*4.7. Data Management and Statistical Procedures*

Data were double entered in Epi Info 3.5.1 (CDC, Atlanta, 2008, Atlanta, GA, USA) to minimize entry errors, followed by data comparison and inconsistencies resolution. Data were analyzed using IBM SPSS software (Statistical Package for the Social Science, Armok, NY: IBM Corp, 2011, version 26.0, Chicago, IL, USA). Categorical variables were summarized as frequencies and continuous variables were summarized as medians and Inter-quartile Range (IQR). Contingency tables were built between dependent and independent variables. Crude and adjusted odds ratio were estimated through simple and multiple logistic regression models. Independent variables with *p*-values ≤ 0.2 in the simple logistic regression were included in the multiple logistic regression model in order to obtain adjusted odds ratio. Goodness-of-fit was assesses using the Hosmer and Lemeshow test for the multiple logistic regression model. A p-value less than 0.05 was considered evidence of statistical significance and all analyses were performed considering a 95% confidence interval (95% CI).

#### **5. Conclusions**

This study showed a high frequency of *Cryptosporidium* spp. infection detected by PCR-RFLP among children admitted to two public hospitals in Maputo City due to acute diarrhea. Child age (25 to 60 months) and caregiver literacy status were predictors for infection. Our findings demonstrated that the infection is mostly due to *C. hominis*. This suggests mainly anthroponomic transmission, with great public health implications. The present study demonstrated the need for improved public health recommendations by including *Cryptosporidium* spp. in routine testing among children with diarrhea in Mozambique. Moreover, the study presented the need for an establishment of more accurate molecular characterization platform of the parasite in a national context, in order to understand which species occur in the country, routes of infection (if anthroponotic and/or zoonotic) and/or regional patterns, informing public health programs.

**Author Contributions:** Conceptualization, N.d.D. and I.C.-M.; methodology, I.C.-M. and H.C.; validation, I.C.-M., H.C., M.F., L.A.L.-O. and A.M.D.-C.; formal analysis, I.C.-M., H.C. and A.F.L.B.; Investigation, H.C.; resources, N.d.D., M.F., L.A.L.-O. and A.M.D.-C.; data curation, H.C.; writing original draft preparation, H.C.; writing—review and editing, H.C., I.C.-M. and A.F.L.B.; visualization, M.C., M.B., E.L.G., E.A., J.S., J.S.L., D.M.B., L.V.M.-C. and A.C.; supervision, N.d.D., A.M.D.-C. and I.C.-M.; project administration, J.C.; funding acquisition, N.d.D. All authors have read and agreed to the published version of the manuscript.

**Funding:** The current analysis was performed as part of the requisites for a master's degree and was supported in part by FIOCRUZ/IOC/RJ-Brazil and Instituto Nacional de Saúde—Moçambique post-graduation program. M.F. received funding from Universal/CNPq at and FAPERJ (Grant E-26/202.078/2020) and fellowship from CAPES (Brasil Sem Miséria/Brazilian governmental program), CNPq (PDJ), INOVA Program (FIOCRUZ), and FAPERJ. The National Surveillance for Acute Diarrhea in Children project was supported by a Senior Fellowship awarded to N.D. by the European Foundation Initiative for African Research into Neglected Tropical Diseases (EFINTD, Grant number: 89539) and World Health Organization (WHO).

**Institutional Review Board Statement:** The protocol was approved by the Mozambique National Bioethics Committee for Health (IRB00002657, reference Nr. 348/CNBS/13).

**Informed Consent Statement:** Informed consent was obtained from all subjects involved in the study.

**Data Availability Statement:** The data presented in this study are available on request from the corresponding author. The data are not publicly available due to the ethical reasons.

**Acknowledgments:** We would like to thank the caregivers who consented to their children being enrolled in the surveillance. We wish to thank the Sequencing Service DNA Sequencing Platform– Fundação Oswaldo Cruz for their contribution in the study procedures, Direcção Clínica de Pediatria do Hospital Geral de Mavalane and Hospital Geral José Macamo, and Carlos Guilamba and Celina Nhamuave for their amazing work on data entry. Finally, we would like to thank all the focal points in the hospital who helped conduct this study and the members of the surveillance team. We would also like to thank Kristen Heitzinger and Neha Kamat who reviewed the language in the manuscript.

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **References**

