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Two similar to 40-cm-long sediment cores, PI-SC-1-10m and PI-SC-2-40m, were recovered at 10 and 40 m water depth, respectively, from Lago Peten Itza, in the Department of Peten, northern Guatemala. The cores span the last similar to 525 years of sediment accumulation in the basin. This study explores lake level and trophic state changes that Lago Peten Itza has experienced since European contact in the early 1500s. We inferred past environmental variability using changes in sediment geochemistry and fluctuations in relative species abundances of ostracode and chironomid fossil assemblages. Changes in concentrations of organic matter (OM), carbonate, total carbon (TC), total nitrogen (TN), C/N ratios, bromine (Br), and faunal relative abundances were used to infer changes in the trophic status of the lake. Cultural eutrophication began in the 1930s, and anthropogenic impact increased significantly after similar to 1970. Higher linear sedimentation rates, up to 9.1 mm yr(-1), began recently. They are attributed to increases in riparian settlement, deforestation and associated land use changes, which led to accelerated soil erosion. An ostracode-based transfer function was applied to assemblages in core PI-SC-1-10m, which enabled us to identify periods when lake level fluctuated. Such historical fluctuations in lake levels were driven primarily by changes in rainfall. Past lake levels can be summarized as follows: (1) fluctuating, high lake levels from similar to 1550s to the 1730s and from the early 1940s to 2005, and (2) stable, lower lake levels from about 1750 to the early 1900s. Higher relative abundance of the ostracode Physocypria globula and higher rubidium (Rb) concentrations indicate higher lake levels than today. Chironomids also show sharp fluctuations along the cores that could be related to water level changes. The presence of chironomid assemblages Chironomus, Procladius, and Einfeldia from 1960-2000 AD shows high productivity levels. The Lago Peten Itza sediment record indicates a generally arid Little Ice Age (LIA), with exceptions around 1580 and 1650 when high lake levels similar to those of the 20(th) century, i.e. similar to 5 m higher than today, indicate more humid conditions.