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In this study, a total of 2674 fecal samples were collected from buffaloes (\n N<\/jats:italic>\n \u2009=\u2009875), cattle (\n N<\/jats:italic>\n \u2009=\u2009770), sheep (\n N<\/jats:italic>\n \u2009=\u2009520), and goats (\n N<\/jats:italic>\n \u2009=\u2009509) from different age groups across five governorates. All samples were examined using conventional microscopical methods. Using microscopy,\n Giardia<\/jats:italic>\n and\n Entamoeba<\/jats:italic>\n infections were detected in 626\/2674 (23.4%) and 457\/2674 (17.1%) of samples, respectively, and sheep showed the highest infection rates by both parasites (44.6% and 28.8% for\n Giardia<\/jats:italic>\n and\n Entamoeba,<\/jats:italic>\n respectively). Representative isolates were molecularly characterized, to confirm parasite identity prior to antigen preparation, targeting\n \u03b2-giardin<\/jats:italic>\n (\n bg<\/jats:italic>\n ) and small subunit ribosomal RNA (\n ssu rRNA<\/jats:italic>\n ) gene, confirmed the presence of\n G. duodenalis<\/jats:italic>\n (GenBank:\n OQ388331.1<\/jats:ext-link>\n ,\n OQ388332.1<\/jats:ext-link>\n ,\n OR029977.1<\/jats:ext-link>\n ,\n OR029976.1<\/jats:ext-link>\n ,\n OR029974.1<\/jats:ext-link>\n ,\n OR029975.1<\/jats:ext-link>\n and\n PP316110.1<\/jats:ext-link>\n ) and\n E. histolytica<\/jats:italic>\n (GenBank:\n OR016432.1<\/jats:ext-link>\n ). Antigens were prepared separately from these isolates, and the polyclonal antibodies (pAbs) were prepared in rabbits and purified using Protein-A-Sepharose affinity columns. Then, Anti\u2011\n Giardia<\/jats:italic>\n IgG pAbs and Anti-\n Entamoeba<\/jats:italic>\n IgG pAbs in-house sandwich ELISAs were tested separately on all fecal samples revealing infection rates of 45.2% (1208\/2674) for\n Giardia<\/jats:italic>\n sp. and 30.1% (805\/2674) for\n Entamoeba<\/jats:italic>\n sp. with high ability to detect\n Giardia<\/jats:italic>\n (\n \u03c7<\/jats:italic>\n \n 2<\/jats:italic>\n <\/jats:sup>\n \u2009=\u2009281.2,\n p<\/jats:italic>\n \u2009<\u2009\n 0.001<\/jats:italic>\n ) and\n Entamoeba<\/jats:italic>\n (\n \u03c7<\/jats:italic>\n \n 2<\/jats:italic>\n <\/jats:sup>\n \u2009=\u2009125.6,\n p<\/jats:italic>\n \u2009<\u2009\n 0.001<\/jats:italic>\n ) species compared to microscopy. In conclusion, the developed sandwich ELISA in our study represents a sensitive and reliable diagnostic tool for the detection of\n Giardia<\/jats:italic>\n and\n Entamoeba<\/jats:italic>\n infections in farm animals. Its application in routine surveillance along with complementary molecular confirmation could help to improve epidemiological accuracy and assess zoonotic risks.\n <\/jats:p>","DOI":"10.1186\/s12917-026-05801-4","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T11:34:57Z","timestamp":1786707297000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Giardia duodenalis and Entamoeba spp.: Coprological and antibody\u2013based approaches across farm animals in Egypt"],"prefix":"10.1186","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9718-5130","authenticated-orcid":false,"given":"Nagwa I.","family":"Toaleb","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0723-7276","authenticated-orcid":false,"given":"Kadria N.","family":"Abdel Megeed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2581-6503","authenticated-orcid":false,"given":"Dina","family":"Aboelsoued","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,14]]},"reference":[{"key":"5801_CR1","unstructured":"Abd El Hafez SM, Anwar AM, Ibrahim AM, Mahmoud MB, Hassan HM. 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