In vitro characterization and in ovo embryotoxicity assessment of a triazol-5-one derivative in broiler embryos

This study is among the pioneering investigations evaluating the embryonic effects of triazol-5-one derivatives administered in ovo to broiler embryos. The synthesized compound, 1-(morpholin-4-yl-methyl)-3-(p-chlorobenzyl)-4-(4-hydroxybenzylideneamino)-4,5-dihydro-1H-1,2,4-triazol-5-one, was examined for its in vitro biological properties and in ovo embryonic safety. The compound is hypothesized to influence embryonic viability in a manner consistent with its structural and biological characteristics. Structural confirmation was achieved through FT-IR, ¹H-NMR, and ¹³C-NMR analyses. In vitro assays showed weak reducing and radical-scavenging capacity but high, stable metal-chelating activity, alongside strong antimicrobial effects. For the in ovo trial, 120 fertilized Ross-308 eggs were assigned to three groups (Control, Dimethyl sulfoxide (DMSO), Experimental; n = 40). On day 17, 0.5 mL of solution was injected into the amniotic cavity, and embryos were monitored until 510 h. Hatchability was 87.5% in the Control group, 80% in the DMSO group, and 0% in the Experimental group. No live chicks hatched in the experimental group, indicating complete embryonic lethality. All embryo losses occurred after injection during incubation, and Levene's test followed by one-way ANOVA (p = 0.385) confirmed no initial weight differences, indicating a compound-specific effect. The compound's embryotoxicity may result from strong metal chelation, low antioxidant capacity, oxidative stress, mitochondrial dysfunction, and membrane disruption. These findings provide a basis for defining safe dose ranges, optimizing injection parameters, and assessing the in ovo applicability of related triazole compounds.