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Background: Chagas disease (CD) is a chronic parasitic disease caused by Trypanosoma cruzi and is endemic to continental Latin America. In Spain, the main transmission route is congenital. We aimed to assess adherence to regional recommendations of universal screening for CD during pregnancy in Latin American women in the province of Alicante from 2014 to 2018. Methodology/Principal findings: Retrospective quality study using two availa sources: 1) delivery records of Latin American women that gave birth in the 10 public hospitals of Alicante between January 2014 and December 2018; and 2) records of Chagas serologies carried out in those centers between May 2013 and December 2018. There were 3026 deliveries in Latin American women during the study period; 1178 (38.9%) underwent CD serology. Screening adherence ranged from 17.2% to 59.3% in the different health departments and was higher in Bolivian women (48.3%). Twenty-six deliveries (2.2%) had a positive screening; CD was confirmed in 23 (2%) deliveries of 21 women. Bolivians had the highest seroprevalence (21/112; 18.7%), followed by Colombians (1/333; 0.3%) and Ecuadorians (1/348; 0.3%). Of 21 CD-positive women (19 Bolivians, 1 Colombian, 1 Ecuadorian), infection was already known in 12 (57.1%), and 9 (42.9%) had already been treated. Only 1 of the 12 untreated women (8.3%) was treated postpartum. Follow-up started in 20 of the 23 (87.0%) neonates but was completed only in 11 (47.8%); no cases of congenital transmission were detected. Among the 1848 unscreened deliveries, we estimate 43 undiagnosed cases of CD and 1 to 2 undetected cases of congenital transmission. Conclusions/Significance: Adherence to recommendations of systematic screening for CD in Latin American pregnant women in Alicante can be improved. Strategies to strengthen treatment of postpartum women and monitoring of exposed newborns are needed. Currently, there may be undetected cases of congenital transmission in our province. Author summary: Chagas disease (CD) is a neglected tropical disease endemic to Latin America. In absence of the triatomine vector in Spain, congenital (mother-to-infant) transmission is the main infection route. The Valencian Community has recommended universal screening for CD in pregnant Latin American women since 2007. In our study we analyzed adherence to that recommendation in Alicante province from 2014 to 2018, finding that it is quite low (38.9% overall, 48.3% in Bolivians) and heterogeneous between health departments. Among unscreened pregnant women during the study period, we estimate that there could be 43 undiagnosed cases of CD and 1 to 2 undetected infections in infants. We also observed very low adherence to treatment after delivery in CD-diagnosed, untreated women (8.3%), and a low rate of completed follow-up in newborns at risk of vertical infection (47.8%). We need to improve the program in order to achieve universal CD screening in Latin American (and especially Bolivian) pregnant women, to enhance CD treatment in postpartum women, and to improve monitoring in exposed newborns through a well-established notification and follow-up circuit. [ABSTRACT FROM AUTHOR]