الفهرس 
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Abstract
Objectives:
Natural feed additives are gaining interest as safe alternative growth promoters and potential immune
stimulants instead of antibiotics. This study aimed to investigate the immunomodulatory influence of
Nannochloropsis oculata (N. oculata) and olive leaf extract (OLE) in vitro and in vivo in pregnant ewes and
lambs.
Methods:
In vitro lymphocyte culturing was carried out using Peripheral blood mononuclear cells (PBMCs).
PBMCs wells were sectioned into 7 groups; group I (C): considered as control, group II (N 250): treated with N. oculate extract at 250 µg/mL, group III (N 500): treated with N. oculate extract at 500 µg/mL,
group IV (N 1000): treated with N. oculate extract at 1000 µg/mL, group V (OLE 250): treated with OLE
at 250 µg/mL, group VI (OLE 500): treated with OLE at 500 µg/mL, and group VII (OLE 1000): treated
with OLE at 1000 µg/mL in 1% DMSO.
Meanwhile, in vivo 24 healthy pregnant ewes (2-3 years, 45-55 Kg) and 24 healthy male Barki lambs
(3-6 months, 19-24 Kg) were divided into 4 groups; group 1: fed with basal diet (C), group 2: animals fed
basal diet supplemented with N. oculata at 3% (N 3%), group 3: animals fed basal diet supplemented with
olive leaf extract at 2% (OLE 2%), and group 4: animals fed basal diet supplemented with olive leaf extract
at 4% (OLE 4%). Experiments was extended for one month.
The body weight of pregnant ewes and lambs were measured at the start and the end of the
experiment, blood samples were collected at the end of the experiment, and feed intake was measured
weekly. In vitro and in vivo evaluations of lymphocyte transformation, INF-γ, TNF-α, IL-4, IL-10, and IL-12
were performed, as well as in vivo serum MDA and SOD.
Results:
Lymphocyte Transformation was greatly enhanced in vitro when incubated with N. oculata extract
and OLE at concentration 1000 µg/mL, while lymphocyte transformation was significantly reduced when
incubated with 250 µg/mL of OLE. All concentrations of N. oculata extract and OLE significantly (P <0.05)
elevated the levels of cytokines such as INF-γ, TNF-α, IL-4, IL-10, and IL-12 in vitro study in presence of
Pokeweed mitogen (P.K). In vivo study, both N. oculata and OLE significantly (P <0.05) improved the body
weight gain in growing lambs and lowered the feed conversion ratio in both pregnant ewes and growing
lambs than the control group. There was no significant difference between groups at the hematological
parameters in lambs. N 3% and OLE 2% significantly (P <0.05) increased the lymphocyte transformation in
vivo in presence of P.K compared to the control group in pregnant ewes and growing lambs. TNF-α, IL-4, IL10, and IL-12 showed a significant difference in vivo among treated and untreated groups in both pregnant
ewes and growing lambs. All supplemented groups significantly (P <0.05) reduced serum MDA levels and
enhanced the serum SOD levels in pregnant ewes and growing lambs compared to the control group.