The use of selected biomarkers, fatty acid and histopathology features to detect the possible toxic effects of the penconazole-containing fungicide Topas on marine clam siphons (Ruditapes decussatus)

Boutheina Ben Abdallah; Safa Bejaoui; Wafa Trabelsi; Dalya Belhassen; Zeineb Khila; Samir  Boubaker; Chayma  Ben Fayala; Nejla  Soudani

doi:10.3989/scimar.05562.095

Authors

Boutheina Ben Abdallah Laboratory of Ecology, Biology and Physiology of Aquatic Organisms (LR18ES41), Biology Department, Faculty of Sciences of Tunis, University of Tunis El Manar https://orcid.org/0000-0003-0139-4876
Safa Bejaoui Laboratory of Ecology, Biology and Physiology of Aquatic Organisms (LR18ES41), Biology Department, Faculty of Sciences of Tunis, University of Tunis El Manar https://orcid.org/0000-0002-7946-2763
Wafa Trabelsi Laboratory of Ecology, Biology and Physiology of Aquatic Organisms (LR18ES41), Biology Department, Faculty of Sciences of Tunis, University of Tunis El Manar https://orcid.org/0000-0002-1114-4080
Dalya Belhassen Laboratory of Ecology, Biology and Physiology of Aquatic Organisms (LR18ES41), Biology Department, Faculty of Sciences of Tunis, University of Tunis El Manar https://orcid.org/0009-0003-5518-1435
Zeineb Khila Laboratory of Ecology, Biology and Physiology of Aquatic Organisms (LR18ES41), Biology Department, Faculty of Sciences of Tunis, University of Tunis El Manar - Interdisciplinary Laboratory of Continental Environments, UMR 7360 LIEC CNRS University of Lorraine https://orcid.org/0000-0001-7603-1557
Samir Boubaker Pasteur Institute of Tunis, Department of Human and Experimental Anatomy Pathology https://orcid.org/0000-0001-5653-2936
Chayma Ben Fayala Pasteur Institute of Tunis, Department of Human and Experimental Anatomy Pathology https://orcid.org/0009-0003-1728-1988
Nejla Soudani Laboratory of Ecology, Biology and Physiology of Aquatic Organisms (LR18ES41), Biology Department, Faculty of Sciences of Tunis, University of Tunis El Manar - Department of Basic Sciences, Physiology and Functional Explorations Section, Faculty of Medicine of Tunis https://orcid.org/0000-0002-7652-9678

DOI:

https://doi.org/10.3989/scimar.05562.095

Keywords:

Ruditapes decussatus, siphons, antioxidant status, fatty acid profiling, histopathological features, Topas

Abstract

Widespread use of pesticides in agriculture has the potential to harm non-target creatures diffusely and contaminate aquatic habitats through leaching and runoff events. Triazoles are among the fungicides used extensively worldwide due to their efficacy against fungal crop diseases and their broad spectrum of action. In this study, the impact of Topas on the antioxidant defence system, fatty acid composition and histopathological injuries was experimentally examined under three concentrations (4, 40, and 400 µg L^-1) over 96 hours in Ruditapes decussatus siphons. Our results showed that Topas exposure induced a significant decrease in the levels of saturated fatty acids. However, an increase of monounsaturated fatty acids and polyunsaturated fatty acids, mainly the eicosapentaenoic (C20:5n-3), docosahexaenoic (C22:6n-3) and arachidonic (C20:4n-6) acids. Topas exposure enhanced the levels of hydrogen peroxide, malondialdehyde and protein carbonyls and altered enzymatic and non-enzymatic antioxidant status in all treated clams. Acetylcholinesterase activity was inhibited with the increase of Topas concentrations. Eventually, histopathological changes detected in treated animals varied in a concentration-dependent manner and were herein consistent with the biochemical outcomes. Our findings shed new light on the relationship between redox state and fatty acid composition changes, allowing us to understand Topas-triggered toxicity.

Downloads

Download data is not yet available.

References

Aebi H. 1984. Catalase in Vitro. Methods Enzymol. 105: 121-126. https://doi.org/10.1016/S0076-6879(84)05016-3 PMid:6727660

Ahammad Sahib I.K., Sailatha D., Ramana Rao K.V. 1980. Impact of malathion on acetylcholinesterase in the tissues of the fish Tilapia mossambica (Peters)-a time course study. J. Biosci. 2(1): 37-41. https://doi.org/10.1007/BF02703131

Alderman C.J.J., Shah S., Foreman J.C., et al. 2002. The role of advanced oxidation protein products in regulation of dendritic cell function. Free Radical. Biol. Med. 32(5): 377-385. https://doi.org/10.1016/S0891-5849(01)00735-3 PMid:11864777

Alkan Uçkun A., Barım Öz Ö. 2020. Acute exposure to the fungicide penconazole affects some biochemical parameters in the crayfish (Astacus leptodactylus Eschscholtz, 1823). Environ. Sci. Pollut. Res. 27(28): 35626-35637. https://doi.org/10.1007/s11356-020-09595-2 PMid:32601870

Ayala A., Muñoz M.F., Argüelles S. 2014. Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4- hydroxy-2-nonenal. Oxidative Med. Cell. Longev. 2014: 360438. https://doi.org/10.1155/2014/360438 PMid:24999379 PMCid:PMC4066722

Barhoumi B., Clérandeau C., Le Menach K., et al. 2014. Pollution biomonitoring in the Bizerte lagoon (Tunisia), using grass goby, Zosterisessor ophiocephalus (Teleostei, Gobiidae) as a bioindicator species. Mar. Environ. Res. 101: 184-195. https://doi.org/10.1016/j.marenvres.2014.07.002 PMid:25106667

Beauchamp C., Fridovich I. 1971. Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44(1): 276-287. https://doi.org/10.1016/0003-2697(71)90370-8 PMid:4943714

Benzie I.F.F., Strain J.J. 1996. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": The FRAP assay. Anal. Biochem. 239(1): 70-76. https://doi.org/10.1006/abio.1996.0292 PMid:8660627

Calder P.C. 2010. Omega-3 fatty acids and inflammatory processes. Nutrients 2(3): 355-374. https://doi.org/10.3390/nu2030355 PMid:22254027 PMCid:PMC3257651

Cecchi G., Biasini S.C.J. 1985. Methanolyse rapide des huiles en solvants. Note de Laboratoire. Rev. Franc. Corps. Gras. 4: 163-164.

Chaâbane M., Ghorbel I., Elwej A., et al. 2016. Penconazole alters redox status, cholinergic function, and membrane-bound ATPases in the cerebrum and cerebellum of adult rats. Hum. Exp. Toxicol. 36(8): 854-866. https://doi.org/10.1177/0960327116672911 PMid:27738198

Chaâbane M., Elwej A., Ghorbel I., Chelly S., Mnif H., Boudawara T., Ellouze chaabouni S., Zeghal N., Soudani, N. 2018. Penconazole alters redox status, cholinergic function and lung's histoarchitecture of adult rats: Reversal effect of vitamin E. Biomed. Pharmacother. 102: 645-652. https://doi.org/10.1016/j.biopha.2018.03.113 PMid:29602133

Chalghmi H., Bourdineaud J.P., Haouas Z., et al. 2016. Transcriptomic, Biochemical, and Histopathological Responses of the Clam Ruditapes decussatus from a Metal-Contaminated Tunis Lagoon. Arch. Environ. Contam. Toxicol. 70(2), 241-256. https://doi.org/10.1007/s00244-015-0185-0 PMid:26077924

Chetoui I., Bejaoui S., Trabelsi W., et al. 2019. Exposure of Mactra corallina to acute doses of lead: effects on redox status, fatty acid composition and histomorphological aspect. Drug Chem. Toxicol. 45(1): 311-323. https://doi.org/10.1080/01480545.2019.1693590 PMid:31752645

Cheung C.C.C., Zheng G.J., Li A.M.Y., et al. 2001. Relationships between tissue concentrations of polycyclic aromatic hydrocarbons and antioxidative responses of marine mussels, Perna viridis. Aquat. Toxicol. 52: 189-203. 4 https://doi.org/10.1016/S0166-445X(00)00145-4 PMid:11239681

Costa P.M., Carreira S., Costa M.H., et al. 2013. Development of histopathological indices in a commercial marine bivalve (Ruditapes decussatus) to determine environmental quality. Aquat. Toxicol. 126(2013): 442-454. https://doi.org/10.1016/j.aquatox.2012.08.013 PMid:23010389

Cravo A., Pereira C., Gomes T., et al. 2012. A multibiomarker approach in the clam Ruditapes decussatus to assess the impact of pollution in the Ria Formosa lagoon, South Coast of Portugal. Mar. Environ. Res. 75: 23-34. https://doi.org/10.1016/j.marenvres.2011.09.012 PMid:22001190

Dalvie M.A., Cairncross E., Solomon A., et al. 2003. Contamination of rural surface and ground water by endosulfan in farming areas of the Western Cape, South Africa. Environ Health. 2(1): 1. https://doi.org/10.1186/1476-069X-2-1 PMid:12689341 PMCid:PMC153526

De Vico G., Carella F. 2012. Morphological features of the inflammatory response in molluscs. Res. Vet. Sci. 93(3): 1109-1115. https://doi.org/10.1016/j.rvsc.2012.03.014 PMid:22513124

Ding F., Song W.H., Guo J., et al. 2009. Oxidative stress and structure-activity relationship in the zebrafish (Danio rerio) under exposure to paclobutrazol. J. Environ. Sci. Health. - Part B Pesticides, Food Contaminants, and Agricultural Wastes. 44(1): 44-50. https://doi.org/10.1080/03601230802519652 PMid:19089714

Draper H.H., Hadley M. 1990. Malondialdehyde determination as index of lipid Peroxidation. Methods. Enzymol. 186(C): 421-431. https://doi.org/10.1016/0076-6879(90)86135-I PMid:2233309

Ellman G.L. 1959. Tissue Sulfhydryl Groups. Arch. Biochem. Biophys. 82: 70-77. https://doi.org/10.1016/0003-9861(59)90090-6 PMid:13650640

Ellman G.L., Courtney K.D., Andres V. 1961. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7: 88-95. https://doi.org/10.1016/0006-2952(61)90145-9 PMid:13726518

European chemicals agency (ECHA) 2023. Retrieved 23 May 2024 from https://echa.europa.eu/substance-information/-/substanceinfo/100.060.231

Floch J., Lees M., Sloane Stanley G.H. 1957. A simple method for the isolation and purification of total lipides from animal tissues. J. Biol. Chem. 226(1): 497-509. https://doi.org/10.1016/S0021-9258(18)64849-5 PMid:13428781

Flohé L., Günzler W.A. 1984. Assays of Gluthathione Peroxidase. Methods. Enzymol. 105: 114-120. https://doi.org/10.1016/S0076-6879(84)05015-1 PMid:6727659

Fokina N.N., Ruokolainen T.R., Nemova N.N., et al. 2013. Changes of blue mussels Mytilus edulis L. lipid composition under cadmium and copper toxic effect. Biol. Trace. Elem. Res. 154(2): 217-225. https://doi.org/10.1007/s12011-013-9727-3 PMid:23793920

Fouzai C., Trabelsi W., Bejaoui S., et al. 2020a. Cellular toxicity mechanisms of lambda-cyhalothrin in Venus verrucosa as revealed by fatty acid composition, redox status and histopathological changes. Ecol. Indic. 108. https://doi.org/10.1016/j.ecolind.2019.105690

Fouzai C., Trabelsi W., Rabeh I., et al. 2020b. Fatty acid profile and antioxidant status in Venus verrucosa gills as λ-cyhalothrin toxic effects. Toxicol. Ind. Health. 36(11): 898-907. https://doi.org/10.1177/0748233720961219 PMid:32996380

Ghribi F., Richir J., Bejaoui S., et al. 2020. Trace elements and oxidative stress in the Ark shell Arca noae from a Mediterranean coastal lagoon (Bizerte lagoon, Tunisia): are there health risks associated with their consumption? Environ. Sci. Pollut. Res. 27(13): 15607-15623. https://doi.org/10.1007/s11356-020-07967-2 PMid:32128728

Grünberger O., Hamdi R., Lagacherie M., et al. 2024. Pesticide contamination pattern of surface water in an urban-agricultural mediterranean watershed (Wadi Guenniche, Bizerte Lagoon, Northern Tunisia). J. Environ. Sci. Health. Part B. 59(8): 521-539. https://doi.org/10.1080/03601234.2024.2375905 PMid:39001801

Habig W.H., Pabst M.J., Fleischner G., et al. 1974. The identity of glutathione S-transferase B with ligandin, a major binding protein of liver. Proc. Natl. Acad. Sci. 71(10): 3879-3882. https://doi.org/10.1073/pnas.71.10.3879 PMid:4139704 PMCid:PMC434288

Hamza-Chaffai A., Pellerin J., Amiard J.C. 2003. Health assessment of a marine bivalve Ruditapes decussatus from the Gulf of Gabès (Tunisia). Environ. Int. 28(7): 609-617. https://doi.org/10.1016/S0160-4120(02)00102-2 PMid:12504157

Hatami M., Banaee M., Nematdoost Haghi B. 2019. Sub-lethal toxicity of chlorpyrifos alone and in combination with polyethylene glycol to common carp (Cyprinus carpio). Chemosphere 219: 981-988. https://doi.org/10.1016/j.chemosphere.2018.12.077 PMid:30682763

Hart K.A., Pimentel D. 2002. Environmental and economic costs of pesticide use. In: Pimentel D, editor. Encyclopedia of pest management. New York: Marcel Dekker; p. 237-239. https://doi.org/10.1201/NOE0824706326.ch99

Hermsen S.A.B., Pronk T.E., van den Brandhof E.J., et al. 2011. Chemical class-specific gene expression changes in the zebrafish embryo after exposure to glycol ether alkoxy acids and 1,2,4-triazole antifungals. Reprod. Toxicol. 32(2): 245-252. https://doi.org/10.1016/j.reprotox.2011.05.010 PMid:21621602

Husak V.V., Mosiichuk N.M., Storey J.M., et al. 2017. Acute exposure to the penconazole-containing fungicide Topas partially augments antioxidant potential in goldfish tissues. Comp. Biochem. Physiol. Part - C. 193: 1-8. https://doi.org/10.1016/j.cbpc.2016.12.003 PMid:27979696

Icoglu Aksakal F., Ciltas A. 2018. Developmental toxicity of penconazole in Zebrfish (Danio rerio) embryos. Chemosphere. 200: 8-15. https://doi.org/10.1016/j.chemosphere.2018.02.094 PMid:29471168

Jacques-Silva M.C., Nogueira C.W., Broch L.C. 2001. Diphenyl diselenide and ascorbic acid changes deposition of selenium and ascorbic acid in liver and brain of mice. Pharmacol. Toxicol. 88: 119-125. https://doi.org/10.1034/j.1600-0773.2001.d01-92.x PMid:11245406

Jifa W., Yu Z., Xiuxian S., et al.2006. Response of integrated biomarkers of fish (Lateolabrax japonicus) exposed to benzo[a]pyrene and sodium dodecylbenzene sulfonate. Ecotoxicol. Environ. Saf. 65(2): 230-236. https://doi.org/10.1016/j.ecoenv.2005.08.002 PMid:16256195

Jollow D.J., Mitchell J., Zampaglione N., et al. 1974. Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3,4-bromobenzene oxide as the hepatotoxic metabolite. Pharmacology. (11): 151-169. https://doi.org/10.1159/000136485 PMid:4831804

Kalliora C., Mamoulakis C., Vasilopoulos E., et al. 2018. Association of pesticide exposure with human congenital abnormalities. Toxicol. Appl. Pharmacol. 346(March): 58-75. https://doi.org/10.1016/j.taap.2018.03.025 PMid:29596925 PMCid:PMC6029725

Kamel N., Burgeot T., Banni M., et al. 2014. Effects of increasing temperatures on biomarker responses and accumulation of hazardous substances in rope mussels (Mytilus galloprovincialis) from Bizerte lagoon. Environ. Sci. Pollut. Res. 21: 6108-6123 https://doi.org/10.1007/s11356-014-2540-5 PMid:24474562

Kayali R., Çakatay U., Akçay T., et al. 2006. Effect of alpha‐lipoic acid supplementation on markers of protein oxidation in post‐mitotic tissues of ageing rat. Cell Biochem. Funct: Cellular biochemistry and its modulation by active agents or disease. 24(1): 79-85. https://doi.org/10.1002/cbf.1190 PMid:15532093

Konwick B.J., Garrison A.W., Avants J.K., et al. 2006. Bioaccumulation and biotransformation of chiral triazole fungicides in rainbow trout (Oncorhynchus mykiss). Aquat. Toxicol. 80: 372-381. https://doi.org/10.1016/j.aquatox.2006.10.003 PMid:17118468

Krishnan N., Kodrík D., Kłudkiewicz B., et al. 2009. Glutathione-ascorbic acid redox cycle and thioredoxin reductase activity in the digestive tract of Leptinotarsa decemlineata (say). Insect. Biochem. Mol. Biol. 39: 180-188. https://doi.org/10.1016/j.ibmb.2008.11.001 PMid:19049872

Krumova K., Cosa G. 2016. Chapter 1: Overview of reactive oxygen species, in Singlet oxygen: applications in biosciences and nanosciences, Volume 1, pp. 1-21. https://doi.org/10.1039/9781782622208-00001

Li Z.H., Zlabek V., Grabic R., et al. 2010a. Effects of exposure to sublethal propiconazole on the antioxidant defense system and Na+-K+-ATPase activity in brain of rainbow trout, Oncorhynchus mykiss. Aquat. Toxicol. 98(3): 297-303. https://doi.org/10.1016/j.aquatox.2010.02.017 PMid:20363517

Li Z.H., Zlabek V., Li P., et al. 2010b. Biochemical and physiological responses in liver and muscle of rainbow trout after long-term exposure to propiconazole. Ecotoxicol. Environ. Saf. 73: 1391-1396. https://doi.org/10.1016/j.ecoenv.2010.05.017 PMid:20621356

Liu J.J., Green P., John-Mann J., et al. 2015. Pathways of polyunsaturated fatty acid utilization: Implications for brain function in neuropsychiatric health and disease. Brain. Res. 1597: 220-246. https://doi.org/10.1016/j.brainres.2014.11.059 PMid:25498862 PMCid:PMC4339314

Llesuy S., Evelson P., Campos A.M., et al. 2001. Methodologies for evaluation of total antioxidant activities in complex mixtures. A critical review. Biol. Res. 34(2): 51-73. https://doi.org/10.4067/S0716-97602001000200009 PMid:11715209

López Martínez R. 2024. Study of the establishment of an allocated zone for aquaculture (AZA) in the Lagoon of Bizerte, Tunisia. 112 pp.

Lowry O.H., Rosebrough N.J., Farr A.L., et al. 1951. Protein measurement with the Folin phenol reagent. J. Biolo. Chem. 193(1): 265-275. https://doi.org/10.1016/S0021-9258(19)52451-6 PMid:14907713

Mao H., Wang D.H., Yang W.X. 2012. The involvement of metallothionein in the development of aquatic invertebrate. Aquat. Toxicol. 110-111: 208-213. https://doi.org/10.1016/j.aquatox.2012.01.018 PMid:22343466

Martoja R., Martoja-Pierson M. 1967. Initiation aux techniques de l'histologie animale, Masson: Paris, 345.

Monroig Ó., Kabeya N. 2018. Desaturases and elongases involved in polyunsaturated fatty acid biosynthesis in aquatic invertebrates: a comprehensive review. Fisheries. Sci. 84(6): 911-928. https://doi.org/10.1007/s12562-018-1254-x

Monserrat J.M., Martinez P.E., Geracitano L.A., et al. 2007. Pollution biomarkers in estuarine animals: critical review and new perspectives. Comp. Biochem. Physiol. - Part C 146: 221-234. https://doi.org/10.1016/j.cbpc.2006.08.012 PMid:17045848

Mu X., Pang S., Sun X., et al. 2013. Evaluation of acute and developmental effects of difenoconazole via multiple stage zebrafish assays. Environ. Pollut. 175: 147-157. https://doi.org/10.1016/j.envpol.2012.12.029 PMid:23391686

Necibi M., Mzoughi N. 2020. Distribution of organochlorine pesticides in sediment cores from the Bizerte Lagoon (Tunisia). Int. J. of Environ. Anal. Chem. 100(10): 1118-1132. https://doi.org/10.1080/03067319.2019.1650173

Ou P., Wolff S.P. 1996. A discontinuous method for catalase determination at "near physiological" concentrations of H2O2 and its application to the study of H2O2 fluxes within cells. J. Biochem. Biophys. Methods. 31(1-2): 59-67. https://doi.org/10.1016/0165-022X(95)00039-T PMid:8926339

Pagano M., Capillo G., Sanfilippo M., et al. 2016. Evaluation of Functionality and Biological Responses of Mytilus galloprovincialis after Exposure to Quaternium-15 (Methenamine 3-Chloroallylochloride). Molecules. 21(2): 1-12. https://doi.org/10.3390/molecules21020144 PMid:26821003 PMCid:PMC6273939

Pamplona R. 2008. Membrane phospholipids, lipoxidative damage and molecular integrity: a causal role in aging and longevity. Biochim. Biophys. Acta. 1777(10): 1249-1262. https://doi.org/10.1016/j.bbabio.2008.07.003 PMid:18721793

Peffer R.C., Moggs J.G., Pastoor T., et al. 2007. Mouse liver effects of cyproconazole, a triazole fungicide: role of the constitutive androstane receptor. Toxicol. Sci. 99: 315-325. https://doi.org/10.1093/toxsci/kfm154 PMid:17557908

Petrović S., Ozretić B., Krajnović-Ozretić M., et al. 2001. Lysosomal membrane stability and metallothioneins in digestive gland of mussels (Mytilus galloprovincialis Lam.) as biomarkers in a field study. Mar. Pollut. Bull. 42(12): 1373-1378. https://doi.org/10.1016/S0025-326X(01)00167-9 PMid:11827125

Reznick A.Z., Packer L. 1994. Oxidative damage to proteins: Spectrophotometric method for carbonyl assay. Methods. Enzymol. 233: 357-363. https://doi.org/10.1016/S0076-6879(94)33041-7 PMid:8015470

Rodríguez-Fuentes G., Rubio-Escalante F.J., Noreña-Barroso E., et al. 2015. Impacts of oxidative stress on acetylcholinesterase transcription, and activity in embryos of zebrafish (Danio rerio) following Chlorpyrifos exposure. Comp. Biochem. Physiol Part - C: Toxicology and Pharmacology. 172-173: 19-25. https://doi.org/10.1016/j.cbpc.2015.04.003 PMid:25937383

Sanchez W., Piccini B., Porcher J.M. 2008. Effect of prochloraz fungicide on biotransformation enzymes and oxidative stress parameters in three-spined stickleback (Gasterosteus aculeatus L.). J. Environ. Sci. Health. B 43 (1): 65-70. https://doi.org/10.1080/03601230701735151 PMid:18161575

Sayeed I., Parvez S., Pandey S., et al. 2003. Oxidative stress biomarkers of exposure to deltamethrin in freshwater fish, Channa punctatus Bloch. Ecotoxicol. Environ. Saf. 56(2): 295-301. https://doi.org/10.1016/S0147-6513(03)00009-5 PMid:12927561

Signa G., Di Leonardo R., Vaccaro A., et al. 2015. Lipid and fatty acid biomarkers as proxies for environmental contamination in caged mussels Mytilus galloprovincialis. Ecol. Indic. 57: 384-394. https://doi.org/10.1016/j.ecolind.2015.05.002

Smii H., Khazri A., Ali M., et al. 2021. Titanium dioxide nanoparticles are toxic for the freshwater mussel Unio ravoisieri: Evidence from a multimarker approach. Diversity. 13(12): 1-17. https://doi.org/10.3390/d13120679

Stara A., Pagano M., Albano M., et al. 2021. Effects of long-term exposure of Mytilus galloprovincialis to thiacloprid: A multibiomarker approach. Environ. Pollut. 289(August): 117892. https://doi.org/10.1016/j.envpol.2021.117892 PMid:34385134

Surfrider. 2020. Chemical pollution of the ocean: the pesticide issue. Retrieved October 3, 2024, from https://www.surfrider.eu/learn/news/pollution-chimique-ocean-question-pesticides

Tallima H., El Ridi R. 2018. Arachidonic acid: Physiological roles and potential health benefits - A review. J. Adv. Res. 11: 33-41. https://doi.org/10.1016/j.jare.2017.11.004 PMid:30034874 PMCid:PMC6052655

Telahigue K., Rabeh I., Hajji T., et al. 2020. Assessment of the impacts of glyphosate and its commercial formulation Roundup® on the respiratory tree of the sea cucumber Holothuria forskali using a multivariate biomarker approach. Chemosphere. https://doi.org/10.1016/j.chemosphere.2020.129376 PMid:33385670

Trabelsi W., Chetoui I., Fouzai C., et al. 2019. Redox status and fatty acid composition of Mactra corallina digestive gland following exposure to acrylamide. Environ. Sci. Pollut. Res. 26: 22197-22208. https://doi.org/10.1007/s11356-019-05492-5 PMid:31148000

Ullah R., Zuberi A., Ullah S., et al. 2014. Cypermethrin induced behavioral and biochemical changes in mahseer, Tor putitora. J. Toxicol. Sci. 39: 829-836. https://doi.org/10.2131/jts.39.829 PMid:25374374

Uluturhan E., Darılmaz E., Kontas A., et al. 2019. Seasonal variations of multi-biomarker responses to metals and pesticides pollution in M. galloprovincialis and T. decussatus from Homa Lagoon, Eastern Aegean Sea. Mar. Pollut. Bull. 141: 176-186. https://doi.org/10.1016/j.marpolbul.2019.02.035 PMid:30955723

Viarengo A., Ponzano E., Dondero F., et al. 1997. A simple spectrophotometric method for metallothionein evaluation in marine organisms: An application to Mediterranean and Antarctic molluscs. Mar. Environ. Res. 44(1): 69-84. https://doi.org/10.1016/S0141-1136(96)00103-1

Wang C., Wu Q., Wu C., et al. 2011. Application of dispersion-solidification liquid-liquid microextraction for the determination of triazole fungicides in environmental water samples by high-performance liquid chromatography. J. Hazard. Mater. 185(1): 71-76. https://doi.org/10.1016/j.jhazmat.2010.08.124 PMid:20875927

Yin X., Chen P., Chen H., Jin W. Yan X. 2017. Physiological performance of the intertidal Manila clam (Ruditapes philippinarum) to long-term daily rhythms of air exposure. Sci. rep. 7: 1-12. https://doi.org/10.1038/srep41648 PMid:28128354 PMCid:PMC5269718

Yoloğlu E. 2019. Assessment of Na+/K+ -ATPase, Mg2+-ATPase, Ca2+-ATPase, and Total-ATPase Activities in Gills of Freshwater Mussels Exposed to Penconazole. Comm. J. Biol. 3: 88-92. https://doi.org/10.31594/commagene.632082

Zhukova N.V. 1991. The pathway of the biosynthesis of non-methylene-interrupted dienoic fatty acids in molluscs. Comp. Biochem. Physiol. 100(4): 801-804. https://doi.org/10.1016/0305-0491(91)90293-M