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65<br />
Alter 48 houls it€ nrmb€r ofcclls ofrlav@ werc counGd,n rhe inoculum<br />
mcdium following Brced.s Smear Method (Collin d 4/., l9E9). .Ih. iollowing sreps<br />
]vcrc rollorved.<br />
l) Loop of 4 didmeter (one toopfull=0.0tmt)<br />
'tun<br />
w6 prepared and dipped in<br />
2) Tte loop fu culrure waj sp€ad on mioos@pjc alide in a.e of tcm by first<br />
nrakinS rht d€s on nide rhm metinS it *irh mdker<br />
3) Slide wd air dried (n.n fixcd on flame.<br />
4) Staining was ptrfomed using Cnam.s st in, $e lta,mng ruft of which was as<br />
a) Crysralviolet 3 minul€s<br />
b) C.ah.s iodine 3 minur€s<br />
c) Acid alcohal washing<br />
d) Saf.ainine 5_r0 seconds<br />
5) The sljde so lrepeed w6 examined under l00X oil te!s.<br />
6) The microbial counr was pe.formed in five microscoprc lietds, to catculale rh€<br />
average mic.obiat count per field.<br />
?) Calcularions werc done as follows<br />
whe.e,<br />
Tobtnumber oforgeisnll,/mt - Ave.ae. No. ofo.aaaisms r M.f x pilurion<br />
o-ol<br />
M f= Micccopic fador^o or micrcscopic fi.los per cn.. which w.s<br />
bY<br />
1,,{.F "tt- '<br />
A =l0ru (mearx Icrn? ale, is equal to l0rmicrons)<br />
a = AEa, which is carculared by t',
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