Experimental evidence of the anti-bacterial activity pathway of copper ion treatment on Mycobacterium avium subsp. paratuberculosis
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Updated: Aug 16, 2025
Studying Copper Nanoparticle-Induced Programmed Cell Death in Bacteria
Published on: May 16, 2025
Carlos Tejeda1,2, Marcela Villegas1, Pamela Steuer1
1Instituto de Medicina Preventiva Veterinaria, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Valdivia, Chile.
Abstract:
Copper causes significant damage to the integrity of many bacteria, mainly at the DNA level, through its redox states, as well as its reactive oxygen species (ROS) generating capacity at the cellular level. But whether these mechanisms also apply to Mycobacterium avium subsp. paratuberculosis (MAP) is unknown. In the present study, we have evaluated whether copper ions produce damage at the DNA level of MAP, either through their redox states or through ROS production. MAP-spiked PBS was first supplemented with different copper chelators (2) and ROS antioxidants (3), followed by treatment with copper ions at 942 ppm. MAP DNA integrity (qPCR, magnetic phage separation) was then evaluated. We found that bathocuproine (BCS), as a chelator, and D-mannitol, as an antioxidant of hydroxyl radicals, had a significant protective effect (P < 0.05) on DNA molecules, and that EDTA, as a chelator, and D-mannitol, as an antioxidant had a significant positive effect (P < 0.05) on the viability of this pathogen in contrast to the control and other chelators and anti-oxidants used. In light of the reported findings, it may be concluded that copper ions within MAP cells are directly related to MAP DNA damage.
Insights
Copper ions damage bacterial DNA. This study found copper causes DNA damage in Mycobacterium avium subsp. paratuberculosis (MAP) via redox states and reactive oxygen species (ROS).
Area of Science:
- Microbiology
- Environmental Science
- Biochemistry
Background:
- Copper is known to damage bacterial DNA through redox cycling and reactive oxygen species (ROS) generation.
- The specific mechanisms of copper-induced DNA damage in Mycobacterium avium subsp. paratuberculosis (MAP) remain largely uncharacterized.
Purpose of the Study:
- To investigate whether copper ions induce DNA damage in MAP.
- To determine if copper's DNA damaging effects are mediated by its redox states or ROS production.
Main Methods:
- MAP bacteria were exposed to copper ions (942 ppm) in phosphate-buffered saline.
- Experiments involved copper chelators (bathocuproine, EDTA) and ROS antioxidants (D-mannitol).
- MAP DNA integrity and viability were assessed using qPCR and magnetic phage separation.
Main Results:
- Bathocuproine (chelator) and D-mannitol (antioxidant) significantly protected MAP DNA molecules from copper-induced damage (P < 0.05).
- EDTA (chelator) and D-mannitol also significantly enhanced MAP viability.
- These protective effects were observed in contrast to control groups and other tested chelators/antioxidants.
Conclusions:
- Copper ions are directly implicated in causing DNA damage within MAP cells.
- The findings suggest copper's detrimental effects on MAP involve both direct DNA interaction and ROS generation.
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