At 28 days of age, 20 birds (5 birds/treatment)
were randomly selected and bled, then
slaughtered to sample the small intestine.
Small intestine samples were washed with a
solution (formalin 90% and distilled water
10%), then they were stored in a chiller at 4°C.
Small intestine samples were obtained for histology
measurement. Briefly, duodenum and
jejunum were cut into segments of 5 mm of
length, placed in cassettes and soaked in formalin
and alcohol of different concentrations
using Histokinette device 2000 (Leica
Microsystems GmbH, Wetzlar, Germany) for
the fixation and dehydration of intestinal tissues.
Then dehydrated tissues were covered
with paraffin at 75°C using a paraffin dispenser
while still in cassettes. After that, prepared
paraffin cassettes were left cooling at
room temperature, then stored at freezer temperature
until the time of cutting and slides
preparation. Paraffin cassettes were cut using
a microtome device at a thickness of 5 µm.
Multiple slides were prepared from each tissue
and all slides were stained using a PAS stain
(Sheehan and Hrapchak, 1980).