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Lifetime childhood asthma prevalence (LCAP) percentages in Puerto Rico Health Regions (HR) are substantially higher in northeastern vs. southwestern HR. Higher average relative humidity in the northeast might promote mold and mite exposures and possibly asthma prevalence. To test this hypothesis, mold contamination, Environmental Relative Moldiness Index (ERMI) values were measured in floor dust (n=26) and dust mite allergen concentrations in bed dust (n=14). For this analysis, the eight HR were divided into those with LCAP >30% (n=3) and <30% (n=5). The average ERMI value was significantly greater (Wilcoxon Rank Sum, p<0.001) in high than in low LCAP HR (14.5 vs. 9.3). The dust mite antigens Der p 1, Der f 1, and Blo t 5 were detected in 90% of bed samples, but the concentrations were not significantly different in high vs. low LCAP HR. Mold exposures might partially explain the differences in LCAP HR in Puerto Rico.