IntroductionHaemonchus spp. are considered the most important strongylid nematodes with a worldwide distribution. The parasite’s blood-sucking nature can lead to severe anemia in infected animals. Despite its widespread impact, there is a dearth of comprehensive data on morphological and molecular identification methods for Haemonchus spp. in sheep from Upper Egypt. To address this gap, our current study aimed to assess the prevalence of Haemonchus spp. in 400 sheep fecal samples.MethodsWe employed microscopic examination and molecular techniques, using polymerase chain reaction (PCR) targeting the 18S gene for precise identification. Additionally, the potential risk factors associated with the infection by the parasite in sheep were explored.ResultsThe study pointed out that 33.00% (132 of 400) of the examined sheep were infected with Haemonchus spp. Sheep age and seasonal variability were found to be significant factors (p < 0.05) associated with the infection. Notably, sheep under 2 years old exhibited a higher risk, with an infection rate of 43.75% (84 out of 192), than their older counterparts. Furthermore, all reported infections were exclusively observed during the cold season, constituting 58.93% (132 out of 224) of cases. By contrast, no statistically significant association (p > 0.05) was found between the sex of the examined sheep and the occurrence of haemonchosis. Employing molecular methods, we isolated and identified the parasite through PCR analysis of cultured larvae, which were then subsequently confirmed as Haemonchus contortus via phylogenetic analysis.DiscussionThe study concluded that there was a relatively high occurrence of H. contortus among sheep from Upper Egypt. We recommend the implementation of stringent and effective control measures to combat the infection and safeguard livestock health.