Abstract Background: Mothers are crucial in the treatment and care of infants with hydrocephalus, as meeting the infant's demands requires daily effort. Hence, this study aimed to evaluate the effect of instructional guidelines on the challenges and adaptation strategies of mothers for caring for their infants with hydrocephalus. A pre-experimental design was used in this study. Setting: The study was conducted at Abo-Elrish-Elmonira Hospital affiliated with Cairo University Hospitals. Subject: A purposive sample that involved 120 mothers of hydrocephalus infants. Tools: Tool I: A structured questionnaire that included three parts (mothers' socio-demographic characteristics, mothers' knowledge, and practices) and tool II the challenges facing mothers in caring for their infants with hydrocephalus. Results: This study displayed a highly statistically significant difference between the pre and post – instructional guidelines among mothers' total knowledge, practice, and challenges regarding hydrocephalus (P value = 0.001). Conclusion: Instructional guidelines were important in addressing the challenges and adaptation strategies for mothers caring for their infants with hydrocephalus. Recommendation: Provide ongoing education and training that help mothers in caring for their infants with hydrocephalus.