Effectiveness of botanical treatments has been recognized by many, but scientific validation on the beneficial use of plants is scarce. Nopalea pads are probably native to Mexico or Central America and are used for human consumption and for medicinal purposes. In this study, in vitro antibacterial and antifungal activities of fresh and dried Nopalea cochenillifera (NC) were investigated. MICs of fresh NC hexanic, chloroformic and ethanolic fractions against C. albicans were 250, 250 and 3.9 µg mL 1 respectively; MICs of fresh NC against Salmonella enterica var. thyphimurium were 15.6, 62.5 and 3.9 µg mL 1 respectively; and MICs of fresh NC against Escherichia coli were 500, NA (no activity) and 3.9 µg mL 1 respectively. In addition, MICs of dried NC hexanic, chloroformic and ethanolic fractions against C. albicans were 31.2, 31.2 and 3.9 µg mL 1 respectively; MICs of dried NC against Salmonella enterica var. thyphimurium were 15.6, NA and 3.9 µg mL 1 respectively; and MICs of dried NC against Escherichia coli were NA, 62.5 µg mL 1 and NA respectively. The relevance of using fresh versus dried Nopalea cochenillifera pad extracts against in vitro microbial growth is discussed.