Markets importing citrus fruit including lemons, Citrus limon (L.) Burman f., from South Africa require that the fruit be free of fruit fly pests (Diptera: Tephritidae). Historically there has been no fruit fly infestation recorded on lemons destined for export from South Africa. In this study, we assessed the host status of commercial export grade Eureka lemons, Citrus limon (L.) Burmanf. cv. Eureka, for four fruit fly pest species of economic importance in South Africa: Ceratitis capitata (Wiedemann), Ceratitis rosa Karsch, Ceratitis quilicii De Meyer, Mwatawala & Virgilio, and Bactrocera dorsalis (Hendel). Trapping was conducted in 10 Eureka lemon orchards in two major citrus production regions over two citrus seasons between 2016 and 2017 to determine the level of fruit fly abundance in the sampled orchards. Lemons were collected at harvest over the two seasons in the same orchards where trapping was conducted. Fruit fly infestation of the sampled lemons was determined by dissection. Additionally, infestation of lemons was determined under forced exposure to mature mated females of C. capitata and B. dorsalis. Trapping data showed the presence of adults of all four fruit fly species in the sampled lemon orchards. No fruit fly infestation was detected in 43 222 Eureka lemons sampled at harvest. There was also no infestation of lemons under forced exposure conditions. The results of this study provide evidence with 99.99 % efficacy and a 99%confidence level that SouthAfrican commercial export grade Eureka lemon fruit is not a host for C. capitata, C. rosa, C. quilicii or B. dorsalis.