NaturalGreen

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<b><span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;">Endospore</span></b><span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;"> is domanet strucure
which have no or less metabolic activities. Bactiers form endospre when
environemntal conditions are  unfavorable
to them such as  high oxygen
concentraion, temperature, heat and desssication. Endospore cannot be killed by
just boilling at 100◦C but at 121◦C. <b><i>Bacillus, Clostridium </i></b>and<b><i>
Sporalactobacillus</i></b><i> </i>are
examples for endospore froming bacteria<i>. <o:p></o:p></i></span></div>
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<i><span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;">                             </span></i><span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;">The outermost layer
of the endopsore is proteinaceous <b>coat</b>.
Bneeth coat, there is a a very thick layer of peptidoglycan which is called as the
<b>cortex</b>. Under cortex, <b>grem cell wall</b> reside. After endospire
germination, grem cell wall become the cell wall fo bacterium. Bneath gren cell
wall, <b>inner membrane</b> is founsd. It
act as adverse premeability barrier. The <b>core
</b>exsits in the center of the endopore. The core is the dehydreted DNA of the
bacterium cell.</span></div>
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<b><span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;">Endospore  staining</span></b><span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;"> is a differentiate staining
technique which is used to differentiate bacteria into two groups such as
endospore forming baetria and non endospore forming bateria. Endospore is
stained in green while vegetative cell is stained in pink. <o:p></o:p></span></div>
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<span style="font-size: 14.0pt; line-height: 115%; mso-no-proof: yes;"><b><span style="font-size: 14.0pt; line-height: 115%;">Position of endospore</span></b><span style="font-size: 14pt; line-height: 115%;">: The position of endospore inside
the vegetative cell varies from one bacterium species to another as follows.</span> <o:p></o:p></span></div>
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<b><span style="font-size: 14.0pt; line-height: 115%;">Procedure</span></b><span style="font-size: 14.0pt; line-height: 115%;">- A loopful from a broth or agar
plate is taken into a glass slide. Then the smear is prepared by heat fixing at
bunson flame. As the proteins of the cell wall are denatured by heat, it
supports the bacterium cells to adhere to the slide. The smear is stained by
flooding with Malachite green. It is allowed Malachite green to act for 5
minutes.  The underside of the slide is
heated by using alcohol deep cotton wool. 
It is allowed the slide to cool. Stained smear is decolorized by using
water and air dry. Safranin is added as the counterstain. It is allowed
Safranin to react for 30seconds. Excess Safranin is washed away by using water
and air dry. Endospore and it vegetative cell are observed under oil immersion
objectives.<o:p></o:p></span></div>
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<b><u><span style="font-size: 14.0pt; line-height: 115%;">Observations:<o:p></o:p></span></u></b></div>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjDemc-USk1p5gBO5BwBzy9iQmWlZdF8a_2hR37CnFJqSg7dDCW2XxYrGoAwC4gLhxtQ04_5FoLgF_FA6kBJwj-S_la0KuQX6qE1Q_maboQjag7LIhm9rDz7bkQTkwbGthc82ToD6AOh4EN/s1600/img5.PNG" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" height="297" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjDemc-USk1p5gBO5BwBzy9iQmWlZdF8a_2hR37CnFJqSg7dDCW2XxYrGoAwC4gLhxtQ04_5FoLgF_FA6kBJwj-S_la0KuQX6qE1Q_maboQjag7LIhm9rDz7bkQTkwbGthc82ToD6AOh4EN/s400/img5.PNG" width="400" /></a></div>
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<b><span style="font-size: 14.0pt; line-height: 115%;">Theory</span></b><span style="font-size: 14.0pt; line-height: 115%;">:<o:p></o:p></span></div>
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<span style="font-size: 14.0pt; line-height: 115%; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin;">1)<span style="font-size: 7pt; font-stretch: normal; font-variant-numeric: normal; line-height: normal;">    </span></span><span style="font-size: 14.0pt; line-height: 115%;">At the room temperature, the spore
wall which mostly contain keratin resist to dyes. But heating make the spore
wall more permeable to malachite green and allow it to bind to peptidoglycan
layer. <o:p></o:p></span></div>
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<span style="font-size: 14.0pt; line-height: 115%; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin;">2)<span style="font-size: 7pt; font-stretch: normal; font-variant-numeric: normal; line-height: normal;">    </span></span><span style="font-size: 14.0pt; line-height: 115%;">So both vegetative cell and endospore
are stained in green color.<o:p></o:p></span></div>
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<span style="font-size: 14.0pt; line-height: 115%; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin;">3)<span style="font-size: 7pt; font-stretch: normal; font-variant-numeric: normal; line-height: normal;">    </span></span><span style="font-size: 14.0pt; line-height: 115%;">After cooling, spore wall seals again
and trap malachite green inside the wall.<o:p></o:p></span></div>
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<span style="font-size: 14.0pt; line-height: 115%; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin;">4)<span style="font-size: 7pt; font-stretch: normal; font-variant-numeric: normal; line-height: normal;">    </span></span><span style="font-size: 14.0pt; line-height: 115%;">At the decolonization step, malachite
green rinse only forms the vegetative cells. <o:p></o:p></span></div>
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<span style="font-size: 14.0pt; line-height: 115%; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin;">5)<span style="font-size: 7pt; font-stretch: normal; font-variant-numeric: normal; line-height: normal;">    </span></span><span style="font-size: 14.0pt; line-height: 115%;">Counter stain, safranin I only able
to stain the vegetative cell because stain is unable to penetrate the spore
wall due to the less permeability. <o:p></o:p></span></div>
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Endospore staining

Endospore is domanet strucure which have no or less metabolic activities. Bactiers form endospre when environemntal conditions are unfavo...

Endospore staining

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