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Potential (co-)contamination of dairy milk with AFM1 and MC-LR and their synergistic interaction in inducing mitochondrial dysfunction in HepG2 cells

Mohamed Fathi Abdallah (UGent) , Jessa May Recote, Camille Van Camp, Wannes H.R. Van Hasse, Lorenzo Pedroni, Luca Dellafiora, Julien Masquelier and Andreja Rajkovic (UGent)
(2024) FOOD AND CHEMICAL TOXICOLOGY. 192.
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Abstract
Several toxic metabolites, such as aflatoxin M1 (AFM1), are known to contaminate dairy milk. However, as mentioned in an external EFSA report, there is a knowledge gap regarding the carry-over of certain emerging toxins such as microcystin-LR (MC-LR). Therefore, this work aimed to develop an LC-MS/MS method for MC-LR quantification in dairy milk. Also, the method included AFM1 as a common fungal metabolite and applied to analyze 113 dairy milk samples collected directly after the end of the summer peak. Both toxins were below their LODs, keeping the question on MC-LR carry-over still unanswered. Moreover, an in silico analysis, using a 3D molecular modeling was performed, pointing to a possible interaction between MC-LR and milk proteins, especially β-lactoglobulin. Since AFM1 and MC-LR are hepatotoxic, their interaction in inducing mitochondrial dysfunction in HepG2 cells was investigated at low (subcytotoxic) concentrations. Live cell imaging-based assays showed an inhibition in cell viability, without involvement of caspase-3/7, and a hyperpolarization in the mitochondrial membrane potential after the exposure to a mixture of 100 ng mL-1 AFM1 and 1000 ng mL-1 MC-LR for 48h. Extracellular flux analysis revealed inhibitions of several key parameters of mitochondrial function (basal respiration, ATP-linked respiration, and spare respiratory capacity).
Keywords
Aflatoxin M1, Microcystin-LR, LC-MS/MS, Mitochondrial Toxicity, Live-cell Imaging, Oxygen Consumption Rate, HepG2, in silico 3D Modelling, PERMEABILITY TRANSITION, CYANOBACTERIAL TOXINS, PHYTOTOXICITY, MICROCYSTINS, APOPTOSIS

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Citation

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MLA
Abdallah, Mohamed Fathi, et al. “Potential (Co-)Contamination of Dairy Milk with AFM1 and MC-LR and Their Synergistic Interaction in Inducing Mitochondrial Dysfunction in HepG2 Cells.” FOOD AND CHEMICAL TOXICOLOGY, vol. 192, 2024, doi:10.1016/j.fct.2024.114907.
APA
Abdallah, M. F., Recote, J. M., Van Camp, C., Van Hasse, W. H. R., Pedroni, L., Dellafiora, L., … Rajkovic, A. (2024). Potential (co-)contamination of dairy milk with AFM1 and MC-LR and their synergistic interaction in inducing mitochondrial dysfunction in HepG2 cells. FOOD AND CHEMICAL TOXICOLOGY, 192. https://doi.org/10.1016/j.fct.2024.114907
Chicago author-date
Abdallah, Mohamed Fathi, Jessa May Recote, Camille Van Camp, Wannes H.R. Van Hasse, Lorenzo Pedroni, Luca Dellafiora, Julien Masquelier, and Andreja Rajkovic. 2024. “Potential (Co-)Contamination of Dairy Milk with AFM1 and MC-LR and Their Synergistic Interaction in Inducing Mitochondrial Dysfunction in HepG2 Cells.” FOOD AND CHEMICAL TOXICOLOGY 192. https://doi.org/10.1016/j.fct.2024.114907.
Chicago author-date (all authors)
Abdallah, Mohamed Fathi, Jessa May Recote, Camille Van Camp, Wannes H.R. Van Hasse, Lorenzo Pedroni, Luca Dellafiora, Julien Masquelier, and Andreja Rajkovic. 2024. “Potential (Co-)Contamination of Dairy Milk with AFM1 and MC-LR and Their Synergistic Interaction in Inducing Mitochondrial Dysfunction in HepG2 Cells.” FOOD AND CHEMICAL TOXICOLOGY 192. doi:10.1016/j.fct.2024.114907.
Vancouver
1.
Abdallah MF, Recote JM, Van Camp C, Van Hasse WHR, Pedroni L, Dellafiora L, et al. Potential (co-)contamination of dairy milk with AFM1 and MC-LR and their synergistic interaction in inducing mitochondrial dysfunction in HepG2 cells. FOOD AND CHEMICAL TOXICOLOGY. 2024;192.
IEEE
[1]
M. F. Abdallah et al., “Potential (co-)contamination of dairy milk with AFM1 and MC-LR and their synergistic interaction in inducing mitochondrial dysfunction in HepG2 cells,” FOOD AND CHEMICAL TOXICOLOGY, vol. 192, 2024.
@article{01J4KBW0YYV8YK4YQ5XX3PS0KH,
  abstract     = {{Several toxic metabolites, such as aflatoxin M1 (AFM1), are known to contaminate dairy milk. However, as mentioned in an external EFSA report, there is a knowledge gap regarding the carry-over of certain emerging toxins such as microcystin-LR (MC-LR). Therefore, this work aimed to develop an LC-MS/MS method for MC-LR quantification in dairy milk. Also, the method included AFM1 as a common fungal metabolite and applied to analyze 113 dairy milk samples collected directly after the end of the summer peak. Both toxins were below their LODs, keeping the question on MC-LR carry-over still unanswered. Moreover, an in silico analysis, using a 3D molecular modeling was performed, pointing to a possible interaction between MC-LR and milk proteins, especially β-lactoglobulin.

Since AFM1 and MC-LR are hepatotoxic, their interaction in inducing mitochondrial dysfunction in HepG2 cells was investigated at low (subcytotoxic) concentrations. Live cell imaging-based assays showed an inhibition in cell viability, without involvement of caspase-3/7, and a hyperpolarization in the mitochondrial membrane potential after the exposure to a mixture of 100 ng mL-1 AFM1 and 1000 ng mL-1 MC-LR for 48h. Extracellular flux analysis revealed inhibitions of several key parameters of mitochondrial function (basal respiration, ATP-linked respiration, and spare respiratory capacity).}},
  articleno    = {{114907}},
  author       = {{Abdallah, Mohamed Fathi and Recote, Jessa May and Van Camp, Camille and Van Hasse, Wannes H.R. and Pedroni, Lorenzo and Dellafiora, Luca and Masquelier, Julien and Rajkovic, Andreja}},
  issn         = {{0278-6915}},
  journal      = {{FOOD AND CHEMICAL TOXICOLOGY}},
  keywords     = {{Aflatoxin M1,Microcystin-LR,LC-MS/MS,Mitochondrial Toxicity,Live-cell Imaging,Oxygen Consumption Rate,HepG2,in silico 3D Modelling,PERMEABILITY TRANSITION,CYANOBACTERIAL TOXINS,PHYTOTOXICITY,MICROCYSTINS,APOPTOSIS}},
  language     = {{eng}},
  pages        = {{13}},
  title        = {{Potential (co-)contamination of dairy milk with AFM1 and MC-LR and their synergistic interaction in inducing mitochondrial dysfunction in HepG2 cells}},
  url          = {{http://doi.org/10.1016/j.fct.2024.114907}},
  volume       = {{192}},
  year         = {{2024}},
}
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