In the present study, a trail to evaluate of Newcastle Disease (ND) antibodies levels after different vaccination programs was conducted on layer chickens. A total of 200 one day-old layer chicks (White Lohmann) were divided into five groups A, B, C, D and E. Birds in groups A, B and C were vaccinated with live vaccine by intraocular, intranasal and drinking water methods , respectively. On the other hand, groups D and E were kept as unvaccinated control groups. Vaccination performed at days 5, 18 and 28 by different routes for mentioned groups. Hemagglutination inhibition (HI) and enzyme-linked immunosorbent assay (ELISA) methods were used for assessment of antibodies titer at days 15, 25, 37 and 45. Results of HI and ELISA tests indicated that, the intranasal and the intraocular method have highest antibodies titers compared with the drinking water method. In this study, maternally derived antibodies specific to Newcastle Disease Virus (NDV) (IgY) were extracted by dextran sulfate method from collected eggs of vaccinated laying chickens . Antibodies specific to NDV (IgY) were detected in the egg yolk using HI test . Data revealed that antibodies specific to NDV (IgY) were presented in high titers that confer protection during early weeks of life for hatching chicks. Data concluded that extraction of maternally derived specific antibodies from egg yolk will facilitate accurate monitoring of ND vaccination programmes.