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Fisheries Science of Ukraine, 2026; 3(77): 152-171
DOI: https://doi.org/10.61976/fsu2026.03.152
UDC 577.121:594.381.5:574.64

Received: 30.07.2026
Received in revised form: 28.08.026
Published: 30.09.2026

Dynamics of β-carotene content in the hemolymph and hepatopancreas of Lymnaea stagnalis (Linnaeus, 1758) under chronic phenol exposure

L. Muzyka, This email address is being protected from spambots. You need JavaScript enabled to view it. , ORCID ID 0000-0001-7752-7853, Zhytomyr Ivan Franko State University, Zhytomyr
G. Kyrychuk, This email address is being protected from spambots. You need JavaScript enabled to view it. , ORCID ID 0000-0002-1059-2834, Zhytomyr Ivan Franko State University, Zhytomyr

Purpose. To establish the patterns of change in the β-carotene content in the hemolymph and hepatopancreas of Lymnaea stagnalis under the effect of phenol at concentrations of 0.0005, 0.002, 0.005 and 0.01 mg/dm3 at different exposure durations (2, 7, 14, and 21 days).

Methodology. The study was conducted on sexually mature, uniformly sized L. stagnalis (Linnaeus, 1758) mollusks. The animals were kept in phenol solutions at concentrations of 0.0005, 0.002, 0.005 and 0.01 mg/dm3 for 2, 7, 14, and 21 days. All experiments were accompanied by a control. The β-carotene content was determined spectrophotometrically in the hemolymph and in previously homogenized hepatopancreas tissues after extraction with hexane (1:4). Two-way analysis of variance (two-way ANOVA) and Tukey’s post hoc test (Tukey HSD) were used for the statistical processing of the results.

Findings. Two-way analysis of variance showed a statistically significant effect of the factors “phenol concentration” and “exposure duration” as well as of their interaction on the β-carotene content in the hepatopancreas (p < 0.001) and hemolymph (p < 0.001) of the mollusks studied, which indicates the nonlinear and time-dependent nature of the response of the carotenoid system. In the hepatopancreas of L. stagnalis, at the early stage of exposure (2 days) under the effect of phenol at a concentration of 0.0005 and 0.005 mg/dm3, the β-carotene content decreased significantly, by 42.4–46.8% (p < 0.001–0.01), whereas prolongation of the exposure to 21 days was accompanied by an increase in this parameter by 31.2–51.5 % (p < 0.01–0.05), which indicates a reorganization of carotenoid metabolism under conditions of prolonged toxic effect. In the hemolymph of the mollusks studied, the changes were more variable; however, on day 21 of exposure, the effect of phenol, irrespective of the concentration, caused an increase in the β-carotene content by 15.83–46.95 % relative to the control (p < 0.01–0.05).

Originality. The temporal and tissue-specific dynamics of β-carotene content in the hemolymph and hepatopancreas of Lymnaea stagnalis under chronic phenol exposure within the concentration range of 0.0005–0.01 mg/dm3 were established, and the pattern of these changes was shown to depend on the combined effects of phenol concentration and exposure duration.

Practical Value. The results obtained expand current understanding of the reorganization of carotenoid metabolism in freshwater mollusks under conditions of phenol intoxication and substantiate the potential of using β-carotene as a biochemical marker of phenol pollution in the ecotoxicological monitoring of freshwater ecosystems.

Keywords: freshwater mollusks, aquatic bioresources, aquatic toxicology, carotenoids, ecotoxicological monitoring.

REFERENCES

  1. Ekelund Ugge, G. M., Jonsson, A., & Berglund, O. (2022). Molecular biomarker responses in the freshwater mussel Anodonta anatina exposed to an industrial wastewater effluent. Environmental Science and Pollution Research, 29(2), 2158–2170. https://doi.org/10.1007/s11356-021-15633-4
  2. Atkinson, C. L., Hopper, G. W., Kreeger, D. A., Lopez, J. W., Maine, A. N., Sansom, B. J., Schwalb, A., & Vaughn, C. C. (2023). Gains and gaps in knowledge surrounding freshwater mollusk ecosystem services. Freshwater Mollusk Biology and Conservation, 26(1), 20–31. https://doi.org/10.31931/fmbc-d-22-00002 
  3. Strong, E. E., Gargominy, O., Ponder, W. F., & Bouchet, P. (2008). Global diversity of gastropods (Gastropoda; Mollusca) in freshwater. Hydrobiologia, 595(1), 149–166. https://doi.org/10.1007/s10750-007-9012-6
  4. Panda, F., Pati, S. G., Das, K., Samanta, L., Sahoo, D. K., & Paital, B. (2022). Biochemical and molecular responses of the freshwater snail Pila sp. to environmental pollutants, abiotic, and biotic stressors. Frontiers in Environmental Science, 10, 1033049. https://doi.org/10.3389/fenvs.2022.1033049
  5. Al-Alam, J., Millet, M., Khoury, D., Rodrigues, A., Akoury, E., Tokajian, S., & Wazne, M. (2024). Biomonitoring of PAHs and PCBs in industrial, suburban, and rural areas using snails as sentinel organisms. Environmental Science and Pollution Research, 31(3), 4970–4984. https://doi.org/10.1007/s11356-023-31493-6
  6. Kutluyer Kocabaş, F., Gцktьrk Aksu, E., & Kocabaş, M. (2023). Evaluation of metal pollution related to human health risk in freshwater snail Viviparus contectus (Millet, 1813) as a potential bioindicator species in Lake Habitat (Turkey). Environmental Science and Pollution Research, 30(41), 93686–93696. https://doi.org/10.1007/s11356-023-29062-y
  7. Voloshyna, O. M., & Krupiei, K. S. (2015). Vplyv fenolu ta formaldehidu na pihmentosyntezuvalnu zdatnist vodnykh mikroorhanizmiv [Influence of phenol and formaldehyde on the pigment-synthesizing ability of the microorganisms]. Pytannia bioindykatsii ta ekolohii [Problems of bioindications and ecology], 20(1), 166–179.
  8. Guerra, R. (2001). Ecotoxicological and chemical evaluation of phenolic compounds in industrial effluents. Chemosphere, 44(8), 1737–1747. https://doi.org/10.1016/S0045-6535(00)00562-2
  9. Ramos, R. L., Moreira, V. R., & Amaral, M. C. S. (2024). Phenolic compounds in water: Review of occurrence, risk, and retention by membrane technology. Journal of Environmental Management, 351, 119772. https://doi.org/10.1016/j.jenvman.2023.119772
  10. Anku, W. W., Mamo, M. A., & Govender, P. P. (2017). Phenolic compounds in water: sources, reactivity, toxicity and treatment methods. Phenolic compounds – natural sources, importance and applications, 419–443. https://doi.org/10.5772/66927
  11. Zeng, X., Yu, J., Zhang, S., Ni, T., & Ma, D. (2023). Ecological risk of phenol on typical biota of the northern Chinese river from an integrated probability perspective: The Hun River as an example. Environmental Monitoring and Assessment, 195(12), 1512. https://doi.org/10.1007/s10661-023-12089-6
  12. Muthukumaravel, K., Priyadharshini, M., Kanagavalli, V., Vasanthi, N., Ahmed, M. S., Musthafa, M. S., ... & Ravindran, B. (2023). Impact of sublethal phenol in freshwater fish Labeo rohita on biochemical and haematological parameters. Environmental Monitoring and Assessment, 195(1), 10. https://doi.org/10.1007/s10661-022-10554-2
  13. Muthukumaravel, K., Kanagavalli, V., Pradhoshini, K. P., Vasanthi, N., Santhanabharathi, B., Alam, L., Musthafa, M. S., & Faggio, C. (2023). Potential biomarker of phenol toxicity in freshwater fish C. mrigala: Serum cortisol, enzyme acetylcholine esterase and survival organ gill. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 263, 109492. https://doi.org/10.1016/j.cbpc.2022.109492
  14. Chakraborty, D., Mandal, A. H., Ghosh, S., Sadhu, A., Das, D., Saha, N. C., & Saha, S. (2024). The physiological effects of acute and sub-lethal exposure to phenol on antioxidant enzyme activity in the freshwater sludge worm Tubifex tubifex. Toxicology Reports, 13, 101717. https://doi.org/10.1016/j.toxrep.2024.101717
  15. Gami, A. A., Shukor, M. Y., Khalil, K. A., Dahalan, F. A., Khalid, A., & Ahmad, S. A. (2014). Phenol and its toxicity. Journal of Environmental Microbiology and Toxicology, 2(1), 11–23. https://doi.org/10.54987/jemat.v2i1.89
  16. Lemos, M. F. (2021). Biomarker studies in stress biology: from the gene to population, from the organism to the application. Biology, 10(12), 1340. https://doi.org/10.3390/biology10121340
  17. Lim, K. C., Yusoff, F. M., Karim, M., & Natrah, F. M. I. (2023). Carotenoids modulate stress tolerance and immune responses in aquatic animals. Reviews in Aquaculture, 15(2), 872–894. https://doi.org/10.1111/raq.12767
  18. Oros, A., Coatu, V., Damir, N., Danilov, D., Ristea, E., & Lazar, L. (2025). Molecular mechanisms and biomarker-based early-warning indicators of heavy metal toxicity in marine fish. Fishes, 10(7), 339. https://doi.org/10.3390/fishes10070339
  19. Gonzбlez-Peсa, M. A., Ortega-Regules, A. E., Anaya de Parrodi, C., & Lozada-Ramнrez, J. D. (2023). Chemistry, occurrence, properties, applications, and encapsulation of carotenoids – a review. Plants, 12(2), 313. https://doi.org/10.3390/plants12020313 
  20. Terao, J. (2023). Revisiting carotenoids as dietary antioxidants for human health and disease prevention. Food & Function, 14(17), 7799–7824. https://doi.org/10.1039/D3FO02330C
  21. Miazek, K., Beton, K., Śliwińska, A., & Brożek-Płuska, B. (2022). The effect of β-carotene, tocopherols and ascorbic acid as anti-oxidant molecules on human and animal in vitro/in vivo studies: a review of research design and analytical techniques used. Biomolecules, 12(8), 1087. https://doi.org/10.3390/biom12081087
  22. Muzyka, L. V., Dmytrenko, I. M., Slidzevskyi, B. L., & Kyrychuk, H. Ye. (2014). Osoblyvosti khronichnoi dii nyzkykh kontsentratsii fenolu na vmist β-karotynu v hepatopankreasi Lymnaea stagnalis [Peculiarities of the chronic action of low phenol concentrations on the content of β-carotene in the hepatopancreas of Lymnaea stagnalis]. In Biolohichni doslidzhennia-2014:Zbirnyk naukovykh prats V Vseukrainskoi naukovo-praktychnoi konferentsii molodykh uchenykh i studentiv [Biological Research 2014. Proceedings of the 5th All-Ukrainian Scientific and Practical Conference of Young Scientists and Students], 246–248.
  23. Kyrychuk, G. E., & Muzyka, L. V. (2015). The effect of ammonium chloride concentrations on the content of carotenoid pigments in the body of Lymnaea stagnalis. Biosystems Diversity, 23(2), 154–160. https://doi.org/10.15421/011522 
  24. Kyrychuk, G. Ye., & Muzyka, L. V. (2016). Peculiarities of the distribution of β-carotene in the organism of Lymnaea stagnalis under the influence of the ions of heavy metals. Hydrobiological Journal, 52(5), 63–72. https://doi.org/10.1615/HydrobJ.v52.i5.70
  25. Kyrychuk, G. Ye., Muzyka, L. V., & Mykula, M. M. (2024). Peculiarities of low concentrations of heavy metal ions’ effect on β-carotene content in Lymnaea stagnalis. Hydrobiological Journal, 60(4), 72–84. https://doi.org/10.1615/HydrobJ.v60.i4.50
  26. Campoy-Diaz, A. D., Malanga, G., Giraud-Billoud, M., & Vega, I. A. (2023). Changes in the oxidative status and damage by non-essential elements in the digestive gland of the gastropod Pomacea canaliculata. Frontiers in Physiology, 14, 1123977. https://doi.org/10.3389/fphys.2023.1123977 
  27. Calabrese, E. J., & Mattson, M. P. (2017). How does hormesis impact biology, toxicology, and medicine? NPJ Aging and Mechanisms of Disease, 3(1), 13. https://doi.org/10.1038/s41514-017-0013-z
  28. Harrison, P. J., & Bugg, T. D. (2014). Enzymology of the carotenoid cleavage dioxygenases: reaction mechanisms, inhibition and biochemical roles. Archives of Biochemistry and Biophysics, 544, 105–111. https://doi.org/10.1016/j.abb.2013.10.005
  29. Somuncu, S., Atmaca, H., & Ilhan, S. (2024). Effects of acute exposure to environmentally realistic tebuconazole concentrations on stress responses of kidney and digestive gland of Lymnaea stagnalis. Environmental Toxicology and Pharmacology, 105, 104352. https://doi.org/10.1016/j.etap.2023.104352
  30. Boisseaux, P., Delignette-Muller, M. L., Abbaci, K., Thomas, H., & Garric, J. (2016). Analysis of hemocytes in Lymnaea stagnalis: characterization and effects of repeated hemolymph collections. Fish & Shellfish Immunology, 57, 116–126. https://doi.org/10.1016/j.fsi.2016.08.007 
  31. Livingstone, D. R. (2001). Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Marine Pollution Bulletin, 42(8), 656–666. https://doi.org/10.1016/S0025-326X(01)00060-1
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