Three different forms of bacteria were used in this
study being incorporated in the diet either as live spray,
heat-inactivated form or freeze-dried form. The live
bacteria, harvested from a culture was heat-killed in a
water bath at temperature 75 8C for 60 min with
continuous stirring. The non-viability was checked by
plating on MRS agar three times. The freeze-dried
form was prepared by keeping the bacterial suspension
for 60 h at 20 8C in a REL 206 freeze-drier (Kyowa
Vacuum Tech., Tokyo, Japan). The freeze-dried
powder and suspension form was enumerated for
bacterial number per gram and milliliter of the product,
respectively. The freeze-dried form was vacuum
packed before preserving at 20 8C until further use.