Seroprevalence and molecular characterization of Toxoplasma gondii infecting ruminants in the North-West of Egypt

•This study aimed to evaluate the molecular and seroprevalence of T. gondii in ruminants in Egypt•ELISA results were confirmed by a nested PCR targeting B1 gene and single PCR targeting P30 gene•Age, sex, species, and the place of the collection were considered risk factors for toxoplasmosis•PCR assays could ensure the infection in the blood successfully as good as ELISA•Phylogenetic analysis revealed diversity in submitted strains of T. gondii Toxoplasma gondii is a coccidian parasite known for its heavy toll on people and livestock. It can cause abortion and a variety of congenital diseases. The current study aimed to examine some seroprevalence and molecular attributes of T. gondii obtained from ruminants in the North-West of Egypt. Specimens were random selected from five different locations in Alexandria and Matrouh governorates. A total of 483 blood samples, collected from 96 mixed flocks, were screened for anti-T. gondii IgG antibodies using enzyme-linked immunosorbent assay (ELISA). The seropositive results were then confirmed using polymerase chain reaction (PCR) primers for the B1 and P30 genes. Specific PCR products were selected for sequencing and alignment against the GenBank, where phylogeny has been examined using the maximum likelihood, neighbor-joining, and maximum parsimony in MEGA6. ELISA confirmed the presence of T. gondii in 188 of the investigated samples (38.92%), indicating a higher prevalence in camels (64.51%) and sheep (43.75%) as compared to goats (27.93 %) and cattle (13.46%). PCR confirmed the presence of T. gondii-specific sequences in 159 seropositive specimens, with homology between 98.3 and 100%. The genetic distances between the investigated variants ranged from 0.1 to 0.9, and 7 single nucleotide polymorphisms (SNPs), were identified in the examined T. gondii specimens. The camel T. gondii parasite, isolated from Matrouh, showed a 100% homology with the most dangerous reference strains of T. gondii-RH in the GenBank. Our results showed that B1 and P30-specific PCR could detect T. gondii in blood samples more accurately than ELISA. In addition, the statistical analysis of our data indicated that species, age, sex, and animal location were all risk factors for toxoplasmosis. These findings are likely to boost disease control and help contain the spread of T. gondii infections. [Display omitted]