Assessing Compliance with ARRIVE Guidelines in Zebrafish Herbal Toxicity Studies: Are We Meeting the Standards?
DOI:
https://doi.org/10.48165/jlas.2026.9.2.5Keywords:
ARRIVE guidelines, Zebrafish, Danio rerio, Embryotoxicity, Herbal toxicityAbstract
Transparent reporting of animal studies is essential for scientific reproducibility, whereas inadequate reporting hinders the replication of experimental methods. The ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines were developed to standardize the reporting of animal research. This study evaluated the reporting quality of zebrafish embryotoxicity studies investigating herbal medicines, using the ARRIVE-2020 guidelines as a reference. Original research articles published between 2020 and 2023 were assessed for compliance with the 10 Essential ARRIVE items. Eight studies from eight different journals were included, and both journal and article-related characteristics were examined. Each ARRIVE item was categorized as “fully reported”, “partially reported”, or “not reported”, and overall com pliance was classified as excellent (≥80%), moderate (50-79%), or poor (≤49%) based on the proportion of fully reported items. Among the analysed journals, 87.5% were indexed in SCI-Expanded, and 75% endorsed ARRIVE guidelines in their author instructions. The proportion of fully reported items ranged from 0% to 100%, while partially reported items ranged from 0% to 37.5%. Compliance was highest for study design, sample size, outcome measures, experimental animals, and reported outcomes. In contrast, inclusion and exclusion criteria, as well as randomization, were consistently poorly reported. These findings reveal substantial deficiencies in the reporting of key ARRIVE items in zebrafish embryotoxicity research. Although ARRIVE guidelines have the potential to improve reporting quality, their limited implementation may reflect insufficient author compliance and inadequate journal enforcement. We encourage the full and consistent application of ARRIVE guidelines to enhance research quality, reproducibility, and zebrafish welfare.
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References
Liu, Y., Yang, G., Yang, C., Shi, Z., Ru, Y., Shen, N., et al. (2023). The mechanism of Houttuynia cordata embryotoxicity was explored in combination with an experimental model and network pharmacology. Toxins, 15(1), 73. https://doi.org/10.3390/toxins15010073
Kilkenny, C., Browne, W. J., Cuthill, I. C., Emerson, M., & Altman, D. G. (2010). Improving bioscience research reporting: The ARRIVE guidelines for reporting animal research. PLoS Biology, 8(6), e1000412. https://doi.org/10.1371/journal.pbio.1000412
Hooijmans, C. R., Leenaars, M., & Ritskes-Hoitinga, M. (2010). Gold standard publication checklist to improve the quality of animal studies, to fully integrate the Three Rs, and to make systematic reviews more feasible. Alternatives to Laboratory Animals, 38(2), 167–182. https://doi.org/10.1177/026119291003800208
Song, J., Solmi, M., Carvalho, A., Shin, J. I., & Ioannidis, J. P. A. (2023). Twelve years after the ARRIVE guidelines: Animal research has not yet arrived at high standards. Laboratory Animals, 57(6), 507–513. https://doi.org/10.1177/00236772231172116
Rodwell, V., Patil, M., Kuht, H. J., Stephan, C. F. N., William, H. J. N., & Mervyn, G. T. (2024). Zebrafish optokinetic reflex: Minimal reporting guidelines and recommendations. Biology, 13(4), 250. https://doi.org/10.3390/biology13040250
Sert, N. P. du, Ahluwalia, A., Alam, S., Avey, M. T., Baker, M., Browne, W. J., et al. (2020). Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0. PLoS Biology, 18(7), e3000411. https://doi.org/10.1371/journal.pbio.3000411
Cassar, S., Adatto, I., Freeman, J. L., Gamse, J. T., Iturria, I., Lawrence, C., et al. (2020). Use of zebrafish in drug discovery toxicology. Chemical Research in Toxicology, 33(1), 95–118. https://doi.org/10.1021/acs.chemrestox.9b00335
Schroeder, P. G. (2022). The welfare of zebrafish. In L. D'Angelo & P. de Girolamo (Eds.), Laboratory fish in biomedical research (pp. 101–117). Academic Press.
National Centre for the Replacement, Refinement and Reduction of Animals in Research. (2020). The ARRIVE Essential 10: Compliance questionnaire. https://arriveguidelines.org/resources/compliance-questionnaire
Syed, M. S. N. A., Khatib, A., So’ad, S. Z. M., Ahmed, Q. U., Ibrahim, Z., Nipun, T. S., et al. (2023). In vitro anti-diabetic, anti-inflammatory, antioxidant activities and toxicological study of optimized Psychotria malayana Jack leaves extract. Pharmaceuticals, 16, 1692. https://doi.org/10.3390/ph16121692
Nguyen, T. H., Nguyen, P. D., Quetin-Leclercq, J., Muller, M., Ly Huong, D. T., Pham, H. T., et al. (2020). Developmental toxicity of Clerodendrum cyrtophyllum Turcz. ethanol extract in zebrafish embryo. Journal of Ethnopharmacology, 249, 112384. https://doi.org/10.1016/j.jep.2019.112384
Veeren, B., Ghaddar, B., Bringart, M., Khazaal, S., Gonthier, M.-P., Meilhac, O., et al. (2020). Phenolic profile of herbal infusion and polyphenol-rich extract from leaves of the medicinal plant Antirhea borbonica: Toxicity assay determination in zebrafish embryos and larvae. Molecules, 25(19), 4482. https://doi.org/10.3390/molecules25194482
Gu, L., Wang, X., Shao, X., Ding, Y., & Li, Y. (2022). Study on chemical constituents of Folium Artemisiae argyi Carbonisatum, toxicity evaluation on zebrafish, and intestinal hemostasis. Saudi Pharmaceutical Journal, 30(5), 532–543. https://doi.org/10.1016/j.jsps.2022.02.011
Ahda, M., Jaswir, I., Khatib, A., Ahmed, Q. U., Mahfudh, N., Ardini, Y. D., et al. (2023). Phytochemical analysis, antioxidant, α-glucosidase inhibitory activity, and toxicity evaluation of Orthosiphon stamineus leaf extract. Scientific Reports, 13(1), 17012. https://doi.org/10.1038/s41598-023-44176-4
Farooq, M., Abutaha, N., Mahboob, S., Baabbad, A., Almoutiri, N. D., & Wadaan, M. A. A. M. (2020). Investigating the antiangiogenic potential of Rumex vesicarius (humeidh), anticancer activity in cancer cell lines, and assessment of developmental toxicity in zebrafish embryos. Saudi Journal of Biological Sciences, 27(2), 611–622. https://doi.org/10.1016/j.sjbs.2019.11.003
Xia, Z., Hao, E., Chen, Z., Zhang, M., Wei, Y., Wei, M., et al. (2020). Roots and stems of Kadsura coccinea extract induced developmental toxicity in zebrafish embryos/larvae through apoptosis and oxidative stress. Pharmaceutical Biology, 58(1), 1294–1301. https://doi.org/10.1080/13880209.2020.1857737
Barros, T. P., Alderton, W. K., Reynolds, H. M., Roach, A. G., & Berghmans, S. (2008). Zebrafish: An emerging technology for in vivo pharmacological assessment to identify potential safety liabilities in early drug discovery. British Journal of Pharmacology, 154(7), 1400–1413. https://doi.org/10.1038/bjp.2008.249
Kimmel, C. B., Ballard, W. W., Kimmel, S. R., Ullmann, B., & Schilling, T. F. (1995). Stages of embryonic development of the zebrafish. Developmental Dynamics, 203(3), 253–310. https://doi.org/10.1002/aja.1002030302
Jalgaonkar, S., Mapara, T., Verma, A., & Sayyed, M. (2019). Comparison of adherence to ARRIVE guidelines in animal research articles published in the years 2009 and 2016 in pharmacology journals: An observational study. Journal of Pharmacology & Pharmacotherapeutics, 10(2), 77–84. https://doi.org/10.4103/jpp.JPP_104_18
Gulin, J. E. N., Rocco, D. M., & García-Bournissen, F. (2015). Quality of reporting and adherence to ARRIVE guidelines in animal studies for Chagas disease preclinical drug research: A systematic review. PLoS Neglected Tropical Diseases, 9(11), e0004194. https://doi.org/10.1371/journal.pntd.0004194
Baker, D., Lidster, K., Sottomayor, A., & Amor, S. (2014). Two years later: Journals are not yet enforcing the ARRIVE guidelines on reporting standards for pre-clinical animal studies. PLoS Biology, 12(1), e1001756. https://doi.org/10.1371/journal.pbio.1001756
Alemán-Laporte, J., Alvarado, G., García-Gómez, M. S., Fonseca, B. A. A. T., Zúñiga-Montero, M., & Cabrera, M. C. M. (2019). Quality of adherence to the ARRIVE guidelines in the materials and methods section in studies where swine were used as surgical biomodels: A systematic review. Animals, 9(11), 947. https://doi.org/10.3390/ani9110947
