Fig 1.
Laying curve (A) and number of double-yolk eggs (B) in broiler hens from 23rd to 40th week. Eggs were collected and weighed daily for all hens fed with different concentrations of GSE dietary supplementation or with a control diet. A: animals fed with control diet without GSE supplementation (n = 92), B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old (n = 80), and D: supplementation with GSE at 1% of the total diet composition starting at hatch until 40 week-old (n = 72). Results are presented as lsmeans ± S.E.M. P values and different individual letters indicate a significant effect of the diet between A, B and C groups or between A and D groups. P value was considered as significant if P < 0.05.
Table 1.
Number and quality of laying eggs of broiler hens fed with a GSE diet supplemented.
Group A: animals fed with control diet without GSE supplementation, B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old and D: supplementation at 1% of the total diet composition starting at hatch until 40 week-old.
Fig 2.
Weight of preovulatory follicles F1 (A), F2 (B), F3 (C) and F4 (D) of broiler hens fed with different concentrations of GSE supplementation.
Follicular hierarchy of 10 animals selected randomly from each group (A to D) was analysed at the 40th week. Each preovulatory follicle from each animal was weighed. A: animals fed with control diet without GSE supplementation, B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old, and D: supplementation with GSE at 1% of the total diet composition starting at hatch until 40 week-old. Results are presented as lsmeans ± S.E.M. P values and different individual letters indicate a significant effect of the diet between A, B and C groups (experiment 1) or between A and D (experiment 2) groups. P value was considered as significant if P < 0.05.
Table 2.
Percentage of unfertilised eggs, early (EEM) and late (LEM) embryonic mortality and fertility after artificial insemination in broiler hens fed with GSE supplementation.
Fig 3.
Levels of progesterone (A), testosterone (B), androstenedione (C) and oestradiol (D) in egg yolk and P450 aromatase (E) and StAR (F) mRNA expression in granulosa cell from broiler hens fed with different concentrations of GSE.
Steroids were extracted from 30 egg yolks for each group, at 33 weeks-old. mRNA was extracted from granulosa cells of 10 preovulatory follicles for each group. A: animals fed with control diet without GSE supplementation, B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old (experiment 1) and D: supplementation with GSE at 1% of the total diet composition starting at hatching until 40 week-old (experiment 2). Results are presented as lsmeans ± S.E.M. P values and different individual letters indicate a significant effect of the diet between A, B and C groups (experiment 1) or between A and D groups (experiment 2). P value was considered as significant if P < 0.05. **P < 0.005 and **** P < 0.0001.
Table 3.
Progesterone, testosterone, androstenedione and oestradiol concentrations in egg yolk at 33 weeks-old.
Fig 4.
Effect of dietary GSE supplementation on in vitro steroidogenesis at basal state (A) and in response to IGF1 and LH alone or in combination in hen primary granulosa cells.
Granulosa cells from preovulatory follicles 1 (F1) from different group of animals (A to D) were seeded for 24 h and after overnight serum starvation, granulosa cells were incubated with IGF1 (10-8M), LH (10-8M) or both for 48 h. The culture medium was then collected. Basal progesterone level (A) and the response to IGF1 or/and LH (B) were assessed. A: animals fed with control diet without GSE supplementation, B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old, and D: supplementation with GSE at 1% of the total diet composition starting at hatch until 40 week-old. Results are presented as lsmeans ± S.E.M. P values and different individual letters indicate a significant effect of the diet between A, B and C groups (experiment 1) or between A and D groups (experiment 2). P value was considered as significant if P < 0.05. **P < 0.005 and **** P < 0.0001.
Fig 5.
Level of Reactive Oxygen Species (ROS, H202) in egg yolk of broiler hens fed with different concentrations of GSE dietary supplementation.
ROS level (H202) was assessed from 1 g of each egg yolk using Ros-Glo assay. Ten egg yolks were analysed for each group of animals. A: animals fed with control diet without GSE supplementation, B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old (experiment 1), and D: supplementation at 1% of the total diet composition starting at the hatching until 40 week-old (experiment 2). Results are presented as lsmeans ± S.E.M. P values and different individual letters indicate a significant effect of the diet between A, B and C groups (experiment 1) or between A and D groups (experiment 2). P value was considered as significant if P < 0.05. **** P < 0.0001.
Fig 6.
Level of plasma RARRES2 (A), ADIPOQ (B) and NAMPT(C) concentrations of broiler hens fed with different concentrations of GSE dietary supplementation.
At 33rd week, plasma adipokines concentration was assessed using specific ELISA assay in 11 animals from each group of animals. A: animals fed with control diet without GSE supplementation, B and C: animals supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, starting at 4 week-old until 40 week-old, and D: supplementation at 1% of the total diet composition starting at hatch until 40 week-old. Results are presented as lsmeans ± S.E.M. P values and different individual letters indicate a significant effect of the diet between A, B and C groups (experiment 1) or between A and D groups (experiment 2). P value was considered as significant if P < 0.05. **** P < 0.0001.
Table 4.
Pearson correlation coefficient (r) calculated between plasma adipokines (RARRES2, ADIPOQ, NAMPT) concentration and steroids hormones (progesterone, androstenedione, testosterone, oestradiol) and ROS concentration in egg yolk, for the groups A, B and C (experiment 1) and then, A and D (experiment 2).
Table 5.
mRNA expression of adipokines (RARRES2, ADIPOQ, NAMPT) and their receptors (CMKLR1, CCRL2, GPR1, ADIPOR1, ADIPOR2) in granulosa and theca cells from preovulatory follicle 1 (F1).