"

INTERVI
EW QUESTI
ONS&ANSWERS"

Q.I
ntr
oducey
our
sel
f.
1.Tel
l t
her ecr
uit
erabouty
ourqual
i
ficat
ion
start
ingfrom yourr
ecenttot
he12th
education.
2.
Fami
l
ybackgr
ound
3.
Abouty
ourst
rengt
hs
Q.
Whati
sSt
eri
l
izat
ion?
>Theprocesswhi chdestroysal
ltheformsof
microbi
all
if
ei ncludi
ngFungus,Vi
ruses&v i
able
cel
lsofmicro-organi
smsal ongwiththei
r
sporesbyusingChemi calaswellasPhy si
cal
methodsiscalledsteri
l
izati
on.
Ther
ear
emaj
or2met
hodsofst
eri
l
izat
ionas,
1]
Phy
sical
met
hod-
a)
Heat
ing-
i
)Dryheatst
eri
li
zat
ion-
160°C-180°
Cfor1.
5hrs-
Forgl
asswares&Heatstablecompounds.
EX.
Hotai
rov
en
*Fl
aming-Al
socall
edInser
inat
ionmethodf
or
t
hester
il
izat
ionofNichr
omewi rel
oop.
i
i)
Moi stheatsteri
li
zati
on-121°C-134°Cunder15
l
bspr essurefor20mi n.Asthepr essure
i
ncreasest empratur
eincreases.Usedf or
St
eril
izati
onofGr owthmedi a(
liquid&soli
d)&
moistresistantmater
ials.
EX.
AUTOCLAVE(
Steam underpr
essur
e)
i
i
)Paseur
izat
ion
-
Descr
ibePaseur
izat
ionandi
tst
ypesi
fasked.
b)
Radi
ati
on-
kil
l
ingofmi
crobesbyusi
ng
U.
V.r
ays,
Gammar
ays
usedt
odest
roybact
eri
alDNA.
Usedforheatsensi
ti
veproduct
s&Fordr
y
phar
maceuticalpr
oduct
s.
c)Fi
l
trat
ion-
Compl
eteRemov
alofMi
crobesi
s
carr
iedout.

I
nSter
il
izat
ionbyFil
trati
on0.
22micr
on
di
ametercell
ulosef
il
teri
sused.
LAMI
NARAI
RFLOW-
Pr
inci
ple:
-Cont
ini
ouspar
all
elf
lowofst
eri
l
eai
r
i
spassedt
othewor
kingar
eat
hrought
hef
il
ter
s
.
*Descr i
bewor kingofLAF: -
Inal aminarfl
ow
hoodt heai rispassedt hroughaHEPA( High
Eff
iciencyPar t
icul
atesAir)fi
lterwhichremov es
all
airbornecont aminati
ont omai ntai
nsteri
l
e
conditi
ons.. ..Nowt hesteril
eairflowsintothe
working( fl
asking)ar eawherey oucandoal l
yourflaskingwor kwithoutri
skof
contami nation.
☆HEPAFI LTERS(Hi
gheff
iciencypart
icul
ateai
r)
-Removes99.97%ofcont
ami nantpar
ticl
es
aboutof0.
3um diamet
ersize.
☆ULPAFI
LTERS(
Ult
ra-
lowpar
ti
cul
ateai
r)
-Moreeff
ici
entt
hanHEPAfi
lt
erremov
es
99.999%ofcont
aminant
sof0.
12um diamet
er
size.
2]
Chemi
cal
met
hod-
Iti
ncl
udes,

*Gaseousmet
hod-
For
mal
dehy
de,
Ethy
leneoxi
de,
Met
hanol
etc.

Q.
Howwi
l
lyoudi
ff
renci
atet
heBact
eri
a?
-Descri
bePrinci
pleofGr
am st
aini
ng&i
ts
procedure.
Q.
Bact
eri
alspor
est
aini
ng.
-
Pri
nci
pleofSpor
eSt
aini
ng:
Adifferential
staini
ngtechnique(the
Schaeffer-Ful
tonmet hod)isusedt o
dist
inguishbet weenthevegetati
vecells
andt heendospor es.Aprimarystain
(malachitegreen)isusedt ostai
nt he
endospor es.

Q.
Fungal
spor
est
aini
ng.
-
Expl
ainTheLact
ophenol
Cot
tonBl
ue
st
aini
ng.
Pr
inci
ple
LPCBisastai
nusedf ormaki
ngsemi-
permanentmi
croscopicpr
epar
ati
onof
fungi
.
TheLPCBstai
nhasf
oll
owi
ngt
hree
component
s:
Phenol
:Ki
l
lsanyl
i
veor
gani
sm.
Lact
icaci
d:Pr
eser
vesf
ungal
str
uct
ures.
Cot
tonblue:
Stai
nst
hechiti
nandcel
l
ulose
oft
hefungal
cell
wal
lint
ensel
ybl
ue.

Q.
Giv
esomeexampl
esofGr
am +v
e&Gr
am
-
veBact
eri
a.
-
Gr
am Posi
ti
ve Gr
am Negat
ive
-
B.subt
il
is -
E.col
i
-
B.megat
eri
um -
Acet
obact
er
-
B.t
hur
ingenensi
s -
Campy
lobact
er
-
Clost
ri
dium bot
uli
nm -
Nit
robact
er
-
Act
inomy
ces -
Pseudomonas
-
Cor
nybact
eri
um -
Sal
monel
l
a
-
Lact
obaci
l
lus -
Vibr
io
-
Mycobact
eri
um t
b -
Rickt
tesi
a
St
aphy
lococcus -
Yer
sini
a
St
rept
ococcus -
Klebsi
ell
a
St
rept
omy
ces -
Shi
gel
l
a

Q.
Giv
esomeexampl
esofFungus.
>A.niger
,A.
fl
avus,
Candi
daaur eus,
Candida
al
bicans,
Tri
chodermaviri
dae,
Penici
li
um
cr
ysogenum,Penici
li
um notat
um,Fusari
um
Q.
Whati
sDi
sinf
ect
ion?
>Disinfect
ioni stheprocessofremovi
ngor
ki
lli
ngtheal l
thef orm ofmicr
obiall
i
fe
exceptbacterialsporesiscal
led
disi
nfecti
on.
Thi
spr ocessi
sgeneral
lyappl
iedforthe
di
sinfect
ionofsurf
acesofpharmaceuti
cal
i
nstruments,
Surf
acesofworkingareaetc.
I
nPharmaceuti
cal
smostl
y70%Isopropyl
al
cohol
(70%IPA)i
susedast
hedisi
nfect
ant.
Q.
Why70%I sopropyl
alcoholi
susedas
di
sinf
ect
antinpharmaceuti
cal
i
ndust
ri
es,
whynot100%?
I
so-pr
opylal
cohol(
IPA)i
swi del
yusedasa
di
si
nfect
antinphar
maceuticalcompani
es.I
PA
i
sveryef
fecti
veongram posit
iveaswel
las
gram negat ivebacter
ia.Mostoft hehand
disinfectantsavail
abl
eint hemar ketcontai
ns
IPAasanact i
vecomponentj ustbecauseofit
s
effectiveness.IPAisveryeffecti
veandkill
mi crobialcell
wi t
hinaf ewseconds.Butone
l
imi tati
onofI PAisthatiti
snotef fecti
ve
agai nstbacteri
alandfungal spores.
Ther
easonisbehi
ndt
hemodeofact
ionof70%
I
PAisthat>
TheBact er
ialcell
shav e
proteinsi ntheircellwal l
andwhent hisprotein
comesi ncont actwi ththe70%I PAdur i
ng
disinf
ect antapplicat i
on,coagulati
onofpr otei
ns
takespl acesi nwhi chdenat urat
ionofpr oteins
occur r
edandaf t
ert hatIPApenet rat
ei nthe
i
nt eri
oroft hecel lwhichcausel ysisordeat hof
thecel l
.Pr otei
ncoagul ati
onalsohappensi n
caseof100%I PAbutwi thveryfastrateand
becauseoft hisv eryfastpr ot
eincoagul at
ion
processdenat uredpr oteinformspr otecti
v e
l
ayeroutsideoft hecel l.Whent hishappens,
100%cannotpenet rat einsidet hecel landnot
abletokil
lthemi crobe.Mi croor ganisms
becomedor manti nthatcondi t
ions.Incaseof
70%I PAproteincoagul ationt akespl acewi t
ha
sl
owr ateandduet othi s70%I PAgetenough
ti
met openet r
atei nthecel l andki l
l t
he
microorgani
sm.Anot herf act oriscont acttime,
70%I PAtakesl ongert i
met oev aporatef r
om
anysurfacehencegetenoughcont actt i
meand
i
nt hi
smeant i
mei tshowi t sef fi
cacybuti n
caseof100%I PA, evapor at i
onwi llbev eryfast
,
contactti
mewi l
lbel essandi twillnotbeso
eff
ectiveagainstmi cr obes.That '
swhy70%I PA
solut
ionisusedasdi sinfect anti n
pharmaceuticalsindust r
ies.

Q.
Whatar
eant
isept
ics?
>Ant
isept
icsar
ethechemi
cal
subst
ances
whichar eusedindest
roy
ingdiseasecausi
ng
microorganisms(al
socal
ledpathogens)
ext
er nal
lyonwoundsorappli
edonski nsur
face
totreati
nfecti
on.
Ex.
Sani
ti
zer
Mostchemical agentscanbeusedasbot han
anti
septi
candadi sinfectant.Thepurposef or
whichiti
susedi sdeter minedbyi t
s
concentr
ati
on.Forexampl eHy drogenperoxide
6%soluti
oni susedforcl eansingwounds, whil
e
str
ongersoluti
ons(>30%)ar eusedi nindustry
asableachandox i
disingagent .
Iodineisusual
lyusedinanalcoholsolution
(call
edtinct
ureofiodi
ne)orasLugol'siodine
soluti
onasapr e-andpostoperat
iveantisepti
c.
Q.
Howwat
eranal
ysi
siscar
ri
edout?
-
Descri
beaboutMPNwhichi
sonlycar
ri
edof
f
orthedet
ecti
onofCol
i
formsint
heWat er.
-
AlsoMLT(
Micr
obi
all
i
mitt
est)i
scar
ri
edoutf
or
wat
eranal
ysi
s,alsoter
medasTAMC( Tot
al
Aer
obi
cMicrobialcount)orTVAC(
Tot
alVi
abl
e
Aer
obi
cCount)asperUSP.
Q.
Whati
sC.
F.U?
>Acolony-f
ormingunit(CFU)isauni tthati
s
usedtoestimatethenumberofv iablebacteri
a
orfungalcel
lsinasampl e.Col
onyf orming
unit
sareusedasameasur eoft
henumberof
micro-
organismspresentinoronsur faceofa
sample.
Todet erminethenumberofcolonyforming
unit
s, asampleispreparedandspreador
poureduni f
ormlyonasur f
aceofanagarpl at
e
andtheni ncubatedatsomesuit
able
temper at
ureforanumberofday s.Thecoloni
es
thatformsar ecounted.
(watchony
out
ube,
vi
deoofCFUbyShomu'
s
biol
ogy)

Q.
Howwi
l
lyoucal
cul
ateCFU?

Q.
Whati
sBi
obur
dent
est
ing?
>Bioburdentest
ingi
sper f
ormedt odet ermine
thetotalnumberofaerobi
cv iable
microorgani
smsi noronamedi caldevice,
contai
nerorcomponentaf tercompl et
ionofall
i
n-processstepspri
ortosteril
izat
ion.
Q.
WhatareSel
ecti
ve,
Dif
fer
ent
ial
,Enr
iched,
Nut
ri
entandMini
malmedia?
I
nmi
crobi
ology
,weusedi
ff
erentt
ypeofmedi
a
onrout
inebasi
s.Di
ffer
entmedi ahav
edif
fer
ent
pr
opert
iesandusedfordiff
erentt
ypeof
micr
oorgani
sms.Dif
ferentty
pesofmediaar
e
menti
onedbelow.

Sel
ect
iveMedi
a:
Selectivemedi aar et hosemedi awhi char e
usedt osuppor tgrowt hofonegr oupof
mi croorgani
smsbuti nhi bitthegr owt hofot her
groupofmi croor ganisms.Forexampl eincase
ofManni t
olsal tagarmedi a.Itcont ai
nsv ery
highamountofsal t(7.5%)whi chi nhi
bitthe
growt hofgr am negat i
v ebact eriaandi tis
selectiveforgr am posi tivebact eriali
ke
staphy l
ococcus.MacConkeyagarmedi aisalso
selectiveforgr am negat i
vebact eriaandot her
bact er
iawhi chf oundi nt hei ntestinaltr
ack.
MacConkeyagarcont ainsbi lesal t
sandcr y st
al
vi
ol etwhichi nhibitthegr owt hofgr am positive
bact er
ia.
Di
ff
erent
ial
Medi
a:
Diff
erent i
al mediaar ethosemedi awhi char e
usedt odi sti
nguishcl osel
yr elated
microor ganismsbasedont hei rmor phol ogyon
di
fferentagarmedi a.Forexampl eincaseof
Manni tolsaltagar ,itcontainsmanni tol and
phenol redaspHi ndi cat
orwhi chsuppor tt he
growthofmanni tol fermentingst aphy l
ococcus.
Acidpr oducedduet omanni tol f
erment ationis
detectedandduet opr esenceofphenol red
i
ndicatordy estaphy lococcusshowsy ellow
colouredcol oniesonmanni tol saltagarmedi a.
MacConkeyagari sal sodifferenti
al medi a
whichcont ainslactoseandi tsuppor tthe
growthofl actosef erment i
ngbact erialikeE. col
i.
MacConkeyAgarMedi
a
Manni
tol
Sal
tAgarMedi
a
So,
medi
acoul
dbebot
hsel
ect
iveaswel
las
di
ff
erent
ial
.
Enr
ichedmedi
a:
Thesemedi asupportthegrowt hofwi devar i
ety
ofmi croor
ganismsanddoesn' tinhi
bitthe
growt hofmi cr
oorganisms.Thesemedi a
containshighamountofnut r
ientsandmai nly
usedt oharvestthealltypesofmi cr
oor ganism
whichar epresentinthesampl e.Forexampl e
Soyabeancasei ndigestmedium i susedf or
enri
chmentofsampl es.
Nut
ri
entmedi
a:
Thesemediaarealsocall
edgeneralpur pose
media.Thesemediaarenonselecti
v eandt hey
contai
ngeneralnut
ri
entswhichrequiredforthe
growthofwiderangeofmicroor
ganisms.
Soyabeancasei
ndigestagarandNut rientagar
aretheexampleofthi
stypeofmedi a.
Mi
nimal
medi
a:
Mi
nimal
medi
aar
ethosemedi
awhi
chcont
ains
mini
mal nutr
ientsforthegr owthof
microorganisms.Thesemedi aaremainl
yused
forf
astidiousmi croorganisms.R2Amediai
san
exampleofmi nimal mediainwhichnut
ri
ent
s
arepresentinv erysmal lamount.
Medi
awi
thcomposi
ti
on:
1.
MacConkeyAgar
MacConkeyAgari srecommendedf orselecti
ve
i
solationofEscheri
chiacolif
rom
pharmaceut i
calproductsandisinaccordance
withharmonizedmet hodologyofBP.Itisalso
recommendedf orselect
ivei
solat
ionand
diff
erenti
ati
onoflactosefermentingand
l
actosenonf erment i
ngenteri
cbacteri
a.

Composi
ti
on*
*
Pept
ones(
meatandcasei
n)
Pancr
eat
icdi
gestofgel
ati
n
Lact
osemonohy
drat
e
Bi
l
esal
ts
Sodi
um chl
ori
de
Cr
yst
alv
iol
et
Neut
ral
red
Agar
pHaf
terst
eri
l
izat
ion(at25°
C)7.
1±0.
2

Theselectiveacti
onoft hismedium i
s
att
ribut
edt ocrystalv
ioletandbil
esalt
s,which
areinhi
bitor
yt omostspeci esofgram-posi
ti
ve
bacteri
a.Sodium chl
oridemaintai
nstheosmot i
c
balanceinthemedi um.
2.
Nut
ri
entAgarMedi
um
Nutri
entagarmedium i
susedasagener al
pur
posecultur
emedi um whichmaybeusedas
enr
ichedmedium byincorporat
ingbl
oodor
ot
herbi
ologi
calf
lui
dsi
naccor
dancewi
thI
ndi
an
Phar
macopoeia.
Composi
ti
on*
*
Pept
one
Meatext
ractB#
Sodi
um chl
ori
de
Agar
pHaf
terst
eri
l
izat
ion7.
3±0.
1
Q.
Whati
sBETt
est?
-I
tistheMicr
obiologi
cal
testwhi
chcarr
iedout
todetectt
hepresenceofBacter
ial
endotoxi
n
presenti
nthegivensample.
Thisendotoxi
ntesti
salsocall
edasLALTest -
becauseinthi
stestt
he'LimulusAmebocyte
Lysat
e'isusedwhichistheaqueousext
ractof
bl
oodcel l
sofHorseshoecrab.
I
tsi
sal
socal
l
edGel
Clotmet
hodbczi
nthi
sthe
sampl eist
akeninsmal l
tubesthenf ewdrops
ofLimulusamebocy t
eLy sat
eareadded&kept
forincubat
ionat37°C.Af
teri
ncubationifthe
sampl eshowsGel cl
ot,i
tist
heposit i
vetest
thatindi
catespr
esenceofendot oxi
n.
Endotoxi
nsarenothingbutt he
l
ippopoly
sacchari
despr esenti
nthecel
lwal
lof
Gram negat
ivebacteri
a.
LALtestist
heQuali
tat
ivetest&KTA(Kinet
ic
Turbi
dometri
cAssay)istheQuanti
tat
ivetestof
Endotoxi
n.

Q.
Whati
spy
rogent
est
ing?
-Thi
smet hodi
scarr
iedoutt
odetectt
he
presenceofpy
rogen.(
Amestest
)
Q.
Whati
sNor
mal
i
ty?
-I
tisdefi
nedastheNumberofmol
es
equiv
alentperl
i
tresol
uti
on.
Q.Envi
ronmental
Monitor
ingi
n
Pharmaceuti
cali
ndust
ry.
-
Itiscar
riedoutt
oanaylset
hepresenceof
Microbi
alloadpr
esenti
npharmaceuti
cal
i
ndustry
.
Ther
earemajorfol
l
owingmet
hodsof
Env.
monit
ori
nginphar
maceut
ical
indusr
tyas-
1.
Set
tl
epl
atemet
hod-90mm
2.
Cont
actpl
atemet
hod-
55mm
3.
Swabt
est
ing
4.
Airsampl
er(
Ainst
rument
)
5.
Per
sonnel
moni
tor
ing
ForE.
M.i
nphar mamostl
ySCDA(Soyabean
Caesi
nDigestAgar)i
susedf
orbot
hFungus
andBact
erial
growth.
Q.
Descr
ibeMi
croscope?
Q.Whati
sdi
ff
rencebetweensi
mpl
eand
compoundMicr
oscope?
-I
nsimplemicr
oscopet her
eissingl
elens,
whereasincompoundl i
ghtmicr
oscopethere
are3to5object
ivelenses,whi
chmeans
compoundmi croscopehasbettermagnif
ying
power.
Q.
Whati
sthet
otal
magni
fi
cat
ionof
compoundmi
croscope?
-10+40+100×10(
Eyepi
ece)
=1500X
Q.
Whoi
sthef
atherofMi
crobi
ology?
-
Antoni
ePhili
psv
anLeeuwenhoeki
sknownas
'
TheFatherofMi
crobi
ology
'.
Q.
Whoi
sfat
herofModer
nMi
crobi
ology?
-Loui
sPasteurr
egardedasthefat
herof
moder nmicr
obiol
ogy.Heconfi
rmedthe
previ
ousexperi
mentstodispr
ovespont
aneous
generati
on.
Q.
Descr
ibeaci
dfastst
aini
ng?
-htt
p://
www. sl
ideshare.net
/MMASSY/ aci
d-f
ast
-
stai
ning-pr
ocedure-f
or-stai
ning-
mycobact
eri
a?
Q.Enli
stthebiochemical
test
susedf
ort
he
bacteri
alident
if
icat
ion?
-
IMVI
C,MRVP,
Sugart
est
Q.
Whati
sther
oleofCondenseri
n
Mi
croscope?
-
condensergather
slightfrom themi croscope
l
i
ghtsour ceandconcentratesitint
oaconeof
l
i
ghtt hatil
l
uminat
est hespecimenwi t
huni f
orm
i
ntensit
yov ert
heenti
reviewfield.
Q.
IsMcConkey
sagarsel
ect
iveordi
ff
renci
al
?
Q.
Howwi l
lyoudi
ff
renci
ateEnt
erobact
eron
McConkey
sagar?
Q.
Whatar
ethet
ypesofE.
col
i?
-
Ent
erot
oxi
geni
cE.col
i(ETEC)
-
Ent
eropat
hogeni
cE.col
i(EPEC)
-
Ent
eroi
nvasi
veE.col
i(EI
EC)
-
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