Cytotoxicity of Mycotoxins Frequently Present in Aquafeeds to the Fish Cell Line RTGill-W1

In the last decades, the aquaculture industry has introduced plant-based ingredients as a source of protein in aquafeeds. This has led to mycotoxin contaminations, representing an ecological, health and economic problem. The aim of this study was to determine in the RTgill-W1 fish cell line the toxicity of fifteen mycotoxins of common occurrence in aquafeeds. To identify the most sensitive endpoint of toxicity, the triple assay was used. It consisted of three assays: alamarBlue, Neutral Red Uptake and CFDA-AM, which revealed the mitochondrial activity, the lysosomal integrity and the plasma membrane integrity, respectively. Most of the assayed mycotoxins were toxic predominantly at lysosomal level (enniatins, beauvericin, zearalenone, ochratoxin A, deoxynivalenol (DON) and its acetylated metabolites 15-O-acetyl-DON and 3-acetyl-DON). Aflatoxins B1 and B2 exerted the greatest effects at mitochondrial level, while fumonisins B1 and B2 and nivalenol were not toxic up to 100 µg/mL. In general, low toxicity was observed at plasma membrane level. The vast majority of the mycotoxins assayed exerted a pronounced acute effect in the fish RTgill-W1 cell line, emphasizing the need for further studies to ascertain the impact of mycotoxin contamination of fish feeds in the aquaculture industry and to establish safe limits in aquafeeds.

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Dynamics of Nontypical Apoptotic Morphological Changes Visualized by Green Fluorescent Protein in Living Cells with Infectious Pancreatic Necrosis Virus Infection

Journal of Virology ◽

10.1128/jvi.73.6.5056-5063.1999 ◽

1999 ◽

Vol 73 (6) ◽

pp. 5056-5063 ◽

Cited By ~ 30

Author(s):

Jiann-Ruey Hong ◽

Tai-Lang Lin ◽

Jer-Yen Yang ◽

Ya-Li Hsu ◽

Jen-Leih Wu

Keyword(s):

Cell Line ◽

Fluorescent Protein ◽

Morphological Changes ◽

Membrane Integrity ◽

Infectious Pancreatic Necrosis Virus ◽

Fish Cell Line ◽

Fish Cell ◽

Infectious Pancreatic Necrosis ◽

Pancreatic Necrosis Virus ◽

Green Fluorescent

ABSTRACT Morphologically, apoptotic cells are characterized by highly condensed membrane blebbing and formation of apoptotic bodies. Recently, we reported that apoptosis precedes necrosis in a fish cell line infected with infectious pancreatic necrosis virus (IPNV). In the present study, we tested the possibility that nontypical apoptosis is a component of IPNV-induced fish cell death. A variant type of green fluorescent protein (EGFP) was expressed in a fish cell line such that EGFP served as a protein marker for visualizing dynamic apoptotic cell morphological changes and for tracing membrane integrity changes during IPNV infection. Direct morphological changes were visualized by fluorescence microscopy by EGFP in living cells infected with IPNV. The nontypical apoptotic morphological change stage occurred during the pre-late stage (6 to 7 h postinfection). Nontypical apoptotic features, including highly condensed membrane blebbing, occurred during the middle apoptotic stage. At the pre-late apoptotic stage, membrane vesicles quickly formed, blebbed, and were finally pinched off from the cell membrane. At the same time, at this pre-late apoptotic stage, apoptotic cells formed unique small holes in their membranes that ranged from 0.39 to 0.78 μm according to examination by scanning electron microscopy and immunoelectron microscopy. Quantitation of the intra- and extracellular release of EGFP by CHSE-214-EGFP cells after IPNV infection was done by Western blotting and fluorometry. Membrane integrity was quickly lost during the late apoptotic stage (after 8 h postinfection), and morphological change and membrane integrity loss could be prevented and blocked by treatment with apoptosis inhibitors such as cycloheximide, genistein, and EDTA before IPNV infection. Together, these findings show the apoptotic features at the onset of pathology in host cells (early and middle apoptotic stages), followed secondarily by nontypical apoptosis (pre-late apoptotic stage) and then by postapoptotic necrosis (late apoptotic stage), of a fish cell line. Our results demonstrate that nontypical apoptosis is a component of IPNV-induced fish cell death.

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Growth of the penaeid shrimp virus infectious hypodermal and hematopoietic necrosis virus in a fish cell line

Journal of Virological Methods ◽

10.1016/0166-0934(90)90120-5 ◽

1990 ◽

Vol 28 (3) ◽

pp. 273-280 ◽

Cited By ~ 5

Author(s):

Philip C. Loh ◽

Yuanan Lu ◽

James A. Brock

Keyword(s):

Cell Line ◽

Penaeid Shrimp ◽

Fish Cell Line ◽

Fish Cell ◽

Necrosis Virus ◽

Shrimp Virus

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Plasma membrane integrity: implications for health and disease

BMC Biology ◽

10.1186/s12915-021-00972-y ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Dustin A. Ammendolia ◽

William M. Bement ◽

John H. Brumell

Keyword(s):

Plasma Membrane ◽

Membrane Damage ◽

Membrane Integrity ◽

Whole Body ◽

Plasma Membrane Integrity ◽

Plasma Membrane Damage ◽

Cellular Environment ◽

Health And Disease ◽

Repair Pathways ◽

The Impact

AbstractPlasma membrane integrity is essential for cellular homeostasis. In vivo, cells experience plasma membrane damage from a multitude of stressors in the extra- and intra-cellular environment. To avoid lethal consequences, cells are equipped with repair pathways to restore membrane integrity. Here, we assess plasma membrane damage and repair from a whole-body perspective. We highlight the role of tissue-specific stressors in health and disease and examine membrane repair pathways across diverse cell types. Furthermore, we outline the impact of genetic and environmental factors on plasma membrane integrity and how these contribute to disease pathogenesis in different tissues.

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