Waterhyacinth is a free-floating aquatic weed that is considered a nuisance worldwide. Excessive growth of waterhyacinth limits recreational use of water bodies as well as interferes with many ecological processes. Accurate estimates of biomass are useful to assess the effectiveness of control methods to manage this aquatic weed. While large water bodies require significant labor inputs with respect to ground-truth surveys, available technology like remote sensing could be capable of providing temporal and spatial information from a target area at a much reduced cost. Studies were conducted at Lakes Columbus and Aberdeen (Mississippi) during the growing seasons of 2005 and 2006 over established populations of waterhyacinth. The objective was to estimate biomass based on nondestructive methods using the normalized difference vegetation index (NDVI) derived from Landsat 5 TM simulated data. Biomass was collected monthly using a 0.10m2 quadrat at 25 randomly-located locations at each site. Morphometric plant parameters were also collected to enhance the use of NDVI for biomass estimation. Reflectance measurements using a hyperspectral sensor were taken every month at each site during biomass collection. These spectral signatures were then transformed into a Landsat 5 TM simulated data set using MatLab® software. A positive linear relationship (r2 = 0.28) was found between measured biomass of waterhyacinth and NDVI values from the simulated dataset. While this relationship appears weak, the addition of morphological parameters such as leaf area index (LAI) and leaf length enhanced the relationship yielding an r2 = 0.66. Empirically, NDVI saturates at high LAI, which may limit its use to estimate the biomass in very dense vegetation. Further studies using NDVI calculated from narrower spectral bands than those contained in Landsat 5 TM are recommended.