase in ginger-only and ginger-treated groups as seen in previous studies [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Nonetheless, while ginger is effective in combating alcohol-induced damage, higher doses may not always result in superior benefits and could even lead to diminished responses, a pattern observed in a previous study [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e].\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study demonstrated that aqueous \u003cem\u003eZ. officinale\u003c/em\u003e extract has a significant curative effect against alcohol-induced reproductive damage in male Wistar rats. Ginger treatment improved testicular morphology and restored semen quality. The extract also boosted antioxidant enzyme activities.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Research and Ethical Committee of the Faculty of Basic Medical Sciences, with code AE-FUNAI/FBMS/EAHC/24/006, sought ethical approval.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;The authors declare that they have no competing interests.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;This study received no funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEO \u0026amp; ECN: Contributed to the study design, data analysis, and manuscript writing.\u003c/p\u003e\n\u003cp\u003eNJ, EC \u0026amp; NF: Conducted animal experiments and analysed data.\u003c/p\u003e\n\u003cp\u003eEO, EC \u0026amp; NF: Assisted with semen and biochemical analysis.\u003c/p\u003e\n\u003cp\u003eEO \u0026amp; EC: Reviewed and edited the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors express their heartfelt gratitude to the Department of Anatomy, Alex Ekwueme Federal University Ndufu-Alike Ikwo (AE-FUNAI), for providing the necessary facilities and support during this research. Special thanks go to the Anatomy and Physiology Laboratory Technologists for their invaluable assistance and technical support, which greatly contributed to the success of this study.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eKyrgiafini M-A, Mamuris Z. Male Infertility: From Genes to Genomes 2022. Genes (Basel). Switzerland; 2023. p. 959.\u003c/li\u003e\n \u003cli\u003eKrausz C, Riera-Escamilla A. Genetics of male infertility. Nat Rev Urol. 2018;15:369\u0026ndash;84.\u003c/li\u003e\n \u003cli\u003eAssidi M. Infertility in Men: Advances towards a Comprehensive and Integrative Strategy for Precision Theranostics. Cells. 2022;11:1711.\u003c/li\u003e\n \u003cli\u003eSun H, Gong T-T, Jiang Y-T, Zhang S, Zhao Y-H, Wu Q-J. Global, regional, and national prevalence and disability-adjusted life-years for infertility in 195 countries and territories, 1990-2017: results from a global burden of disease study, 2017. 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Available from: https://onlinelibrary.wiley.com/doi/10.1111/and.14241\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Alcohol-induced toxicity, Antioxidant, Ginger, Semen quality, Testicular morphometry Zingiber officinale","lastPublishedDoi":"10.21203/rs.3.rs-5784131/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5784131/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eAlcohol consumption is known to induce reproductive toxicity, leading to adverse effects on testicular morphology, sperm quality, and DNA integrity in males. \u003cem\u003eZingiber officinale \u003c/em\u003e(ginger), known for its antioxidant and anti-inflammatory properties, may counteract these effects. This study aimed to investigate the impact of ginger on testicular morphometry, sperm quality, and hormonal profiles in alcohol-induced toxicity. 30 male Wistar rats were divided into six groups (n=5/group). Group A (control) received normal saline. Group B was exposed to 40% alcohol (3.50 g/kg body weight) from Days 15–28. Group C received ginger (750 mg/kg) during the same period. Groups D, E, and F were treated with alcohol for 14 days, followed by low (250 mg/kg), medium (500 mg/kg), and high (750 mg/kg) doses of ginger, respectively, from Days 15–28. The study evaluated changes in body and testicular morphometry, antioxidant enzyme and hormonal changes. Semen analysis included sperm motility, count, and morphology, while sperm chromatin/DNA integrity was assessed using Aniline Blue and Toluidine Blue staining.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eAlcohol exposure (Group B) significantly reduced testicular weight, sperm motility, and chromatin integrity. Ginger-treated groups (C, D, and E) showed marked improvements, with Group C outperforming the control group (A) in sperm motility, antioxidant levels, and hormones. However, Group F showed distorted results similar to Group B, suggesting high-dose toxicity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: Ginger improves reproductive health and mitigates alcohol-induced toxicity in a dose-dependent manner. Moderate doses show optimal benefits, while high doses may be detrimental. These findings support ginger’s potential as a natural therapeutic agent for reproductive health.\u003c/p\u003e","manuscriptTitle":"Impact of Zingiber officinale on Testicular Morphometry, Sperm Quality, and Hormonal Profiles in Alcohol-Induced Toxicity","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-14 03:40:27","doi":"10.21203/rs.3.rs-5784131/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b30d6011-5de1-4148-bd25-f29c4767b5ab","owner":[],"postedDate":"January 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-01-14T03:40:27+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-14 03:40:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5784131","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5784131","identity":"rs-5784131","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}