<p>Tree-related microhabitats (TreMs), particularly treeholes, are important but understudied components of tropical landscapes that support diverse invertebrate communities. Baseline information on treehole-associated invertebrates is particularly scarce in human-dominated environments worldwide, and this knowledge gap is especially evident in Central America. This study provides the first baseline assessment of invertebrate abundance, diversity, and richness in treehole microhabitats across rural and urban areas of northern Belize, and evaluates preliminary relationships between invertebrate abundance and selected tree and treehole characteristics. Detritus samples were collected from 30 naturally occurring treeholes (15 rural and 15 urban) between 2023 and 2024. No statistically significant differences were detected in invertebrate abundance or composition between rural and urban treeholes (<i>P</i> &gt; 0.05). Rural treeholes supported higher species richness, whereas urban treeholes exhibited greater diversity and evenness. From the Test of Model Effects, treehole height was the only variable found to be individually significant. The other variables, treehole height above ground, treehole width, and treehole depth, showed weak and non-significant relationships with invertebrate abundance. According to the Pearson analysis of linear association, the invertebrate abundance per treehole did not exhibit any statistically significant linear relationship with the treehole characteristic predictor variables. The Directional Eta analyses showed that when treating invertebrate treehole abundance as the dependent variable, tree types (η = 1.000) showed the strongest association. The findings provide a baseline for future, larger-scale investigations examining the effects of anthropogenic activities on treehole invertebrate communities in human-dominated habitats.</p>