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Background We estimated the prevalence of colonization with extended-spectrum cephalosporin-resistant Enterobacterales (ESCrE) and carbapenem-resistant Enterobacterales (CRE) from a hospital and associated communities in western Guatemala. Methods Randomly selected infants, children, and adults (<1, 1–17, and ≥18 years, respectively) were enrolled from the hospital (n = 641) during the coronavirus disease 2019 (COVID-19) pandemic, March to September 2021. Community participants were enrolled using a 3-stage cluster design between November 2019 and March 2020 (phase 1, n = 381) and between July 2020 and May 2021 (phase 2, with COVID-19 pandemic restrictions, n = 538). Stool samples were streaked onto selective chromogenic agar, and a Vitek 2 instrument was used to verify ESCrE or CRE classification. Prevalence estimates were weighted to account for sampling design. Results The prevalence of colonization with ESCrE and CRE was higher among hospital patients compared to community participants (ESCrE: 67% vs 46%, P <.01; CRE: 37% vs 1%, P <.01). Hospital ESCrE colonization was higher for adults (72%) compared with children (65%) and infants (60%) (P <.05). Colonization was higher for adults (50%) than children (40%) in the community (P <.05). There was no difference in ESCrE colonization between phase 1 and 2 (45% and 47%, respectively, P >.05), although reported use of antibiotics among households declined (23% and 7%, respectively, P <.001). Conclusions While hospitals remain foci for ESCrE and CRE colonization, consistent with the need for infection control programs, community prevalence of ESCrE in this study was high, potentially adding to colonization pressure and transmission in healthcare settings. Better understanding of transmission dynamics and age-related factors is needed. [ABSTRACT FROM AUTHOR]