A prioritisation study was conducted to address the lack of adequate information about potential pests likely to be introduced in Zambia and become invasive. The study was conducted by subject matter experts from relevant institutions in and outside Zambia. Although this study focused on major pest categories, this paper only addresses bacteria and Protista. A list of 306 bacterial and 10 Protista species adjudged to affect plants was generated using CABI’s Horizon Scanning Tool. The 316 (total) pest species were refined to focus on pests that affect value chains important to Zambia’s economy. This resulted in a final list of 133 bacteria and eight Protista. Four additional bacteria species considered of phytosanitary interest were added and all 137 bacteria and eight Protista species were subjected to a rapid risk assessment using agreed guidelines. Vectors reported to transmit any of the pathogenic organisms were also subjected to a risk assessment. A proportion of 53% (n = 77 of 145) comprising 73 bacteria and four Protista species were reported as present in Africa. Of these, 42 (57%, n = 73) bacterial species and two (n=4) Protista species were reported in neighbouring countries. Considering a cut-off of 54, the highest scoring pests were 40 bacteria (highest score of 140) and three Protista (highest score of 125). Three actions were suggested for high-scoring pests, a detection surveillance, a pest-initiated pest risk analysis (PRA) or a detection surveillance followed by pest-initiated PRA. A “no action” was suggested where the risk was very low although, for some pathogenic organisms, a “no action” was followed by periodic monitoring. This information will contribute towards proactive prevention and management of biological invasions.