Development of Molecular and Histological Methods to Evaluate Stress Oxidative Biomarkers in Sea Bass (Dicentrarchus Labrax)

Overview

Journal Fish Physiol Biochem

Specialty Biochemistry

Date 2020 May 11

PMID 32388830

Citations 6

Authors

Eleonora Fiocchi

Michele Civettini

Pierluigi Carbonara

Walter Zupa

Giuseppe Lembo

Amedeo Manfrin

Affiliations

Soon will be listed here.

Abstract

In aquaculture, fish species may experience stressful episodes caused by poor farming conditions. The exponential increase of global aquaculture has raised the number of research studies aimed at demonstrating the sensitivity of aquatic animals in confined environments. The development of a real-time PCR and immunohistochemistry methods were investigated to evaluate the presence, localization, and quantity of biomarkers of oxidative stress in European sea bass (Dicentrarchus labrax). In particular, stress tests such as manipulation and temperature changes were conducted through molecular methods to identify the expression level of heat shock protein 70 (HSP70) in stressed animals compared with a control group. The immunohistochemical technique was also applied to locate and study the trends-levels of nitrotyrosine (NT), heat shock protein 70 (HSP70), malondialdehyde (MDA), and 4-hydroxy-2-nonenal (HNE) in different tissues from stressed animals and control group. The presence of the rodlet cell (RCs) was evaluated by histology in both a control and stressed group. Our results show that the real-time PCR method developed is specific for the evaluated target gene and that manipulation and temperature increase are strong stressors for animals. Relative quantification data revealed a gene expression increase of HSP70 in the stressed group of animals compared to the control group. The antibodies used for the immunohistochemical staining were efficient, and it was possible to appreciate the increase of immunoprecipitates in European sea bass either manipulated or stressed by temperature increase. The present study can be a starting point to allow the quantification of HSP70 and the identification of other stress biomarkers in D. labrax.

Citing Articles

Growth performance, gut microbiota composition, health and welfare of European sea bass (Dicentrarchus labrax) fed an environmentally and economically sustainable low marine protein diet in sea cages.

Alfonso S, Mente E, Fiocchi E, Manfrin A, Dimitroglou A, Papaharisis L Sci Rep. 2023; 13(1):21269.

PMID: 38042956 PMC: 10693626. DOI: 10.1038/s41598-023-48533-3.

Influence of Transportation on Stress Response and Cellular Oxidative Stress Markers in Juvenile Meagre ().

Bortoletti M, Fonsatti E, Leva F, Maccatrozzo L, Ballarin C, Radaelli G Animals (Basel). 2023; 13(20).

PMID: 37894012 PMC: 10603926. DOI: 10.3390/ani13203288.

Histological and Chemical Analysis of Heavy Metals in Kidney and Gills of : Melanomacrophages Centers and Rodlet Cells as Environmental Biomarkers.

Alesci A, Cicero N, Fumia A, Petrarca C, Mangifesta R, Nava V Toxics. 2022; 10(5).

PMID: 35622632 PMC: 9147125. DOI: 10.3390/toxics10050218.

Muscle Cortisol Levels, Expression of Glucocorticoid Receptor and Oxidative Stress Markers in the Teleost Fish Exposed to Transport Stress.

Bortoletti M, Maccatrozzo L, Radaelli G, Caberlotto S, Bertotto D Animals (Basel). 2021; 11(4).

PMID: 33919515 PMC: 8072758. DOI: 10.3390/ani11041160.

Salinity, Temperature and Ammonia Acute Stress Response in Seabream () Juveniles: A Multidisciplinary Study.

Zarantoniello M, Bortoletti M, Olivotto I, Ratti S, Poltronieri C, Negrato E Animals (Basel). 2021; 11(1).

PMID: 33419050 PMC: 7825456. DOI: 10.3390/ani11010097.

References

Aldini G, Dalle-Donne I, Maffei Facino R, Milzani A, Carini M . Intervention strategies to inhibit protein carbonylation by lipoxidation-derived reactive carbonyls. Med Res Rev. 2006; 27(6):817-68. DOI: 10.1002/med.20073. View

Boscolo Papo M, Bertotto D, Quaglio F, Vascellari M, Pascoli F, Negrato E . Histopathology and stress biomarkers in the clam Venerupis philippinarum from the Venice Lagoon (Italy). Fish Shellfish Immunol. 2014; 39(1):42-50. DOI: 10.1016/j.fsi.2014.04.016. View

Dezfuli B, Simoni E, Giari L, Manera M . Effects of experimental terbuthylazine exposure on the cells of Dicentrarchus labrax (L.). Chemosphere. 2006; 64(10):1684-94. DOI: 10.1016/j.chemosphere.2006.01.023. View

Giari L, Simoni E, Manera M, Dezfuli B . Histo-cytological responses of Dicentrarchus labrax (L.) following mercury exposure. Ecotoxicol Environ Saf. 2007; 70(3):400-10. DOI: 10.1016/j.ecoenv.2007.08.013. View

Gornati R, Papis E, Rimoldi S, Terova G, Saroglia M, Bernardini G . Rearing density influences the expression of stress-related genes in sea bass (Dicentrarchus labrax, L.). Gene. 2004; 341:111-8. DOI: 10.1016/j.gene.2004.06.020. View

Iger Y, Abraham M . Rodlet cells in the epidermis of fish exposed to stressors. Tissue Cell. 1997; 29(4):431-8. DOI: 10.1016/s0040-8166(97)80029-8. View

Ischiropoulos H, Zhu L, Chen J, Tsai M, Martin J, Smith C . Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase. Arch Biochem Biophys. 1992; 298(2):431-7. DOI: 10.1016/0003-9861(92)90431-u. View

Mosca F, Ciulli S, Volpatti D, Romano N, Volpe E, Bulfon C . Defensive response of European sea bass (Dicentrarchus labrax) against Listonella anguillarum or Photobacterium damselae subsp. piscicida experimental infection. Vet Immunol Immunopathol. 2014; 162(3-4):83-95. DOI: 10.1016/j.vetimm.2014.10.002. View

Pascoli F, Negrato E, Di Giancamillo A, Bertotto D, Domeneghini C, Simontacchi C . Evaluation of oxidative stress biomarkers in Zosterisessor ophiocephalus from the Venice Lagoon, Italy. Aquat Toxicol. 2011; 101(3-4):512-20. DOI: 10.1016/j.aquatox.2010.12.003. View

10.

Patruno M, Maccatrozzo L, Funkenstein B, Radaelli G . Cloning and expression of insulin-like growth factors I and II in the shi drum (Umbrina cirrosa). Comp Biochem Physiol B Biochem Mol Biol. 2006; 144(2):137-51. DOI: 10.1016/j.cbpb.2006.02.003. View

11.

Poltronieri C, Maccatrozzo L, Simontacchi C, Bertotto D, Funkenstein B, Patruno M . Quantitative RT-PCR analysis and immunohistochemical localization of HSP70 in sea bass Dicentrarchus labrax exposed to transport stress. Eur J Histochem. 2007; 51(2):125-35. View

12.

Reite O . The rodlet cells of teleostean fish: their potential role in host defence in relation to the role of mast cells/eosinophilic granule cells. Fish Shellfish Immunol. 2005; 19(3):253-67. DOI: 10.1016/j.fsi.2005.01.002. View

13.

Reite O, Evensen O . Inflammatory cells of teleostean fish: a review focusing on mast cells/eosinophilic granule cells and rodlet cells. Fish Shellfish Immunol. 2005; 20(2):192-208. DOI: 10.1016/j.fsi.2005.01.012. View