In response to concerns about the stock status of bigeye tuna (BET) in the Eastern Pacific Ocean, the Inter-American Tropical Tuna Commission adopted additional management measures for BET in 2021, including an individual-vessel catch threshold system for the purse-seine fishery. Development of an enhanced port-sampling program for estimation of trip-level BET catch was also mandated to support member countries and their purse-seine vessels in their conservation efforts. To this end, a pilot study was conducted in the ports of Manta and Posorja, Ecuador, in 2022. In these ports, well unloading often takes place with small containers, which are used to transport fish from inside the well to the vessel’s wet deck. Using a high-frequency systematic sampling protocol, 63 wells of 37 trips with catch exclusively from floating-object (OBJ) sets were sampled from the start to the end of unloading, at a fixed interval from a random starting point, sampling about 10 % of all the containers of fish unloaded from each well. Notable large-scale pattern in the proportion of BET per container was found for 38 of the 56 wells that had BET catch. The proportion of BET per container, which could be high at the beginning, middle or end of the well unloading, sometimes varied by 0.40 or more. The magnitude of this large-scale within-well pattern had a significant increasing relationship with the number of OBJ sets associated with the well catch, consistent with variability in species composition among OBJ sets being an important factor in determining variability in species composition during catch unloading. Simulation studies demonstrated that such large-scale within-well pattern has potential implications for OBJ-set well-level sampling design, where, as expected from sampling theory, a systematic sampling protocol for containers of fish generally resulted in lower mean squared error (MSE) on the proportion of BET in the well, compared to simple random sampling of containers of fish. The reduction in MSE was largely due to a reduction in variance. Simulations also showed that the within-well variance component of the trip-level estimator for the proportion of BET can be larger than the among-well variance component, unless the within-well sampling coverage is relatively high. To minimize the negative impact of within-well variability on the precision of the estimator, results indicate that for a sampling protocol with one systematic sample per well, the within-well coverage should be at least 2 % of containers of fish unloaded from the well, and preferably more than 3 %. Finally, simulation studies were conducted with observer well plan data from 2013 to 2022 to put the well-level results in a larger context. Results indicated that the level of within-well sampling coverage may affect the variance on fleet-level estimates of the proportion of BET in the catch, with potentially substantially higher variance when the within-well sampling coverage is low. Taken together, these results suggest that both the type of within-well sampling and the level of within-well sampling coverage have important implications for the variance on BET catch estimates, from the well level to the fleet level.