Abstract
The incorporation of macroalgae on aquafeeds has been suggested to increase fish growth and organoleptic characteristics. Moreover, the potential of macroalgae to strengthen fish capacity to cope with different stressors is a challenging topic in the field of applied phycology. The adverse side effects of aqua-medicines, combined with the hypothesis that a macroalgae-enriched diet can minimize that impact, are the starting point for the present study. Hence, a mix of Ulva rigida, Fucus vesiculosus and Gracilaria gracilis was incorporated (5%) in the gilthead seabream (Sparus aurata) diet, to evaluate a possible protection against the chromosomal damage (measured as erythrocytic nuclear abnormalities) induced by the antibiotic oxytetracycline and the antiparasitic formalin as well as by the model genotoxicant cyclophosphamide. A 60-day differential dietary background was implemented (one fish group fed with standard diet and other with macroalgae-supplemented diet), after which fish were challenged by oxytetracycline, formalin and cyclophosphamide, and appraisals carried out 4 and 18 days later. To shed light on the modulation of the erythrocytic population dynamics, the erythrocyte maturity index was assessed. Oxytetracycline and formalin displayed a chromosome-damaging potential that remained for 18 days. The macroalgae-enriched diet evidenced a solid genoprotection against the three agents, revealing a broad-spectrum action. Oxytetracycline and formalin induced an erythrocyte population instability (translated into an aging effect), counteracted by algae supplementation, which seems to convey erythropoiesis promoting factors. Overall, the benefits given by the algae supplementation recommend its adoption within a framework of sustainable aquaculture practices, namely as a prophylactic measure to mitigate productivity losses when oxytetracycline and formalin are used.
Similar content being viewed by others
References
AADAPP (2010) A quick reference guide to approved drugs for use in aquaculture. https://www.fws.gov/fisheries/aadap/PDF/approveddrugs.pdf (Accessed at 17/09/2019)
Ambili TR, Saravanan M, Ramesh M, Abhijith DB, Poopal RK (2013) Toxicological effects of the antibiotic oxytetracycline to an Indian major carp Labeo rohita. Arch Environ Contam Toxicol 64:494–503
Araújo M, Rema P, Sousa-Pinto I, Cunha LM, Peixoto MJ, Pires MA, Seixas F, Brotas V, Beltrán C, Valente LMP (2016) Dietary inclusion of IMTA-cultivated Gracilaria vermiculophylla in rainbow trout (Oncorhynchus mykiss) diets: effects on growth, intestinal morphology, tissue pigmentation, and immunological response. J Appl Phycol 28:679–689
Athukorala Y, Kim K-N, Jeon Y-J (2006) Antiproliferative and antioxidant properties of an enzymatic hydrolysate from brown alga, Ecklonia cava. Food Chem Toxicol 44:1065–1074
Ayllón F, Garcia-Vazquez E (2001) Micronuclei and other nuclear lesions as genotoxicity indicators in rainbow trout Oncorhynchus mykiss. Ecotoxicol Environ Saf 49:221–225
Barker S, Weinfeld M, Murray D (2005) DNA–protein crosslinks: their induction, repair, and biological consequences. Mutat Res Mutat Res 589:111–135
Bhattacharya S (2011) Natural antimutagens: a review. Res J Med Plants 5:116–126
Botelho RG, Christofoletti CA, Correia JE, Ansoar Y, Olinda RA, Tornisielo VL (2015) Genotoxic responses of juvenile tilapia (Oreochromis niloticus) exposed to florfenicol and oxytetracycline. Chemosphere 132:206–212
Braham RP, Blazer VS, Shaw CH, Mazik PM (2017) Micronuclei and other erythrocyte nuclear abnormalities in fishes from the Great Lakes Basin, USA. Environ Mol Mutagen 58:570–581
Carrasco K, Tilbury K, Myers M (1990) Assessment of the piscine micronuclei test as an in situ biological indicator of chemical contaminant effects. Can J Fish Aquat Sci 47:2123–2136
Castro D, Mieiro CL, Coelho JP, Guilherme S, Marques A, Santos MA, Duarte AC, Pereira E, Pacheco M (2018) Addressing the impact of mercury estuarine contamination in the European eel (Anguilla anguilla L., 1758) – An early diagnosis in glass eel stage based on erythrocytic nuclear morphology. Mar Pollut Bull 127:733–742
Cavas T, Garanko NN, Arkhipchuk VV (2005) Induction of micronuclei and binuclei in blood, gill and liver cells of fishes subchronically exposed to cadmium chloride and copper sulphate. Food Chem Toxicol 43:569–574
Celikler S, Tas S, Vatan O, Ziyanok-Ayvalik S, Yildiz G, Bilaloglu R (2009a) Anti-hyperglycemic and antigenotoxic potential of Ulva rigida ethanolic extract in the experimental diabetes mellitus. Food Chem Toxicol 47:1837–1840
Celikler S, Tas S, Ziyanok-Ayvalik S, Vatan O, Yildiz G, Ozel M (2014) Protective and antigenotoxic effect of Ulva rigida C. Agardh in experimental hypothyroid. Acta Biol Hung 65:13–26
Celikler S, Vatan O, Yildiz G, Bilaloglu R (2009b) Evaluation of anti-oxidative, genotoxic and antigenotoxic potency of Codium tomentosum Stackhouse ethanolic extract in human lymphocytes in vitro. Food Chem Toxicol 47:796–801
Costa S, Coelho P, Costa C, Silva S, Mayan O, Santos LS, Gaspar J, Teixeira JP (2008) Genotoxic damage in pathology anatomy laboratory workers exposed to formaldehyde. Toxicology 252:40–48
Craft TR, Bermudez E, Skopek TR (1987) Formaldehyde mutagenesis and formation of DNA-protein crosslinks in human lymphoblasts in vitro. Mutat Res Mol Mech Mutagen 176:147–155
Dantagnan P, Hernández A, Borquez A, Mansilla A (2009) Inclusion of macroalgae meal (Macrocystis pyrifera) as feed ingredient for rainbow trout (Oncorhynchus mykiss): effect on flesh fatty acid composition. Aquac Res 41:87–94
Díaz-Rubio ME, Pérez-Jiménez J, Saura-Calixto F (2009) Dietary fiber and antioxidant capacity in Fucus vesiculosus products. Int J Food Sci Nutr 60:23–34
DrugBank (2005) Cyclophosphamide. https://www.drugbank.ca/drugs/DB00531.
Ebrahimzadeh MA, Khalili M, Dehpour AA (2018) Antioxidant activity of ethyl acetate and methanolic extracts of two marine algae, Nannochloropsis oculata and Gracilaria gracilis - an in vitro assay. Brazilian J Pharm Sci. https://doi.org/10.1590/s2175-97902018000117280
El-Sayed A-KIM, Soltan MA, Radwan HA, Mohamed MG (2013) Effect of oxytetracycline and florfenicol on the cytogenetic picture of Nile tilapia (Oreochromis niloticus) fish. J Appl Biol Sci 7:102–106
Farvin KHS, Jacobsen C (2013) Phenolic compounds and antioxidant activities of selected species of seaweeds from Danish coast. Food Chem 138:1670–1681
Fenech M (2000) The in vitro micronucleus technique. Mutat Res 455:81–95
Fleurence J, Morançais M, Dumay J, Decottignies P, Turpin V, Munier M, Garcia-Bueno N, Jaouen P (2012) What are the prospects for using seaweed in human nutrition and for marine animals raised through aquaculture? Trends Food Sci Technol 27:57–61
Francavilla M, Franchi M, Monteleone M, Caroppo C (2013) The red seaweed Gracilaria gracilis as a multi products source. Mar Drugs 11:3754–3776
Gamal-Eldeen AM, Abo-Zeid MAM, Ahmed EF (2013) Anti-genotoxic effect of the Sargassum dentifolium extracts: prevention of chromosomal aberrations, micronuclei, and DNA fragmentation. Exp Toxicol Pathol 65:27–34
Garcia-Vaquero M, Hayes M (2016) Red and green macroalgae for fish and animal feed and human functional food development. Food Rev Int 32:15–45
Gomes JMM, Ribeiro HJ, Procópio MS, Alvarenga BM, Castro AC, Dutra WO, da Silva JB, Corrêa Junior JD (2015) What the erythrocytic nuclear alteration frequencies could tell us about genotoxicity and macrophage iron storage? PLoS One 10:1–22
Grafstrom RC, Curren RD, Yang LL, Harris CC (1985) Genotoxicity of formaldehyde in cultured human bronchial fibroblasts. Science 228:89–91
Grisolia CK (2002) A comparison between mouse and fish micronucleus test using cyclophosphamide, mitomycin C and various pesticides. Mutat Res 518:145–150
Guilherme S, Válega M, Pereira ME, Santos MA, Pacheco M (2008) Erythrocytic nuclear abnormalities in wild and caged fish (Liza aurata) along an environmental mercury contamination gradient. Ecotoxicol Environ Saf 70:411–421
Heffernan N, Smyth TJ, Soler-Villa A, Fitzgerald RJ, Brunton NP (2015) Phenolic content and antioxidant activity of fractions obtained from selected Irish macroalgae species (Laminaria digitata, Fucus serratus, Gracilaria gracilis and Codium fragile). J Appl Phycol 27:519–530
Henriques MAR (1998) Manual de aquacultura. Programa Operacional do Norte, Porto 207 pp
Higashi-Okai K, Otani S, Okai Y (1999) Potent suppressive effect of a Japanese edible seaweed, Enteromorpha prolifera (Sujiao-nori) on initiation and promotion phases of chemically induced mouse skin tumorigenesis. Cancer Lett 140:21–25
Holdt SL, Kraan S (2011) Bioactive compounds in seaweed: functional food applications and legislation. J Appl Phycol 23:543–597
IARC (2006) Formaldehyde, 2-butoxyethanol and 1-tert-butoxypropan-2-ol. IARC Monogr Eval Carcinog Risks Hum 88 478 pp
Izquierdo-Vega J, Morales-González J, Sánchez-Gutiérrez M, Betanzos-Cabrera G, Sosa-Delgado SM, Sumaya-Martínez MT, Morales-González A, Paniagua-Pérez R, Madrigal-Bujaidar E, Madrigal-Santillán E (2017) Evidence of some natural products with antigenotoxic effects. Part 1: fruits and polysaccharides. Nutrients 9:102
Jerbi MA, Ouanes Z, Besbes R, Achour L, Kacem A (2011) Single and combined genotoxic and cytotoxic effects of two xenobiotics widely used in intensive aquaculture. Mutat Res Toxicol Environ Mutagen 724:22–27
Jung SH, Kim JW, Jeon IG, Lee YH (2001) Formaldehyde residues in formalin-treated olive flounder (Paralichthys olivaceus), black rockfish (Sebastes schlegeli), and seawater. Aquaculture 194:253–262
Jung SH, Sim DS, Park M-S, Jo Q, Kim Y (2003) Effects of formalin on haematological and blood chemistry in olive flounder, Paralichthys olivaceus (Temminck et Schlegel). Aquac Res 34:1269–1275
Khan MA, Musarrat J (2003) Interactions of tetracycline and its derivatives with DNA in vitro in presence of metal ions. Int J Biol Macromol 33:49–56
Khan MA, Mustafa J, Musarrat J (2003) Mechanism of DNA strand breakage induced by photosensitized tetracycline–Cu(II) complex. Mutat Res Mol Mech Mutagen 525:109–119
Kiernan JA (2000) Formaldehyde, formalin, paraformaldehyde and glutaraldehyde: what they are and what they do. Micros Today 8:8–13
Koury MJ, Ponka P (2004) New insights into erythropoiesis: the roles of folate, vitamin B12, and iron. Annu Rev Nutr 24:105–131
Kreutzmann H-L (1977) The effects of chloramphenicol and oxytetracycline on haematopoiesis in the European eel (Anguilla anguilla). Aquaculture 10:323–334
Lahaye M, Robic A (2007) Structure and functional properties of ulvan, a polysaccharide from green seaweeds. Biomacromolecules 8:1765–1774
Leal JF, Neves MGPMS, Santos EBH, Esteves VI (2018) Use of formalin in intensive aquaculture: properties, application and effects on fish and water quality. Rev Aquac 10:281–295
Ma T-H, Harris MM (1988) Review of the genotoxicity of formaldehyde. Mutat Res Genet Toxicol 196:37–59
Maceda-Veiga A, Figuerola J, Martínez-Silvestre A, Viscor G, Ferrari N, Pacheco M (2015) Inside the Redbox: applications of haematology in wildlife monitoring and ecosystem health assessment. Sci Total Environ 514:322–332
Maceda-Veiga A, Monroy M, Viscor G, De Sostoa A (2010) Changes in non-specific biomarkers in the Mediterranean barbel (Barbus meridionalis) exposed to sewage effluents in a Mediterranean stream (Catalonia, NE Spain). Aquat Toxicol 100:229–237
Marques A, Ferreira J, Abreu H, Pereira R, Rego A, Serôdio J, Christa G, Gaivão I, Pacheco M (2018) Searching for antigenotoxic properties of marine macroalgae dietary supplementation against endogenous and exogenous challenges. J Toxicol Environ Health A:1–18. https://doi.org/10.1080/15287394.2018.1507856
Mert R, Karasu Benli AÇ, Arslan G (2015) Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.). Aquac Res 46:2798–2807
Mezghani S, Csupor D, Bourguiba I, Hohmann J, Amri M, Bouaziz M (2016) Characterization of phenolic compounds of Ulva rigida (Chlorophycae) and its antioxidant activity. Eur J Med Plants 12:1–9
Mohamed S, Hashim SN, Rahman HA (2012) Seaweeds: a sustainable functional food for complementary and alternative therapy. Trends Food Sci Technol 23:83–96
Morel J-M, Perrey F, Lejeune R, Goetz P (2005) Fucus vesiculosus L. Phytothérapie 3:218–221
Nagarani N, JanakiDevi V, YokeshBabu M, Kumaraguru AK (2012) Protective effect of Kappaphycus alvarezii (Rhodophyta) extract against DNA damage induced by mercury chloride in marine fish. Toxicol Environ Chem 94:1401–1410
Okai Y, Higashi-Okai K (1994) Identification of antimutagenic activities in the extract of an edible brown alga, Hijikia fusiforme (Hijiki) by umu gene expression system in Salmonella typhimurium (TA 1535/pSK 1002). J Sci Food Agric 66:103–109
Olson MH, Mitchell DL (2006) lnterspecific variation in UV defense mechanisms among temperate freshwater fishes. Photochem Photobiol 82:606–610
Omoregie E, Oyebanji SM (2002) Oxytetracycline-induced blood disorder in juvenile Nile tilapia Oreochromis niloticus (Trewavas). J World Aquacult Soc 33:377–382
Pacheco M, Santos MA (1998) Induction of liver EROD and erythrocytic nuclear abnormalities by cyclophosphamide and PAHs in Anguilla anguilla L. Ecotoxicol Environ Saf 40:71–76
Pacheco M, Santos MA (2002) Biotransformation, genotoxic, and histopathological effects of environmental contaminants in European eel (Anguilla anguilla L.). Ecotoxicol Environ Saf 53:331–347
Pacheco M, Santos MA (1997) Induction of EROD activity and genotoxic effects by polycyclic aromatic hydrocarbons and resin acids on the juvenile eel (Anguilla anguilla L.). Ecotoxicol Environ Saf 38:252–259
Pacheco M, Santos MA (1996) Induction of micronuclei and nuclear abnormalities in the erythrocytes of Anguilla anguilla L. exposed either to cyclophosphamide or to bleached kraft pulp mill effluent. Fresenius Environ Bull 5:746–751
Pedersen L-F, Pedersen PB, Nielsen JL, Nielsen PH (2010) Long term/low dose formalin exposure to small-scale recirculation aquaculture systems. Aquac Eng 42:1–7
Peixoto MJ, Salas-Leitón E, Pereira LF, Queiroz A, Magalhães F, Pereira R, Abreu H, Reis PA, Gonçalves JFM, Ozório ROA (2016) Role of dietary seaweed supplementation on growth performance, digestive capacity and immune and stress responsiveness in European seabass (Dicentrarchus labrax). Aquac Reports 3:189–197
Pereira V, Marques A, Gaivão I, Rego A, Abreu H, Pereira R, Santos MA, Guilherme S, Pacheco M (2019) Marine macroalgae as a dietary source of genoprotection in gilthead seabream (Sparus aurata) against endogenous and exogenous challenges. Comp Biochem Physiol C. https://doi.org/10.1016/j.cbpc.2019.01.006
Pereira VSAS (2016) Protective effects of seaweed feed supplementation towards genetic integrity in gilthead seabream (Sparus aurata). Master thesis, http://hdl.handle.net/10773/22364
Queiroz AC, Pereira R, Domingues AF, Peixoto MJD, Gonçalves JFM, Ozorio ROA (2014) Effect of seaweed supplementation on growth performance, immune and oxidative stress responses in gilthead seabream (Sparus aurata). Front Mar Sci Conf Abstr Int Meet Mar Res
Rasyid A, Ardiansyah A, Pangestuti R (2019) Nutrient composition of dried seaweed Gracilaria gracilis. Indones J Mar Sci 24:1–6
Rigos G, Nengas I, Alexis M (2006) Oxytetracycline (OTC) uptake following bath treatment in gilthead sea bream (Sparus aurata). Aquaculture 261:1151–1155
Rigos G, Nengas I, Tyrpenou AE, Alexis M, Troisi GM (2003) Pharmacokinetics and bioavailability of oxytetracycline in gilthead sea bream (Sparus aurata) after a single dose. Aquaculture 221:75–83
Rodrigues S, Antunes SC, Correia AT, Nunes B (2017) Rainbow trout (Oncorhynchus mykiss) pro-oxidant and genotoxic responses following acute and chronic exposure to the antibiotic oxytetracycline. Ecotoxicology 26:104–117
Romero J, Feijoó CG, Navarrete P (2012) Antibiotics in aquaculture – use, abuse and alternatives. In: Carvalho E (ed) Health and environment in aquaculture. InTech, Riejeka pp 159–198
Rupérez P, Ahrazem O, Leal JA (2002) Potential antioxidant capacity of sulfated polysaccharides from the edible marine brown seaweed Fucus vesiculosus. J Agric Food Chem 50:840–845
Savage JRK (1993) Update on target theory as applied to chromosomal aberrations. Environ Mol Mutagen 22:198–207
Schlenk D, Benson WH (2001) Target organ toxicity in marine and freshwater teleosts: organs. Taylor & Francis, London
Sharker MR, Sumi KR, Alam MJ, Rahman MM, Ferdous Z, Ali MM, Chaklader MR (2014) Drugs and chemicals used in aquaculture activities for fish health management in the coastal region of Bangladesh. Int J Life Sci Biotechnol Pharma Res 3:49
Silva J, Laranjeira A, Serradeiro R, Santos MA, Pacheco M (2011) Ozonated seawater induces genotoxicity and hematological alterations in turbot (Scophthalmus maximus) — Implications for management of recirculation aquaculture systems. Aquaculture 318:180–184
Smith CE, Piper RG (1972) Pathological effects in formalin-treated rainbow trout (Salmo gairdneri). J Fish Res Bd Canada 29:328–329
Stoiber T, Bonacker D, Böhm K, Bolt HM, Thier R, Degen GH, Unger E (2004) Disturbed microtubule function and induction of micronuclei by chelate complexes of mercury (II). Mutat Res 563:97–106
Stuart KR, Keller M, Drawbridge M (2010) Efficacy of formalin and povidone–iodine disinfection techniques on the eggs of three marine finfish species. Aquac Res 41:e838–e843
Swenberg JA, Moeller BC, Lu K, Rager JE, Fry RC, Starr TB (2012) Formaldehyde carcinogenicity research: 30 years and counting for mode of action, epidemiology, and cancer risk assessment. Toxicol Pathol 41:181–189
Taboada C, Millán R, Míguez I (2010) Composition, nutritional aspects and effect on serum parameters of marine algae Ulva rigida. J Sci Food Agric 90:445–449
Valente LMP, Araújo M, Batista S, Peixoto MJ, Sousa-Pinto I, Brotas V, Cunha LM, Rema P (2016) Carotenoid deposition, flesh quality and immunological response of Nile tilapia fed increasing levels of IMTA-cultivated Ulva spp. J Appl Phycol 28:691–701
Valente LMP, Rema P, Ferraro V, Pintado M, Sousa-Pinto I, Cunha LM, Oliveira MB, Araújo M (2015) Iodine enrichment of rainbow trout flesh by dietary supplementation with the red seaweed Gracilaria vermiculophylla. Aquaculture 446:132–139
Wassef EA, El-Sayed AFM, Kandeel KM, Sakr EM (2005) Evaluation of Pterocladia (Rhodophyta) and Ulva (Chlorophyta) meals as additives to gilthead seabream Sparus aurata diets. Egypt J Aquat Res 31:321–332
Winter MJ, Ellis LCJ, Hutchinson TH (2007) Formation of micronuclei in erythrocytes of the fathead minnow (Pimephales promelas) after acute treatment with mitomycin C or cyclophosphamide. Mutat Res Toxicol Environ Mutagen 629:89–99
Wooster GA, Martinez CM, Bowser PR, O’Hara DS (2011) Human health risks associated with formalin treatments used in aquaculture: initial study. N Am J Aquac 67:111–113
Xie D, Li X, You C, Wang S, Li Y (2019) Supplementation of macroalgae together with non-starch polysaccharide-degrading enzymes in diets enhanced growth performance, innate immune indexes, and disease resistance against Vibrio parahaemolyticus in rabbitfish Siganus canaliculatus. J Appl Phycol 31:2073–2083
Yang JI, Yeh CC, Lee JC, Yi SC, Huang HW, Tseng CN, Chang HW (2012) Aqueous extracts of the edible Gracilaria tenuistipitata are protective against H2O2-induced DNA damage, growth inhibition, and cell cycle arrest. Molecules. https://doi.org/10.3390/molecules17067241
Yildiz G, Celikler S, Vatan O, Dere Ş (2012) Determination of the anti-oxidative capacity and bioactive compounds in green seaweed Ulva rigida C. Agardh. Int J Food Prop 15:1182–1189
Yildiz G, Dere E, Dere Ş (2014) Comparison of the antioxidative components of some marine macroalgae from Turkey. Pak J Bot 46:753–757
Yuan Y, Walsh N (2006) Antioxidant and antiproliferative activities of extracts from a variety of edible seaweeds. Food Chem Toxicol 44:1144–1150
Zar J (1996) Biostatistical analysis. Prentice Hall, New Jersey
Zinadah OA, Khalil WK, Ashmaoui HM, Abdu F, Alsoud MA (2013) Evaluation of the anti-genotoxicity and growth performance impacts of green algae on Mugil cephalus. Life Sci J 10:1543–1554
Zounková R, Klimešová Z, Nepejchalová L, Hilscherová K, Bláha L (2011) Complex evaluation of ecotoxicity and genotoxicity of antimicrobials oxytetracycline and flumequine used in aquaculture. Environ Toxicol Chem 30:1184–1189
Zubia M, Fabre MS, Kerjean V, Lann KL, Stiger-Pouvreau V, Fauchon M, Deslandes E (2009) Antioxidant and antitumoural activities of some Phaeophyta from Brittany coasts. Food Chem 116:693–701
Funding
Thanks are due for the financial support to CESAM (UID/AMB/50017 - POCI-01-0145-FEDER-007638), to FCT/MCTES through national funds (PIDDAC) and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020 and to FCT/MCTES through the Ph.D. fellowship (SFRH/BD/102671/2014). This work was also funded by the project UID/CVT/00772/2019 supported by FCT/MCTES.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interests
The authors declare that they have no conflicts of interest.
Ethics approval
This study was conducted in accordance with the EU Directive 2010/63/EU on the protection of animals used for scientific purposes, under the supervision of a certified operator authorized by the Portuguese Veterinary Directorate (approval no. 0421/000/000).
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(DOCX 450 kb)
Rights and permissions
About this article
Cite this article
Marques, A., Marçal, R., Pereira, V. et al. Macroalgae-enriched diet protects gilthead seabream (Sparus aurata) against erythrocyte population instability and chromosomal damage induced by aqua-medicines. J Appl Phycol 32, 1477–1493 (2020). https://doi.org/10.1007/s10811-019-01996-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-019-01996-2