Effect of chloride concentration on the cytotoxicity, bioavailability, and bioreactivity of copper and silver in the rainbow trout gut cell line, RTgutGC

Ibrahim, Md.; Minghetti, Matteo

doi:10.1007/s10646-022-02543-5

Cited by 3 publications

(13 citation statements)

References 43 publications

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“…Exposure regime 3 had the highest Ag-to-Cu ratio, which resulted in the highest empirically determined EC50 values for the mixture deviating from the CA prediction the most and suggesting that a high Ag-to-Cu ratio may result in antagonism and less predictability from the CA model. This effect is unexpected because previously it was shown that Ag is approximately three times more toxic than Cu in RTgutGC cells (Ibrahim & Minghetti, 2022;Ibrahim et al, 2020). These results suggest that free ionic species of Ag and Cu compete for cellular entry FIGURE 3: Median effective concentration (EC50) of the Ag-Cu mixture in toxic units obtained using the concentration addition design.…”

Section: Discussionmentioning

confidence: 92%

“…Two synthetic exposure media were prepared as previously described (Ibrahim & Minghetti, 2022) based on the composition of commercial L-15 medium but with different chloride (Cl − ) concentration (Supporting Information, Table S1). The concentration of Clwas 1 mM in L-15/ex 1Cland 146 mM in L-15/ ex 146Cl-; L-15/ex 1Cland L-15/ex 146Clhave similar sugar and salt concentrations as of L-15, but they lack amino acids and vitamins to avoid aminoacidic complexation with metals (Ibrahim & Minghetti, 2022). The osmolality and the pH of both media were measured using the Vapro ® Vapor Pressure Osmometer (Model 5600; ELItech Group, South Logan, UT) and an SI Analytics pH meter (SI Analytics, College Station, TX), respectively.…”

Section: Exposurementioning

confidence: 99%

“…The second design evaluated the predictability of the CA model. The EC50 of single-metal exposure, based on previous data (Ibrahim & Minghetti, 2022), was considered as 1 TU, and the range of summed TUs varied from 0.31 to 3.28 TUs. The exposure concentrations and TUs of Ag and Cu in micromolars are reported in Table 1.…”

Section: Exposurementioning

confidence: 99%

“…In low-Clmedium, where Ag and Cu are present in solution as free metal ions at similar concentrations, Cu bioavailability is not affected by the presence of Ag; but in high-Clmedium, Cu bioavailability is increased by fourfold. Conversely, Ag Ibrahim and Minghetti (2022). Statistical significance between expected and measured cell viability was determined using a Student t test (p < 0.05), and significance is indicated by asterisks.…”

Section: Bioavailability Of the Ag And Cu Mixture In Rtgutgc Cells Ex...mentioning

confidence: 99%

“…In the CA design, we determined combined EC50 in TUs of Ag and Cu coexposure by considering the individual EC50s of Ag and Cu as one TU while exposing cells to the mixture of Ag and Cu. The EC50s for single-metal exposure (i.e., Ag or Cu alone) were determined in a previous study, using the same cell line (Ibrahim & Minghetti, 2022). In addition, we were interested in explaining how metal speciation in the exposure medium affects the toxicity and bioavailability of Ag and Cu exposed in mixture to RTgutGC cells.…”

Section: Introductionmentioning

confidence: 99%

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Interactive Effects of Copper–Silver Mixtures at the Intestinal Epithelium of Rainbow Trout: An In Vitro Approach

Ibrahim,

Belden,

Minghetti

2023

Enviro Toxic and Chemistry

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While metals are present in mixture in the environment, metal toxicity studies are usually conducted on an individual metal basis. There is a paucity of data in the existing literature regarding specific metal‐metal interactions and their effect on metal toxicity and bioavailability. Here, we studied interactions of silver‐copper mixture at the intestinal epithelium using an intestinal cell line derived from rainbow trout (Oncorhynchus mykiss), the RTgutGC. Exposures were conducted in media containing different chloride concentrations (low chloride, 1 mM and high chloride, 146 mM) thus resulting in different metal speciation. Cytotoxicity was evaluated based on two endpoints, cell metabolic activity and cell membrane integrity. Silver‐copper mixture toxicity was assessed using two designs: the independent action (IA) and concentration addition (CA). Metal mixture bioavailability was studied by exposing cells to 500 nM of Ag or Cu in single or mixture exposure (i.e., 500 nM of Cu plus 500 nM of Ag). We found an antagonistic effect in the low chloride medium and an additive/synergistic effect in the high chloride medium. We found that copper (Cu) dominates over silver (Ag) toxicity and bioavailability indicating a competitive inhibition when both metals are present as free metal ions in the exposure media which support our hypothesis. Our study also suggests different mechanisms of uptake of free metal ions and metal complexes. This study adds valuable information to our understanding of the role of metal speciation on metal mixture toxicity and bioavailability.

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Section: Discussionmentioning

confidence: 92%

Section: Exposurementioning

confidence: 99%

Section: Exposurementioning

confidence: 99%

Section: Bioavailability Of the Ag And Cu Mixture In Rtgutgc Cells Ex...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interactive Effects of Copper–Silver Mixtures at the Intestinal Epithelium of Rainbow Trout: An In Vitro Approach

Ibrahim,

Belden,

Minghetti

2023

Enviro Toxic and Chemistry

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Role of the luminal composition on intestinal metal toxicity, bioavailability and bioreactivity: An in vitro approach based on the cell line RTgutGC

et al. 2023

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Establishment of an In Vitro Model to Study Viral Infections of the Fish Intestinal Epithelium

Løkka

Gamil

Evensen

et al. 2023

Cells

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Viral infections are still a major concern for the aquaculture industry. For salmonid fish, even though breeding strategies and vaccine development have reduced disease outbreaks, viral diseases remain among the main challenges having a negative impact on the welfare of fish and causing massive economic losses for the industry. The main entry port for viruses into the fish is through mucosal surfaces including that of the gastrointestinal tract. The contradictory functions of this surface, both creating a barrier towards the external environment and at the same time being responsible for the uptake of nutrients and ion/water regulation make it particularly vulnerable. The connection between dietary components and viral infections in fish has been poorly investigated and until now, a fish intestinal in vitro model to investigate virus–host interactions has been lacking. Here, we established the permissiveness of the rainbow trout intestinal cell line RTgutGC towards the important salmonid viruses—infectious pancreatic necrosis virus (IPNV), salmonid alphavirus (subtype 3, SAV3) and infectious salmon anemia virus (ISAV)—and explored the infection mechanisms of the three different viruses in these cells at different virus to cell ratios. Cytopathic effect (CPE), virus replication in the RTgutGC cells, antiviral cell responses and viral effects on the barrier permeability of polarized cells were investigated. We found that all virus species infected and replicated in RTgutGC cells, although with different replication kinetics and ability to induce CPE and host responses. The onset and progression of CPE was more rapid at high multiplicity of infection (MOI) for IPNV and SAV3 while the opposite was true of ISAV. A positive correlation between the MOI used and the induction of antiviral responses was observed for IPNV while a negative correlation was detected for SAV3. Viral infections compromised barrier integrity at early time points prior to observations of CPE microscopically. Further, the replication of IPNV and ISAV had a more pronounced effect on barrier function than SAV3. The in vitro infection model established herein can thus provide a novel tool to generate knowledge about the infection pathways and mechanisms used to surpass the intestinal epithelium in salmonid fish, and to study how a virus can potentially compromise gut epithelial barrier functions.

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Effect of chloride concentration on the cytotoxicity, bioavailability, and bioreactivity of copper and silver in the rainbow trout gut cell line, RTgutGC

Cited by 3 publications

References 43 publications

Interactive Effects of Copper–Silver Mixtures at the Intestinal Epithelium of Rainbow Trout: An In Vitro Approach

Interactive Effects of Copper–Silver Mixtures at the Intestinal Epithelium of Rainbow Trout: An In Vitro Approach

Role of the luminal composition on intestinal metal toxicity, bioavailability and bioreactivity: An in vitro approach based on the cell line RTgutGC

Establishment of an In Vitro Model to Study Viral Infections of the Fish Intestinal Epithelium

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