Effects of spirulina (Arthrospira maxima) on teratogenicity and diclofenac-induced oxidative damage in Xenopus laevis

Abstract

Diclofenac (DCF) is a medication that is highly consumed and eliminated worldwide; it is constantly detected in the environment (primarily in water) and resists conventional degradation processes. It was included in the European Union watch list for the water framework. There are no regulations for this compound in Mexico. Therefore, this study evaluated the protective effect antioxidant activity of spirulina (Arthrospira maxima) against DCF-induced toxicity in Xenopus laevis at early life stages. X. laevis oocytes were exposed at the medium blastula stage for 96 h to three different mixtures: DCF+S 2 (149 µg L^-1 DCF plus 2 mg L^-1 spirulina), DCF+S 4 (149 µg L^-1 DCF plus 4 mg L^-1 spirulina), DCF+S 10 (149 µg L^-1 DCF plus 10 mg L^-1 spirulina). Other groups of oocytes were also exposed to DCF 149 µg L^-1 and a control group. The mortality and malformation rate, growth, lipid peroxidation, and antioxidant enzymatic activity (superoxide dismutase and catalase) were determined. Spirulina at 4 and 10 mg L^-1 reduced DCF-induced mortality by 80% and reduced malformations in severity and frequency. The abnormalities were malformations of the eye, tail, notochord, intestine, and rectum. All spirulina exposure groups showed an increase in total body size compared to those exposed to DCF. Regarding oxidative damage, the groups exposed to the mixture with spirulina decreased lipid peroxidation levels and diminished antioxidant activity. Spirulina reduced DCF-induced damage in X. laevis at early life stages and decreased mortality, frequency, and severity of abnormalities, growth inhibition, and oxidative damage. Further research is needed to evaluate the effects of spirulina against toxicity induced by xenobiotics in the early stages of development.

Keywords

Pharmaceuticals / diclofenac / spirulina / teratogenesis / oxidative stress / Xenopus laevis

References

[1] Stefanakis AI.A review of emerging contaminants in water. In: Mckeown AE, Bugyi G, editors. Impact of Water Pollution on Human Health and Environmental Sustainability. IGI Global; 2016. pp. 55-80.

[2] Ajima MN,Audu BS.Chronic diclofenac (DCF) exposure alters both enzymatic and haematological profile of African catfish, Clarias gariepinus.Drug Chem Toxicol2015;38:383-90

[3] Fekadu S,Dewil R.Pharmaceuticals in freshwater aquatic environments: a comparison of the African and European challenge.Sci Total Environ2019;654:324-37

[4] Hanif H,Kali S.Environmental risk assessment of diclofenac residues in surface waters and wastewater: a hidden global threat to aquatic ecosystem.Environ Monit Assess2020;192:204

[5] Melvin SD.Oxidative stress, energy storage, and swimming performance of Limnodynastes peronii tadpoles exposed to a sub-lethal pharmaceutical mixture throughout development.Chemosphere2016;150:790-7

[6] Gómez-Oliván LM,García-Medina S,Islas-Flores H.Genotoxic response and oxidative stress induced by diclofenac, ibuprofen and naproxen in Daphnia magna.Drug Chem Toxicol2014;37:391-9

[7] Gan TJ.Diclofenac: an update on its mechanism of action and safety profile.Curr Med Res Opin2010;26:1715-31

[8] Ulubay M,Kaplan AA.The use of diclofenac sodium in urological practice: a structural and neurochemical based review.J Chem Neuroanat2018;87:32-6

[9] Altman R,Brune K,Young C.Advances in NSAID development: evolution of diclofenac products using pharmaceutical technology.Drugs2015;75:859-77 PMCID:PMC4445819

[10] Bickley LK,Brown AR.Bioavailability and kidney responses to diclofenac in the fathead minnow (pimephales promelas).Environ Sci Technol2017;51:1764-74

[11] Cunha SC,Fernandes JO.Mussels as bioindicators of diclofenac contamination in coastal environments.Environ Pollut2017;225:354-60

[12] Bonnefille B,Courant F,Fenet H.Diclofenac in the marine environment: a review of its occurrence and effects.Mar Pollut Bull2018;131:496-506

[13] Sathishkumar P,Palanisami T,Palvannan T.Occurrence, interactive effects and ecological risk of diclofenac in environmental compartments and biota - a review.Sci Total Environ2020;698:134057

[14] Lonappan L,Das RK,Surampalli RY.Diclofenac and its transformation products: Environmental occurrence and toxicity - a review.Environ Int2016;96:127-38

[15] Bio S.Acute effects of diclofenac on zebrafish: Indications of oxidative effects and damages at environmentally realistic levels of exposure.Environ Toxicol Pharmacol2020;78:103394

[16] Yang YY,Liu YS.Pharmaceuticals and personal care products (PPCPs) and artificial sweeteners (ASs) in surface and ground waters and their application as indication of wastewater contamination.Sci Total Environ2018;616-617:816-23

[17] Kermia AEB,Trari M.Occurrence, fate and removal efficiencies of pharmaceuticals in wastewater treatment plants (WWTPs) discharging in the coastal environment of Algiers.Comptes Rendus Chimie2016;19:963-70

[18] Sousa DNR, Mozeto AA, Carneiro RL, Fadini PS. Spatio-temporal evaluation of emerging contaminants and their partitioning along a Brazilian watershed.Environ Sci Pollut Res Int2018;25:4607-20

[19] Branchet P,Fenet H.Anthropic impacts on Sub-Saharan urban water resources through their pharmaceutical contamination (Yaoundé, Center Region, Cameroon).Sci Total Environ2019;660:886-98

[20] Gumbi BP,Birungi G.Detection and quantification of acidic drug residues in South African surface water using gas chromatography-mass spectrometry.Chemosphere2017;168:1042-50

[21] González-Alonso S,Esteban S.Occurrence of pharmaceutical, recreational and psychotropic drug residues in surface water on the northern Antarctic Peninsula region.Environ Pollut2017;229:241-54

[22] Sharma BM,Scheringer M.Health and ecological risk assessment of emerging contaminants (pharmaceuticals, personal care products, and artificial sweeteners) in surface and groundwater (drinking water) in the Ganges River Basin, India.Sci Total Environ2019;646:1459-67

[23] Jindal K,Singh S.A systematic strategy for the identification and determination of pharmaceuticals in environment using advanced LC-MS tools: application to ground water samples.J Pharm Biomed Anal2015;108:86-96

[24] Kapelewska J,Karpińska J,Arciszewska A.Occurrence, removal, mass loading and environmental risk assessment of emerging organic contaminants in leachates, groundwaters and wastewaters.Microchem J2018;137:292-301

[25] Rodil R,Concha-Graña E,Muniategui-Lorenzo S.Emerging pollutants in sewage, surface and drinking water in Galicia (NW Spain).Chemosphere2012;86:1040-9

[26] Tröger R,Ahrens L.Micropollutants in drinking water from source to tap - method development and application of a multiresidue screening method.Sci Total Environ2018;627:1404-32

[27] Simazaki D,Suzuki T,Nishimura T.Occurrence of selected pharmaceuticals at drinking water purification plants in Japan and implications for human health.Water Res2015;76:187-200

[28] Carmona E,Picó Y.Occurrence of acidic pharmaceuticals and personal care products in Turia River Basin: from waste to drinking water.Sci Total Environ2014;484:53-63

[29] Brumovský M,Kohoutek J,Nizzetto L.Contaminants of emerging concern in the open sea waters of the Western Mediterranean.Environ Pollut2017;229:976-83

[30] Kallenborn R,Reiersen LO.Pharmaceuticals and personal care products (PPCPs) in Arctic environments: indicator contaminants for assessing local and remote anthropogenic sources in a pristine ecosystem in change.Environ Sci Pollut Res Int2018;25:33001-13

[31] Bayen S,Desai MM,Kelly BC.Occurrence and distribution of pharmaceutically active and endocrine disrupting compounds in Singapore’s marine environment: influence of hydrodynamics and physical-chemical properties.Environ Pollut2013;182:1-8

[32] Chernova E,Berezina N.Occurrence of pharmaceuticals in the Eastern Gulf of Finland (Russia).Environ Sci Pollut Res Int2021;28:68871-84

[33] Ali AM,Alarif W,Al-Lihaibi SS.Occurrence of pharmaceuticals and personal care products in effluent-dominated Saudi Arabian coastal waters of the Red Sea.Chemosphere2017;175:505-13

[34] Biel-Maeso M,Corada-Fernández C.Occurrence, distribution and environmental risk of pharmaceutically active compounds (PhACs) in coastal and ocean waters from the Gulf of Cadiz (SW Spain).Sci Total Environ2018;612:649-59

[35] Liu HQ,Li WW,Lam PKS.Spatial distribution and removal performance of pharmaceuticals in municipal wastewater treatment plants in China.Sci Total Environ2017;586:1162-9

[36] Lindholm-Lehto PC,Knuutinen JS.Widespread occurrence and seasonal variation of pharmaceuticals in surface waters and municipal wastewater treatment plants in central Finland.Environ Sci Pollut Res Int2016;23:7985-97

[37] Wilkinson JL,Hooda PS,Barton S.Markers of anthropogenic contamination: a validated method for quantification of pharmaceuticals, illicit drug metabolites, perfluorinated compounds, and plasticisers in sewage treatment effluent and rain runoff.Chemosphere2016;159:638-46

[38] Česen M,Terzić S,Heath E.The occurrence of contaminants of emerging concern in Slovenian and Croatian wastewaters and receiving Sava river.Sci Total Environ2019;650:2446-53

[39] Chiffre A,Buleté A,Badot PM.Occurrence of pharmaceuticals in WWTP effluents and their impact in a karstic rural catchment of Eastern France.Environ Sci Pollut Res Int2016;23:25427-41

[40] Christou A,Hapeshi E,Fatta-Kassinos D.Long-term wastewater irrigation of vegetables in real agricultural systems: concentration of pharmaceuticals in soil, uptake and bioaccumulation in tomato fruits and human health risk assessment.Water Res2017;109:24-34

[41] Corada-Fernández C,Candela L,Lara-Martín PA.Occurrence and spatial distribution of emerging contaminants in the unsaturated zone. case study: Guadalete River basin (Cadiz, Spain).Chemosphere2015;119 Suppl:S131-7

[42] Ashfaq M,Saif Ur Rehman M.Ecological risk assessment of pharmaceuticals in the receiving environment of pharmaceutical wastewater in Pakistan.Ecotoxicol Environ Saf2017;136:31-9

[43] Grossberger A,Borch T.Biodegradability of pharmaceutical compounds in agricultural soils irrigated with treated wastewater.Environ Pollut2014;185:168-77

[44] Čelić M,Farré M,Petrović M.Pharmaceuticals as chemical markers of wastewater contamination in the vulnerable area of the Ebro Delta (Spain).Sci Total Environ2019;652:952-63

[45] Peng FJ,Zhang M.Occurrence and ecological risk assessment of emerging organic chemicals in urban rivers: Guangzhou as a case study in China.Sci Total Environ2017;589:46-55

[46] Omar TFT,Yusoff FM.Occurrence, distribution, and sources of emerging organic contaminants in tropical coastal sediments of anthropogenically impacted Klang River estuary, Malaysia.Mar Pollut Bull2018;131:284-93

[47] Wilkinson JL,Swinden J,Barton S.Spatial distribution of organic contaminants in three rivers of Southern England bound to suspended particulate material and dissolved in water.Sci Total Environ2017;593-594:487-97

[48] Yan Q,Chen YP.Occurrence, fate and ecotoxicological assessment of pharmaceutically active compounds in wastewater and sludge from wastewater treatment plants in Chongqing, the Three Gorges Reservoir Area.Sci Total Environ2014;470-471:618-30

[49] Xue W,Xiao K.Elimination and fate of selected micro-organic pollutants in a full-scale anaerobic/anoxic/aerobic process combined with membrane bioreactor for municipal wastewater reclamation.Water Res2010;44:5999-6010

[50] Stasinakis AS,Arvaniti OS.Contribution of primary and secondary treatment on the removal of benzothiazoles, benzotriazoles, endocrine disruptors, pharmaceuticals and perfluorinated compounds in a sewage treatment plant.Sci Total Environ2013;463-464:1067-75

[51] Martín J,Aparicio I.Pharmaceutically active compounds in sludge stabilization treatments: anaerobic and aerobic digestion, wastewater stabilization ponds and composting.Sci Total Environ2015;503-504:97-104

[52] Rodríguez-Navas C,Bak SA.Pollution pathways of pharmaceutical residues in the aquatic environment on the island of Mallorca, Spain.Arch Environ Contam Toxicol2013;65:56-66

[53] Lu MC,Chiou MR,Fan HJ.Occurrence and treatment efficiency of pharmaceuticals in landfill leachates.Waste Manag2016;55:257-64

[54] Oliveira LL,Gonçalves F,Nunes B.Evaluation of ecotoxicological effects of drugs on Daphnia magna using different enzymatic biomarkers.Ecotoxicol Environ Saf2015;119:123-31

[55] Praskova E,Chromcova L.Effects of subchronic exposure of diclofenac on growth, histopathological changes, and oxidative stress in zebrafish (Danio rerio).Sci World J2014;2014:645737 PMCID:PMC3933404

[56] Peltzer PM,Martinuzzi C,Curi LM.Biotoxicity of diclofenac on two larval amphibians: assessment of development, growth, cardiac function and rhythm, behavior and antioxidant system.Sci Total Environ2019;683:624-37

[57] Memmert U,Burri R.Diclofenac: New data on chronic toxicity and bioconcentration in fish.Environ Toxicol Chem2013;32:442-52 PMCID:PMC3674524

[58] Nava-álvarez R,García-medina S,Galar-martínez M.Oxidative stress induced by mixture of diclofenac and acetaminophen on common carp (Cyprinus carpio).Water Air Soil Pollut2014;225

[59] Saucedo-Vence K,López-Martínez LX.Short and long-term exposure to diclofenac alter oxidative stress status in common carp Cyprinus carpio.Ecotoxicology2015;24:527-39

[60] Almeida Â,Soares AMVM.Anti-inflammatory drugs in the marine environment: bioconcentration, metabolism and sub-lethal effects in marine bivalves.Environ Pollut2020;263:114442

[61] Cardoso-Vera JD,SanJuan-Reyes N.Comparative study of diclofenac-induced embryotoxicity and teratogenesis in Xenopus laevis and Lithobates catesbeianus, using the frog embryo teratogenesis assay: Xenopus (FETAX).Sci Total Environ2017;574:467-75

[62] Islas-Flores H,Galar-Martínez M,Neri-Cruz N.Diclofenac-induced oxidative stress in brain, liver, gill and blood of common carp (Cyprinus carpio).Ecotoxicol Environ Saf2013;92:32-8

[63] Qureshi MA.Spirulina platensis exposure enhances macrophage phagocytic function in cats.Immunopharmacol Immunotoxicol1996;18:457-63

[64] Hayashi O,Okuwaki Y.Enhancement of antibody production in mice by dietary Spirulina platensis.J Nutr Sci Vitaminol1994;40:431-41

[65] Khan M,Mohan IK,Prayag A.Spirulina attenuates cyclosporine-induced nephrotoxicity in rats.J Appl Toxicol2006;26:444-51

[66] Karadeniz A,Simsek N.The effects of Panax ginseng and Spirulina platensis on hepatotoxicity induced by cadmium in rats.Ecotoxicol Environ Saf2009;72:231-5

[67] Kulshreshtha A,Jarouliya U,Prasad GB.Spirulina in health care management.Curr Pharm Biotechnol2008;9:400-5

[68] Parikh P,Iyer U.Role of spirulina in the control of glycemia and lipidemia in type 2 diabetes mellitus.J Med Food2001;4:193-9

[69] Sharma MK,Kumar A.Evaluation of protective efficacy of Spirulina fusiformis against mercury induced nephrotoxicity in Swiss albino mice.Food Chem Toxicol2007;45:879-87

[70] Rojas-Franco P,Blas-Valdivia V,Cano-Europa E.Arthrospira maxima (Spirulina) prevents endoplasmic reticulum stress in the kidney through its C-phycocyanin.J Zhejiang Univ Sci B2021;22:603-8 PMCID:PMC8284088

[71] Al-Qahtani WH.Anti-inflammatory, antioxidant and antihepatotoxic effects of Spirulina platensis against d-galactosamine induced hepatotoxicity in rats.Saudi J Biol Sci2019;26:647-52 PMCID:PMC6486513

[72] Lu J,Wang JZ,Egashira Y.Protection by dietary Spirulina platensis against D-galactosamine--and acetaminophen-induced liver injuries.Br J Nutr2010;103:1573-6

[73] James R,Nagarajan R,Manikandan MM.Effect of dietary Spirulina on reduction of copper toxicity and improvement of growth, blood parameters and phosphatases activities in carp, Cirrhinus mrigala (Hamilton, 1822).Indian J Exp Biol2009;47:754-9.

[74] Gad AS,El-Nekeety AA,Hassan NS.Antioxidant activity and hepatoprotective effects of whey protein and Spirulina in rats.Nutrition2011;27:582-9

[75] Zhang Y,Tang Q.The protective effects of selenium-enriched spirulina on the reproductive system of male zebrafish (Danio rerio) exposed to beta-cypermethrin.Food Funct2018;9:5791-804

[76] Tenorio-Chávez P,Gómez-Oliván LM.Chronic intake of an enriched diet with spirulina (Arthrospira maxima) alleviates the embryotoxic effects produced by realistic concentrations of tetracycline in Danio rerio.Sci Total Environ2023;859:159731

[77] Kim MY,Lee JH,Sok DE.Spirulina improves antioxidant status by reducing oxidative stress in rabbits fed a high-cholesterol diet.J Med Food2010;13:420-6

[78] Afkhami-Ardakani M,Shahrooz R,Malekinejad H.Antioxidant effects of Spirulina platensis (Arthrospira platensis) on cyclophosphamide-induced testicular injury in rats.Vet Res Forum2018;9:35-41 PMCID:PMC5913559

[79] Muga MA.Effects of fish oil and spirulina on oxidative stress and inflammation in hypercholesterolemic hamsters.BMC Complement Altern Med2014;14:470 PMCID:PMC4295402

[80] Toughan H,El-Ghoneimy AA,Seddek AS.Effect of dietary supplementation with Spirulina platensis on Atrazine-induced oxidative stress- mediated hepatic damage and inflammation in the common carp (Cyprinus carpio L.).Ecotoxicol Environ Saf2018;149:135-42

[81] Abdelkhalek NKM,Ahmed E.Protective role of dietary Spirulina platensis against diazinon-induced Oxidative damage in Nile tilapia; Oreochromis niloticus.Environ Toxicol Pharmacol2017;54:99-104

[82] Ael-D, Elbaghdady HA, Zahran E. Arsenic-induced genotoxicity in Nile tilapia (Orechromis niloticus); the role of Spirulina platensis extract.Environ Monit Assess2015;187:751

[83] Rajasekar P,Anjali R.Isolation and structural characterization of sulfated polysaccharide from Spirulina platensis and its bioactive potential: In vitro antioxidant, antibacterial activity and Zebrafish growth and reproductive performance.Int J Biol Macromol2019;141:809-21

[84] American Society for Testing Materials. Standard guide for conducting the frog embryo teratogenesis assay - Xenopus (FETAX). Available from: https://www.astm.org/e1439-12r19.html [Last accessed on 29 Mar 2023]

[85] Chae JP,Hwang YS.Evaluation of developmental toxicity and teratogenicity of diclofenac using Xenopus embryos.Chemosphere2015;120:52-8

[86] Pérez-Alvarez I,Gómez-Oliván LM,Chamorro-Cevallos G.Protective effects of Spirulina (Arthrospira maxima) against toxicity induced by cadmium in Xenopus laevis.Comp Biochem Physiol C Toxicol Pharmacol2021;248:109099

[87] Bantle JA,Finch RA. Atlas of abnormalities: a guide for the performance of FETAX. Stillwater, Okla: Printing Services, Oklahoma State University; 1991.

[88] Ishibashi S.How to grow Xenopus Laevis tadpole stages to adult.Cold Spring Harb Protoc2021;2021:pdb.prot106245

[89] Nieuwkoop PD.Normal table of Xenopus laevis (Daudin). A systematical and Chronological Survey of the Development from the Fertilized egg till the end of Metamorphosis, 1st ed. New York: Garland Publishing Inc; 1994.

[90] Bradford MM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal Biochem1976;72:248-54

[91] Buege JA.Microsomal lipid peroxidation.Methods Enzymol1978;52:302-10

[92] Misra HP.The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase.J Biol Chem1972;247:3170-5

[93] Radi R,Chang LY,Crapo JD.Detection of catalase in rat heart mitochondria.J Biol Chem1991;266:22028-34

[94] Miarov O,Avisar D.A critical evaluation of comparative regulatory strategies for monitoring pharmaceuticals in recycled wastewater.J Environ Manage2020;254:109794

[95] Tixier C,Oellers S.Occurrence and fate of carbamazepine, clofibric acid, diclofenac, ibuprofen, ketoprofen, and naproxen in surface waters.Environ Sci Technol2003;37:1061-8

[96] Petrie B,Kasprzyk-Hordern B.A review on emerging contaminants in wastewaters and the environment: current knowledge, understudied areas and recommendations for future monitoring.Water Res2015;72:3-27

[97] Bernadac Villegas LG,Carrillo-Méndez JD. Identificación y cuantificación de diclofenaco en aguas residuales de Ciudad Juárez. Available from: https://revista.itson.edu.mx/index.php/rlrn/article/view/281/214 [Last accessed on 29 Mar 2023]

[98] Stepanova S,Chromcova L.The effects of diclofenac on early life stages of common carp (Cyprinus carpio).Environ Toxicol Pharmacol2013;35:454-60

[99] Penha LCC,Martinez CBDR.Effects of anti-inflammatory diclofenac assessed by toxicity tests and biomarkers in adults and larvae of Danio rerio.Comp Biochem Physiol C Toxicol Pharmacol2021;242:108955

[100] Ajima MNO,Poojary N.Oxidative stress biomarkers, biochemical responses and Na⁺-K⁺-ATPase activities in Nile tilapia, Oreochromis niloticus exposed to diclofenac.Comp Biochem Physiol C Toxicol Pharmacol2021;240:108934

[101] Jung SH,Park SH.Diclofenac impairs autophagic flux via oxidative stress and lysosomal dysfunction: implications for hepatotoxicity.Redox Biol2020;37:101751 PMCID:PMC7575798

[102] Syed M,Hansen SH.Mitochondrial toxicity of diclofenac and its metabolites via inhibition of oxidative phosphorylation (ATP synthesis) in rat liver mitochondria: possible role in drug induced liver injury (DILI).Toxicol In Vitro2016;31:93-102

[103] Mirzaee SA,Ahmadi M.The possible oxidative stress and DNA damage induced in Diclofenac-exposed non-target organisms in the aquatic environment: a systematic review.Ecol Indic2021;131:108172

[104] Ghosh R,Feitoza LFBB,Gomes AV.Diclofenac induces proteasome and mitochondrial dysfunction in murine cardiomyocytes and hearts.Int J Cardiol2016;223:923-35 PMCID:PMC5076377

[105] Ramachandran A,Duan L,Jaeschke H.Mitochondrial dysfunction as a mechanism of drug-induced hepatotoxicity: current understanding and future perspectives.J Clin Transl Res2018;4:75-100 PMCID:PMC6261533

[106] Soni RA,Rana R.Spirulina – From growth to nutritional product: a review.2017;69:157-71

[107] Wu Q,Miron A,Wan D.The antioxidant, immunomodulatory, and anti-inflammatory activities of Spirulina: an overview.Arch Toxicol2016;90:1817-40

[108] Abu-Taweel GM,Antonisamy P.Spirulina consumption effectively reduces anti-inflammatory and pain related infectious diseases.J Infect Public Health2019;12:777-82

[109] Khafaga AF.Spirulina ameliorates methotrexate hepatotoxicity via antioxidant, immune stimulation, and proinflammatory cytokines and apoptotic proteins modulation.Life Sci2018;196:9-17

[110] Kumar P,Dwivedi M. Multiple potential roles of Spirulina in human health: a critical review. Available from: https://nutriweb.org.my/mjn/publication/21-3/j.pdf [Last accessed on 29 Mar 2023]

[111] Liang Y,Gao X.Effects of spirulina supplementation on lipid metabolism disorder, oxidative stress caused by high-energy dietary in Hu sheep.Meat Sci2020;164:108094

[112] Pestana JM,Santos H.Impact of dietary incorporation of Spirulina (Arthrospira platensis) and exogenous enzymes on broiler performance, carcass traits, and meat quality.Poult Sci2020;99:2519-32

[113] Slaby S,Marchand G.Maturation of Xenopus laevis oocytes under cadmium and lead exposures: cell biology investigations.Aquat Toxicol2017;193:105-10

[114] Aladaileh SH,Abd El-Hack ME.Spirulina platensis ameliorates the sub chronic toxicities of lead in rabbits via anti-oxidative, anti- inflammatory, and immune stimulatory properties.Sci Total Environ2020;701:134879

[115] Nasirian F,Moradi-Kor N.Effects of Spirulina platensis microalgae on antioxidant and anti-inflammatory factors in diabetic rats.Diabetes Metab Syndr Obes2018;11:375-80 PMCID:PMC6074810

[116] Banji D,Pratusha NG.Investigation on the role of Spirulina platensis in ameliorating behavioural changes, thyroid dysfunction and oxidative stress in offspring of pregnant rats exposed to fluoride.Food Chem2013;140:321-31

[117] Khalil SR,Awad A.Efficacy of Spirulina platensis diet supplements on disease resistance and immune-related gene expression in Cyprinus carpio L. exposed to herbicide atrazine.Fish Shellfish Immunol2017;67:119-28

[118] Hedayatirad M,Nematollahi MA,Brown C.Transgenerational disrupting impacts of atrazine in zebrafish: Beneficial effects of dietary spirulina.Comp Biochem Physiol C Toxicol Pharmacol2020;230:108685

[119] Abdelkhalek NKM,Ahmed E.Protective role of dietary Spirulina platensis against diazinon-induced Oxidative damage in Nile tilapia; Oreochromis niloticus.Environ Toxicol Pharmacol2017;54:99-104

[120] Bashandy SAE,Ebaid H.Antioxidant potential of Spirulina platensis mitigates oxidative stress and reprotoxicity induced by sodium arsenite in male rats.Oxid Med Cell Longev2016;2016

[121] El-Tantawy WH.Antioxidant effects of Spirulina supplement against lead acetate-induced hepatic injury in rats.J Tradit Complement Med2016;6:327-31 PMCID:PMC5067858

[122] Argüelles-Velázquez N,Madrigal-Bujaidar E.Amelioration of cadmium-produced teratogenicity and genotoxicity in mice given arthrospira maxima (Spirulina) treatment.Evid Based Complement Alternat Med2013;2013:604535 PMCID:PMC3863560

[123] Peter SJ,Giridharan R,Sabina EP.Suppressive effect of Spirulina fusiformis on diclofenac-induced hepato-renal injury and gastrointestinal ulcer in Wistar albino rats: a biochemical and histological approach.Biomed Pharmacother2017;88:11-8

[124] Park WS,Li M.Two classes of pigments, carotenoids and C-phycocyanin, in Spirulina powder and their antioxidant activities.Molecules2018;23:2065 PMCID:PMC6222893

[125] Bhat VB.C-phycocyanin: a potent peroxyl radical scavenger in vivo and in vitro.Biochem Biophys Res Commun2000;275:20-5

[126] Ch, González R, Ledón N, Remirez D, Rimbau V. C-phycocyanin: a biliprotein with antioxidant, anti-inflammatory and neuroprotective effects.Curr Protein Pept Sci2003;4:207-16

[127] Deng R.Hypolipidemic, antioxidant, and antiinflammatory activities of microalgae Spirulina.Cardiovasc Ther2010;28:e33-45 PMCID:PMC2907180

[128] Schafer FQ,Kelley EE,Martin SM.Comparing beta-carotene, vitamin E and nitric oxide as membrane antioxidants.Biol Chem2002;383:671-81

[129] Miyazawa T,Itaya M,Miyazawa T.Vitamin E: regulatory redox interactions.IUBMB Life2019;71:430-41

[130] Moradi A,Ghafourikhosroshahi A.Effects of vitamin A and vitamin E on attenuation of titanium dioxide nanoparticles-induced toxicity in the liver of male Wistar rats.Mol Biol Rep2019;46:2919-32

[131] Choi SW,Collins AR,Strain JJ.Vitamins C and E: acute interactive effects on biomarkers of antioxidant defence and oxidative stress.Mutat Res2004;551:109-17

[132] Ryan MJ,Docherty M.Vitamin E and C supplementation reduces oxidative stress, improves antioxidant enzymes and positive muscle work in chronically loaded muscles of aged rats.Exp Gerontol2010;45:882-95 PMCID:PMC3104015