Background: As addressing the demands of an infant with hydrocephalus necessitates daily effort, mothers play a critical role in the treatment and care of these infants. Hence, this study aimed to explore the effect of educational guidelines on knowledge-reported practices, and obstacles facing mothers having infants with hydrocephalus. A quasi-experimental design was used to achieve the aim of this study. Setting: The current study was applied in the pediatric neurosurgery department at Sohag University Hospital. Subject: A purposive sample that involved 100 mothers having infants with hydrocephalus. Tools: Tool I: A structured interview questionnaire that included three parts (part 1: mothers' characteristics, part 2: mothers' knowledge about hydrocephalus, and part 3: mothers' reported practices regarding hydrocephalus) and tool II: Obstacles facing mothers having infants with hydrocephalus in caring. Results: This study displayed a highly statistically significant difference between the pre and post-instructional guidelines among mothers' total knowledge, reported practices, and obstacles facing mothers regarding their infants with hydrocephalus (P value = 0.001). Conclusion: educational guidelines have positive effects in improving knowledge, reported practices, and obstacles facing mothers having infants with hydrocephalus. Recommendation: Provide ongoing education and training that help mothers having infants with hydrocephalus to get better their knowledge, and practices and overcome the obstacles faced during caring for their infants who suffer hydrocephalus.