People who inject drugs (PWID) are at higher risk of acquiring HIV due to risky injection and sexual practices. We measured HIV prevalence and behaviors related to acquisition and transmission risk at two time points (2007 and 2012) in Zanzibar, Tanzania. We conducted two rounds of behavioral and biological surveillance among PWID using respondent-driven sampling, recruiting 499 and 408 PWID, respectively. Through faceto- face interviews, we collected information on demographics as well as sexual and injection practices. We obtained blood samples for biological testing. We analyzed data using RDSAT and exported weights into STATA for multivariate analysis. HIV prevalence among sampled PWID in Zanzibar was 16.0 % in 2007 and 11.3 % in 2012; 73.2 % had injected drugs for 7 years or more in 2007, while in the 2012 sample this proportion was 36.9 %. In 2007, 53.6 % reported having shared a needle in the past month, while in the 2012 sample, 29.1 % reported having done so. While 13.3 % of PWID in 2007 reported having been tested for HIV infection and received results in the past year, this proportion was 38.0 % in 2012. Duration of injection drug use for 5 years or more was associated with higher odds of HIV infection in both samples. HIV prevalence and indicators of risk and preventive behaviors among PWID in Zanzibar were generally more favorable in 2012 compared to 2007-a period marked by the scale-up of prevention programs focusing on PWID. While encouraging, causal interpretation needs to be cautious and consider possible sample differences in these two cross-sectional surveys. HIV prevalence and related risk behaviors persist at levels warranting sustained and enhanced efforts of primary prevention and harm reduction.