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    <title>Oral administration of Fumaria officinalis alleviates nephrotoxicity and improves behavior in Japanese quails</title>
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    <!-- Article Title -->
    <h1>Oral administration of <i>Fumaria officinalis</i> alleviates nephrotoxicity and improves behavior in Japanese quails</h1>

    <!-- Authors -->
    <div class="authors">
        Maryam Saleem<sup>1</sup>, Saba Malik<sup>2*</sup>, Iftikhar Hussain<sup>3</sup>, Rohina Tabassam<sup>1</sup>, Hafiza Sadia<sup>2</sup>, Sheeba Riaz<sup>1</sup>, Namrah<sup>1</sup>, Muhammad Sakandar Majid<sup>1*</sup>
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    <!-- Affiliations -->
    <div class="affiliations">
        <sup>1</sup>Department of Zoology, Wildlife & Fisheries, University of Agriculture, Faisalabad, Pakistan.<br>
        <sup>2</sup>State Key Laboratory of Water Pollution Control and Green Resource Recycling, School of the Environment, Nanjing University, Nanjing, Jiangsu 210023, China.<br>
        <sup>3</sup>State Key Laboratory of Pharmaceutical Biotechnology, NJU Advanced Institute for Life Sciences (NAILS), School of Life Sciences, Nanjing University, Nanjing 210023, China.
    </div>

    <!-- Article Meta Info -->
    <div class="article-meta">
        <strong>Article Info</strong><br>
        <strong>Academic Editor:</strong> Muhammad Naeem<br>
        <strong>Received:</strong> 21 January 2026 &nbsp;|&nbsp; <strong>Accepted:</strong> 28 February 2026 &nbsp;|&nbsp; <strong>Published:</strong> 5 March 2026<br>
        <strong>Citation:</strong> Saleem, M., Malik, S., Hussain, I., Tabassam, R., Sadia, H., Riaz, S., Namrah, & Majid, M. S. (2026). Oral administration of <i>Fumaria officinalis</i> alleviates nephrotoxicity and improves behavior in Japanese quails. <em>Pakistan Journal of Zoological Sciences</em>, <strong>2</strong>(1), 1-10.
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    <!-- DOI -->
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        <strong>DOI:</strong> <a href="https://doi.org/10.65139/76ddxr40" target="_blank">https://doi.org/10.65139/76ddxr40</a>
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    <!-- Abstract -->
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        <strong>Abstract</strong>
        <p>Aromatic hydrocarbons (AHCs) are crucial in petroleum-based transportation and consumer product manufacturing, but their accumulation in the body poses serious threats. Toluene, an important aromatic hydrocarbon, can cause oxidative stress and structural damage in the liver, kidney, and brain due to its lipophilic nature. <em>Fumaria officinalis</em>, an antioxidant with flavonoids and alkaloids, has been shown to protect against renal diseases. The study aims to demonstrate the beneficial effects of <em>F. officinalis</em> on general performance and in reducing nephrotoxicity caused by toluene in Japanese quail. After a period of acclimation, sixteen Japanese quails, weighing between 100 and 140g, were used in the experiment. Four equal groups of birds were created: one group was given tap water and standard food; another group was exposed to toluene (0.2 mL/kg) in distilled water; a combination treatment of toluene (0.2 mL/kg) and <em>F. officinalis</em> (150 mg/kg B.W.) was given to the third group; and <em>F. officinalis</em> (150 mg/kg B.W.) was given to the fourth group. After a month, the birds were slaughtered, and blood samples were collected for analysis. The toluene-exposed group showed a significant increase in uric acid, creatinine, and biliverdin levels, a decline in albumin levels, and a decrease in body weight gain. The enzyme activity of superoxide dismutase and catalase was significantly lower in the toluene group. Furthermore, <em>F. officinalis</em> improved toluene-induced reductions in alertness, feeding activity, crowing, mating activity, and overall behavioral performance. This suggests that <em>F. officinalis</em> protects quails' kidneys against the toxicity of toluene.</p>
    </div>

    <!-- Keywords -->
    <div class="keywords">
        <strong>Keywords:</strong> Toluene; Nephrotoxicity; Oxidative Stress; Antioxidant Enzymes; Renal Protection
    </div>

    <hr>

    <!-- Introduction -->
    <h2>1. Introduction</h2>
    <p>Environmental toxins are a major concern in modern toxicology because many industrial, agricultural, and transportation-related chemicals persist in air, soil, water, and biological systems [9]. Among these pollutants, aromatic hydrocarbons are considered important environmental contaminants due to their widespread use and potential to affect multiple organs after exposure [9; 5]. Monocyclic aromatic...</p>
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    <!-- Materials and Methods -->
    <h2>2. Materials and Methods</h2>
    <h3>2.1 Experimental Animals and Ethical Statement</h3>
    <p>Sixteen healthy quails, aged 4-6 weeks and weighing 110-140 g, were obtained from a nearby hatchery. The birds were acclimatized for seven days under controlled laboratory conditions before the start of the experiment. During acclimatization, the quails were maintained at 25±2°C with 50-60% relative humidity and a 12h light / 12h dark photoperiod. All birds received commercial feed chow and fresh drinking water ad libitum. The Departmental Animal Ethics Committee, University of Agriculture, Faisalabad, approved the experimental protocol. All procedures were performed according to institutional guidelines for the care and use of experimental animals.</p>

    <h3>2.2 Experimental Design</h3>
    <p>After acclimatization, the quails were divided into four equal experimental groups, with four birds in each group (n=4). The experimental period lasted for 30 days. The control group received normal feed and tap water under a clean environment at room temperature [33]. The toluene-treated group received toluene at a dose of 0.2 mL/kg body weight through oral gavage. The co-treated group received both toluene at 0.2 mL/kg body weight and <em>Fumaria officinalis</em> extract at 150 mg/kg body weight through oral gavage. The <em>F. officinalis</em> control group received only <em>F. officinalis</em> extract at 150 mg/kg body weight through oral gavage. All treatments were administered once daily for 30 consecutive days using a sterile feeding needle.</p>

    <!-- Table 1: Experimental groups -->
    <h3>Table 1</h3>
    <p><strong>Experimental groups and treatments</strong></p>
    <table>
        <thead>
            <tr>
                <th>Group</th>
                <th>Treatment</th>
                <th>Dose</th>
                <th>Route</th>
                <th>Duration</th>
            </tr>
        </thead>
        <tbody>
            <tr>
                <td>Group I</td>
                <td>Normal control</td>
                <td>Feed and tap water only</td>
                <td>Oral intake</td>
                <td>30 days</td>
            </tr>
            <tr>
                <td>Group II</td>
                <td>Toluene-treated group</td>
                <td>Toluene, 0.2 mL/kg body weight</td>
                <td>Oral gavage</td>
                <td>30 days</td>
            </tr>
            <tr>
                <td>Group III</td>
                <td>Co-treated group</td>
                <td>Toluene, 0.2 mL/kg + <em>F. officinalis</em>, 150 mg/kg</td>
                <td>Oral gavage</td>
                <td>30 days</td>
            </tr>
            <tr>
                <td>Group IV</td>
                <td><em>F. officinalis</em> control group</td>
                <td><em>F. officinalis</em>, 150 mg/kg body weight</td>
                <td>Oral gavage</td>
                <td>30 days</td>
            </tr>
        </tbody>
    </table>

    <!-- Continue with other sections like Results, Discussion, References -->
    <h2>3. Results</h2>
    <h3>3.1 Body and Kidney Weight Alterations</h3>
    <p>There were notable differences (p < 0.05) in body weight between the groups. Quails exposed to toluene exhibited a significant decrease in body weight (31.25±9.47g) in contrast to the control group (46.5±1.85g). Weight was partially restored (42.25±8.15 g) by co-treatment with <em>Fumaria officinalis</em>, indicating a statistical improvement over the toluene group but not a significant difference from the control. Only <em>F. officinalis</em>-treated quails showed a non-significant increase in weight (33±8.36g).</p>

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    <div class="references">
        <h2>References</h2>
        <p>[1] Abdel-Salam, O. M. E., Sleem, A. A., & Shaffie, N. M. (2020). Antioxidant and antiinflammatory effects of toluene exposure in rats. <em>Environmental Science and Pollution Research</em>, 27(5), 5151-5161. DOI: 10.21608/ejchem.2022.135813.5983.</p>
        <p>[2] Abou-Zeid, S. M., Abdel-Rahman, M. A., & Sharaf, H. A. (2018). Effect of toluene inhalation on antioxidant status and the protective role of quercetin in rats. <em>Biochemistry and Cell Biology</em>, 96(6), 676-684. doi: 10.1080/09674845.2008.11732801.</p>
        <p>[3] Abduljalel, M. E., & Al-Saadi, R. N. (2022). Toxicopathological effect of benzene, toluene, ethylbenzene, and xylenes (BTEX) as a mixture and the protective effect of citicoline in male rats following 90-day oral exposure. <em>REDVET- Revista electrónica de Veterinaria</em>, 23(3).</p>
        <p>[4] Tienda-Vázquez, M. A., Morreeuw, Z. P., Sosa-Hernández, J. E., Cardador-Martínez, A., Sabath, E., Melchor-Martínez, E. M., ... & Parra-Saldivar, R. (2022). Nephroprotective plants: a review on the use in pre-renal and post-renal diseases. <em>Plants</em>, 11(6), 818. https://doi.org/10.3390/plants11060818</p>
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        <strong>Copyright:</strong> © 2026 by the authors. This is an open access article distributed under the Creative Commons Attribution (CC BY) license. © 2026 Published by IJSMART Publishing Company.<br><br>
        <strong>Disclaimer:</strong> Pakistan Journal of Zoological Sciences (PJZS) is an open-access journal. All articles published in the journal are distributed under the terms of the Creative Commons Attribution-NonCommercial License (CC BY-NC). This permits use, distribution, and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. The statements, opinions, and data presented in the publications of PJZS are solely those of the individual authors and contributors. They do not necessarily reflect the views of IJSmart Publishing Company or the journal's editors. IJSmart Publishing Company and the editors disclaim any liability for any injury, loss, or damage to persons or property resulting from the use of ideas, methods, instructions, or products mentioned in the content.
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