This study was performed to investigate the effects of lignocellulose supplementation (
LS
) on performance parameters, egg quality, aerobic bacterial load of eggshell, serum biochemical parameters, and jejunal histomorphological traits of laying hens between 18 and 38 wk of age. A total of 640 pullets at 16 wk of age were allotted to 4 treatment groups as 0 kg (control,
CONT
), 0.5 kg, 1 kg, and 2 kg LS per ton of feed. Body weight (
BW
), daily feed intake, egg production (
EP
), egg weight (
EW
), and efficiency of feed utilization (
EF
) were determined as the mean of each 3-wk period between 18 and 38 wk of age. Laying hens in the 1 kg LS group had a higher BW mean (1632.1 g,
P
< 0.001). The highest mean value of EP and EW were observed in 1 kg LS group (81.8% and 57.3 g, respectively), whereas the lowest values were found in the 2 kg LS group (78.6% and 54.4 g, respectively,
P
< 0.001). The mean of EF was the lowest in the 1 kg LS group (2.72,
P
< 0.001). There was a decline in eggshell breaking strength and eggshell thickness in the 2 kg LS, when compared with the 0.5 and 1 kg LS groups (
P
< 0.001). The total aerobic bacterial load of the eggshell was the lowest in the 1 kg LS group (4.7 log
10
cfu/mL). The level of aspartate amino transferase and alanine amino transferase showed an increment in both the CONT and 2 kg LS groups (
P
< 0.001). The high level of LS (2 kg per ton of feed) caused a decline in the levels of IgY, IgA and IgM, when compared to the 0.5 and 1 kg LS groups (
P
< 0.001). Laying hens in 0.5 and 1 kg LS groups had longer villus height (1335.9 μm) in the jejunum than the others (
P
< 0.001). These findings showed that the 1 kg LS per ton of feed improved EP and EW, eggshell quality, immunoglobulin levels and intestinal morphology, and decreased the total aerobic bacterial load.