* purpose: to identify spore forming bacteria from non spore forming bacteria -it is a differential stain -not all bacteria produce spores -spores are means for survival not reproduction

- spores are produce in unfavorable conditionsuch as: 1- lack of nutrients 2-suden change of temp. 3- change of pH 4- dryness

-spores found inside the bacteria are called Endospores - Endospores have different location in bacteria: 1-terminal 2- subterminal

3-central

*Life cycle of spores:

*sporulation:is the trasfer or change of bacteria (vegetative cell ) to endospore in unfavorable condition * germination:the transfer or change of spore to bacteria in favorable condition

Spore stain procedure: 1- Put one drop of water in the middle of plain slide 2- Inculcate the bacteria 2- Inculcate the bacteria 3- Do the smearing 3- Do the smearing 4- Allow to air dry 4- Allow to air dry 5- Do heat fixation for 3x 5- Do heat fixation for 3x

1- malachite green (8-10min) 2- wash with water 3- safranin(1min) 4- wash with water 5- dry then examine under the microscope