Aerial surveys are often used to quantify sizes of waterbird colonies; however, these surveys would benefit from a better understanding of associated biases. We compared estimates of breeding pairs of waterbirds, in colonies across southern Louisiana, USA, made from the ground, fixed-wing aircraft, and a helicopter. We used a marked-subsample method for ground-counting colonies to obtain estimates of error and visibility bias. We made comparisons over 2 sampling periods: 1) surveys conducted on the same colonies using all 3 methods during 3–11 May 2005 and 2) an expanded fixed-wing and ground-survey comparison conducted over 4 periods (May and Jun, 2004–2005). Estimates from fixed-wing aircraft were approximately 65% higher than those from ground counts for overall estimated number of breeding pairs and for both dark and white-plumaged species. The coefficient of determination between estimates based on ground and fixed-wing aircraft was ≤0.40 for most species, and based on the assumption that estimates from the ground were closer to the true count, fixed-wing aerial surveys appeared to overestimate numbers of nesting birds of some species; this bias often increased with the size of the colony. Unlike estimates from fixed-wing aircraft, numbers of nesting pairs made from ground and helicopter surveys were very similar for all species we observed. Ground counts by one observer resulted in underestimated number of breeding pairs by 20% on average. The marked-subsample method provided an estimate of the number of missed nests as well as an estimate of precision. These estimates represent a major advantage of marked-subsample ground counts over aerial methods; however, ground counts are difficult in large or remote colonies. Helicopter surveys and ground counts provide less biased, more precise estimates of breeding pairs than do surveys made from fixed-wing aircraft. We recommend managers employ ground counts using double observers for surveying waterbird colonies when feasible. Fixed-wing aerial surveys may be suitable to determine colony activity and composition of common waterbird species. The most appropriate combination of survey approaches will be based on the need for precise and unbiased estimates, balanced with financial and logistical constraints.