The increasing antibiotic resistance has led to a search for alternative antimicrobial agents. Natural deep eutectic solvents (NADESs) have emerged as promising alternatives due to their low volatility, low flammability, ease of preparation, and low cost. This review discusses the antimicrobial properties of multiple NADESs, focusing on their antibacterial and antifungal activities. NADESs are prepared using various methods, including heating and stirring, freeze-drying, evaporation, grinding, and ultrasound-assisted and microwave-assisted synthesis. The antimicrobial activity of NADESs is evaluated using techniques such as the disc diffusion method, broth dilution, Microtox assay, drop plate, and FTIR-based bioassays. The review highlights the strong antibacterial activity of NADESs against both gram-positive and gram-negative bacteria, with organic acids-based NADESs showing the highest toxicity. Additionally, NADESs exhibit significant antifungal activity against *Candida albicans*. The mechanisms of action of NADESs against bacteria and fungi are also discussed, including the role of hydrogen bond donors and acceptors. Overall, NADESs show great potential as alternatives to traditional antibiotics, particularly in addressing the challenge of antibiotic resistance.The increasing antibiotic resistance has led to a search for alternative antimicrobial agents. Natural deep eutectic solvents (NADESs) have emerged as promising alternatives due to their low volatility, low flammability, ease of preparation, and low cost. This review discusses the antimicrobial properties of multiple NADESs, focusing on their antibacterial and antifungal activities. NADESs are prepared using various methods, including heating and stirring, freeze-drying, evaporation, grinding, and ultrasound-assisted and microwave-assisted synthesis. The antimicrobial activity of NADESs is evaluated using techniques such as the disc diffusion method, broth dilution, Microtox assay, drop plate, and FTIR-based bioassays. The review highlights the strong antibacterial activity of NADESs against both gram-positive and gram-negative bacteria, with organic acids-based NADESs showing the highest toxicity. Additionally, NADESs exhibit significant antifungal activity against *Candida albicans*. The mechanisms of action of NADESs against bacteria and fungi are also discussed, including the role of hydrogen bond donors and acceptors. Overall, NADESs show great potential as alternatives to traditional antibiotics, particularly in addressing the challenge of antibiotic resistance.