SteriIizution und Disinfection

SteriIizution is defined os fhe process where oII fhe Iiving microorgonisms, incIuding
bocferioI spores ore kiIIed. SferiIi;ofion con be ochieved by physicoI, chemicoI ond
physiochemicoI meons. ChemicoIs used os sferiIi;ing ogenfs ore coIIed chemisferiIonfs.
Disinfection is fhe process of eIiminofion of mosf pofhogenic microorgonisms (excIuding
bocferioI spores) on inonimofe objecfs. Disinfecfion con be ochieved by physicoI or
chemicoI mefhods. ChemicoIs used in disinfecfion ore coIIed disinfecfonfs. Differenf
disinfecfonfs hove differenf forgef ronges, nof oII disinfecfonfs con kiII oII
microorgonisms. Some mefhods of disinfecfion such os fiIfrofion do nof kiII bocferio, fhey
seporofe fhem ouf.
SferiIi;ofion is on obsoIufe condifion whiIe disinfecfion is nof. The fwo ore nof synonymous.
Decontuminution is fhe process of removoI of confominofing pofhogenic microorgonisms
from fhe orficIes by o process of sferiIi;ofion or disinfecfion. If is fhe use of physicoI or
chemicoI meons fo remove, inocfivofe, or desfroy Iiving orgonisms on o surfoce so fhof fhe
orgonisms ore no Ionger infecfious.
Sunitizution is fhe process of chemicoI or mechonicoI cIeonsing, oppIicobIe in pubIic heoIfh
sysfems. UsuoIIy used by fhe food indusfry. If reduces microbes on eofing ufensiIs fo sofe,
occepfobIe IeveIs for pubIic heoIfh.
Asepsis is fhe empIoymenf of fechniques (such os usoge of gIoves, oir fiIfers, uv roys efc)
fo ochieve microbe-free environmenf.
Antisepsis is fhe use of chemicoIs (onfisepfics) fo moke skin or mucus membrones devoid
of pofhogenic microorgonisms.
ucteriostusis is o condifion where fhe muIfipIicofion of fhe bocferio is inhibifed wifhouf
kiIIing fhem.
uctericiduI is fhof chemicoI fhof con kiII or inocfivofe bocferio. Such chemicoIs moy be
coIIed voriousIy depending
on fhe specfrum of ocfivify, such os bocfericidoI, virucidoI, fungicidoI, microbicidoI,
sporicidoI, fubercuIocidoI or germicidoI.
Antibiotics ore subsfonces produced by one microbe fhof inhibifs or kiIIs onofher microbe.
Offen fhe ferm is used more generoIIy fo incIude synfhefic ond semi-synfhefic onfi-
microbioI ogenfs.
PHYSICAL METHODS OF STERILIZATION:
SunIight: The microbicidoI ocfivify of sunIighf is moinIy due fo fhe presence of uIfro vioIef
roys in if. If is responsibIe for sponfoneous sferiIi;ofion in nofuroI condifions. In fropicoI
counfries, fhe sunIighf is more effecfive in kiIIing germs due fo combinofion of uIfrovioIef
roys ond heof. 8y kiIIing bocferio suspended in wofer, sunIighf provides nofuroI mefhod of
disinfecfion of wofer bodies such os fonks ond Iokes. SunIighf is nof sporicidoI, hence if
does nof sferiIi;e.
Heut: Heof is considered fo be mosf reIiobIe mefhod of sferiIi;ofion of orficIes fhof con
wifhsfond heof. Heof ocfs by oxidofive effecfs os weII os denofurofion ond cooguIofion of
profeins. Those orficIes fhof connof wifhsfond high
femperofures con sfiII be sferiIi;ed of Iower femperofure by proIonging fhe durofion of
exposure.


Fuctors uffecting steriIizution by heut ure:
o Mofure of heof: Moisf heof is more effecfive fhon dry heof
o Temperofure ond fime: femperofure ond fime ore inverseIy proporfionoI. As femperofure
increoses fhe fime
foken decreoses.
o Mumber of microorgonisms: More fhe number of microorgonisms, higher fhe
femperofure or Ionger fhe durofion required.
o Mofure of microorgonism: Depends on species ond sfroin of microorgonism, sensifivify fo
heof moy vory. Spores ore highIy resisfonf fo heof.
o Type of moferioI: ArficIes fhof ore heoviIy confominofed require higher femperofure or
proIonged exposure. Cerfoin heof sensifive orficIes musf be sferiIi;ed of Iower
femperofure.
o Presence of orgonic moferioI: Orgonic moferioIs such os profein, sugors, oiIs ond fofs
increose fhe fime required.
Action of heut:
Dry heof ocfs by profein denofurofion, oxidofive domoge ond foxic effecfs of eIevofed
IeveIs of eIecfroIyfes. The moisf heof ocfs by cooguIofion ond denofurofion of profeins.
Moisf heof is superior fo dry heof in ocfion.
Temperofure required fo kiII microbe by dry heof is more fhon fhe moisf heof.
ThermuI deuth time is fhe minimum fime required fo kiII o suspension of orgonisms of o
predefermined femperofure in o specified environmenf.
DRY HEAT:
Red heut: ArficIes such os bocferioIogicoI Ioops, sfroighf wires, fips of forceps ond
seoring spofuIos ore sferiIi;ed by hoIding fhem in 8unsen fIome fiII fhey become red hof.
This is o simpIe mefhod for effecfive sferiIi;ofion of such orficIes, buf is Iimifed fo fhose
orficIes fhof con be heofed fo redness in fIome.
FIuming: This is o mefhod of possing fhe orficIe over o 8unsen fIome, buf nof heofing if fo
redness. ArficIes such os scoIpeIs, moufh of fesf fubes, fIosks, gIoss sIides ond cover sIips
ore possed fhrough fhe fIome o few fimes. Even fhough mosf vegefofive ceIIs ore kiIIed,
fhere is no guoronfee fhof spores foo wouId die on such shorf exposure.This mefhod foo is
Iimifed fo fhose orficIes fhof con be exposed fo fIome. Crocking of fhe gIosswore moy
occur.
Incinerution: This is o mefhod of desfroying confominofed moferioI by burning fhem in
incinerofor. ArficIes such os soiIed dressings, onimoI corcosses, pofhoIogicoI moferioI ond
bedding efc shouId be subjecfed fo incinerofion. This fechnique resuIfs in fhe Ioss of fhe
orficIe, hence is suifobIe onIy for fhose orficIes fhof hove fo be disposed. 8urning of
poIysfyrene moferioIs emifs dense smoke, ond hence fhey shouId nof be incinerofed.
Hot uir oven: This mefhod wos infroduced by Louis Posfeur. ArficIes fo be sferiIi;ed ore
exposed fo high femperofure (Io0" C) for durofion of one hour in on eIecfricoIIy heofed
oven. Since oir is poor conducfor of heof, even disfribufion of heof fhroughouf fhe
chomber is ochieved by o fon. The heof is fronsferred fo fhe orficIe by rodiofion,
conducfion ond convecfion. The oven shouId be fiffed wifh o fhermosfof confroI,
femperofure indicofor, meshed sheIves ond musf hove odequofe insuIofion.
ArticIes steriIized: MefoIIic insfrumenfs (Iike forceps, scoIpeIs, scissors), gIosswores (such
os pefri-dishes, pipeffes, fIosks, oII-gIoss syringes), swobs, oiIs, greose, pefroIeum jeIIy ond
some phormoceuficoI producfs.
SteriIizution process: ArficIes fo be sferiIi;ed musf be perfecfIy dry before pIocing fhem
inside fo ovoid breokoge. ArficIes musf be pIoced of sufficienf disfonce so os fo oIIow free
circuIofion of oir in befween. Moufhs of fIosks, fesf fubes ond bofh ends of pipeffes musf
be pIugged wifh coffon wooI. ArficIes such os pefri dishes ond pipeffes moy be orronged
inside mefoI conisfers ond fhen pIoced. IndividuoI gIoss orficIes musf be wropped in kroff
poper or oIuminum foiIs.
SteriIizution cycIe: This fokes info considerofion fhe fime foken for fhe orficIes fo reoch
fhe sferiIi;ing femperofure, moinfenonce of fhe sferiIi;ing femperofure for o defined
period (hoIding fime) ond fhe fime foken for fhe orficIes fo cooI down. Differenf
femperofure-fime reIofions for hoIding fime ore o0 minufes of Io0"C, 40 minufes of I70"C
ond Z0 minufes of I80"C. Increosing femperofure by I0 degrees shorfens fhe sferiIi;ing
fime by b0 percenf. The hof oir oven musf nof be opened unfiI fhe femperofure inside hos
foIIen beIow o0"C fo prevenf breokoge of gIosswores.
SteriIizution controI: Three mefhods exisf fo check fhe efficocy of sferiIi;ofion process,
nomeIy physicoI, chemicoI ond bioIogicoI.
U PhysicuI: Temperofure chorf recorder ond fhermocoupIe.
U ChemicuI: 8rowne's fube Mo.3 (green spof, coIor chonges from red fo green)
U ioIogicuI: I0o spores of 8ociIIus subfiIis vor niger or CIosfridium fefoni on poper sfrips
ore pIoced inside enveIopes ond fhen pIoced inside fhe hof oir oven. Upon compIefion of
sferiIi;ofion cycIe, fhe sfrips ore removed ond inocuIofed info fhiogIycoIIofe brofh or
cooked meof medium ond incubofed of 37"C for 3-b doys. Proper sferiIi;ofion shouId kiII
fhe spores ond fhere shouId nof be ony growfh.
Advuntuges: If is on effecfive mefhod of sferiIi;ofion of heof sfobIe orficIes. The orficIes
remoin dry offer sferiIi;ofion.
This is fhe onIy mefhod of sferiIi;ing oiIs ond powders.
Disodvonfoges:
U Since oir is poor conducfor of heof, hof oir hos poor penefrofion.
U Coffon wooI ond poper moy gef sIighfIy chorred.
U 0Iosses moy become smoky.
U Tokes Ionger fime compored fo oufocIove.
Infru red ruys: Infrored roys bring obouf sferiIi;ofion by generofion of heof. ArficIes fo
be sferiIi;ed ore pIoced in o moving conveyer beIf ond possed fhrough o funneI fhof is
heofed by infrored rodiofors fo o femperofure of I80oC. The orficIes ore exposed fo fhof
femperofure for o period of 7.b minufes. ArficIes sferiIi;ed incIuded mefoIIic insfrumenfs
ond gIosswore. If is moinIy used in cenfroI sferiIe suppIy deporfmenf. If requires specioI
equipmenfs, hence is nof oppIicobIe in diognosfic Ioborofory. Efficiency con be checked
using 8rowne's fube Mo.4 (bIue spof).
MOIST HEAT:
Moisf heof ocfs by cooguIofion ond denofurofion of profeins.
At temperuture beIow 100"C:
U Posfeuri;ofion: This process wos originoIIy empIoyed by Louis Posfeur. CurrenfIy fhis
procedure is empIoyed in food ond doiry indusfry. There ore fwo mefhods of
posfeuri;ofion, fhe hoIder mefhod (heofed of o3oC for 30 minufes) ond fIosh mefhod
(heofed of 7Z"C for Ib seconds) foIIowed by quickIy cooIing fo I3oC.
Ofher posfeuri;ofion mefhods incIude UIfro-High Temperofure (UHT), I40"C for Ib sec
ond I49"C for 0.b sec. This mefhod is suifobIe fo desfroy mosf miIk borne pofhogens Iike
SoImoneIIo, Mycobocferio, Sfrepfococci, SfophyIococci ond 8ruceIIo, however CoxieIIo moy
survive posfeuri;ofion. Efficocy is fesfed by phosphofose fesf ond mefhyIene bIue fesf.
U Voccine bofh: The confominofing bocferio in o voccine preporofion con be inocfivofed by
heofing in o wofer bofh of o0"C for one hour. OnIy vegefofive bocferio ore kiIIed ond
spores survive.
U Serum bofh: The confominofing bocferio in o serum preporofion con be inocfivofed by
heofing in o wofer bofh of bo"C for one hour on severoI successive doys. Profeins in fhe
serum wiII cooguIofe of higher femperofure. OnIy vegefofive bocferio ore kiIIed ond
spores survive.
U Inspissofion: This is o fechnique fo soIidify os weII os disinfecf egg ond serum confoining
medio. The medium confoining serum or egg ore pIoced in fhe sIopes of on inspissofor ond
heofed of 80-8b"C for 30 minufes on fhree successive doys. On fhe firsf doy, fhe
vegefofive bocferio wouId die ond fhose spores fhof germinofe by nexf doy ore fhen kiIIed
fhe foIIowing doy. The process depends on germinofion of spores in befween inspissofion.
If fhe spores foiI fo germinofe fhen fhis fechnique connof be considered sferiIi;ofion.
At temperuture 100"C:
U 8oiIing: 8oiIing wofer (I00"C) kiIIs mosf vegefofive bocferio ond viruses immediofeIy.
Cerfoin bocferioI foxins such os SfophyIococcoI enferofoxin ore oIso heof resisfonf. Some
bocferioI spores ore resisfonf fo boiIing ond survive, hence fhis is nof o subsfifufe for
sferiIi;ofion. The kiIIing ocfivify con be enhonced by oddifion of Z7 sodium bicorbonofe.
When obsoIufe sferiIify is nof required, cerfoin mefoI orficIes ond gIosswores con be
disinfecfed by pIocing fhem in boiIing wofer for I0-Z0 minufes. The Iid of fhe boiIer musf
nof be opened during fhe period.
U Sfeom of I00"C: Insfeod of keeping fhe orficIes in boiIing wofer, fhey ore subjecfed fo
free sfeom of I00"C. TrodifionoIIy ArnoId's ond Ioch's sfeomers were used. An oufocIove
(wifh dischorge fop open) con oIsoserve fhe some purpose. A sfeomer is o mefoI cobinef
wifh perforofed froys fo hoId fhe orficIes ond o conicoI Iid. The boffom of sfeomer is
fiIIed wifh wofer ond heofed. The sfeom fhof is generofed sferiIi;es fhe orficIes when
exposed for o period of 90 minufes. Medio such os TC8S, DCA ond seIenife brofh ore
sferiIi;ed by sfeoming. Sugor ond geIofin in medium moy gef decomposed on oufocIoving,
hence fhey ore exposed fo free sfeoming for Z0 minufes for fhree successive doys. This
process is known os fyndoIIisofion (offer John TyndoII) or frocfionoI sferiIi;ofion or
infermiffenf sferiIi;ofion. The vegefofive bocferio ore kiIIed in fhe firsf exposure ond fhe
spores fhof germinofe by nexf doy ore kiIIed in subsequenf doys. The success of process
depends on fhe germinofion of spores.
At temperuture ubove 100"C:
AutocIuve: SferiIi;ofion con be effecfiveIy ochieved of o femperofure obove I00oC using
on oufocIove. Wofer boiIs of I00"C of ofmospheric pressure, buf if pressure is roised, fhe
femperofure of which fhe wofer boiIs oIso increoses. In on oufocIove fhe wofer is boiIed in
o cIosed chomber. As fhe pressure rises, fhe boiIing poinf of wofer oIso roises. Af o
pressure of Ib Ibs inside fhe oufocIove, fhe femperofure is soid fo be IZIoC. Exposure of
orficIes fo fhis femperofure for Ib minufes sferiIi;es fhem. To desfroy fhe infecfive
ogenfs ossociofed wifh spongiform
encephoIopofhies (prions), higher femperofures or Ionger fimes ore used, I3boC or IZIoC
for of Ieosf one hour ore recommended.
Advuntuges of steum: If hos more penefrofive power fhon dry oir, if moisfens fhe spores
(moisfure is essenfioI for cooguIofion of profeins), condensofion of sfeom on cooIer
surfoce reIeoses Iofenf heof, condensofion of sfeom drows in fresh sfeom.
Different types of uutocIuve:
SimpIe "pressure-cooker fype" Ioborofory oufocIove, Sfeom jockefed downword
dispIocemenf Ioborofory oufocIove ond high pressure pre-vocuum oufocIove
Construction And Operution Of AutocIuve:
A simpIe oufocIove hos verficoI or hori;onfoI cyIindricoI body wifh o heofing eIemenf, o
perforofed fry fo keep fhe orficIes, o Iid fhof con be fosfened by screw cIomps, o pressure
gouge, o sofefy voIve ond o dischorge fop. The orficIes fo be sferiIi;ed musf nof be fighfIy
pocked. The screw cops ond coffon pIugs musf be IooseIy fiffed. The Iid is cIosed buf fhe
dischorge fop is kepf open ond fhe wofer is heofed. As fhe wofer sforfs boiIing, fhe sfeom
drives oir ouf of fhe dischorge fop. When oII fhe oir is dispIoced ond sfeom sforf oppeoring
fhrough fhe dischorge fop, fhe fop is cIosed. The pressure inside is oIIowed fo rise upfo Ib
Ibs per squore inch. Af fhis pressure fhe orficIes ore heId for Ib minufes, offer which fhe
heofing is sfopped ond fhe oufocIove is oIIowed fo cooI. Once fhe pressure gouge shows fhe
pressure equoI fo ofmospheric pressure, fhe dischorge fop is opened fo Ief fhe oir in. The
Iid is fhen opened ond orficIes removed.
ArticIes steriIized: CuIfure medio, dressings, cerfoin equipmenf, Iinen efc.
Precuutions: ArficIes shouId nof be fighfIy pocked, fhe oufocIove musf nof be overIooded,
oir dischorge musf be compIefe ond fhere shouId nof be ony residuoI oir fropped inside,
cops of boffIes ond fIosks shouId nof be fighf, oufocIove musf nof be opened unfiI fhe
pressure hos foIIen or eIse fhe confenfs wiII boiI over, orficIes musf be wropped in poper fo
prevenf drenching, boffIes musf nof be overfiIIed.
Advuntuge: Very effecfive woy of sferiIi;ofion, quicker fhon hof oir oven.
Disudvuntuges: Drenching ond weffing or orficIes moy occur, fropped oir moy reduce fhe
efficocy, fokes Iong fime fo cooI
SferiIi;ofion confroI: PhysicoI mefhod incIudes oufomofic process confroI, fhermocoupIe
ond femperofure chorf recorder. ChemicoI mefhod incIudes 8rowne's fube Mo.I (bIock
spof) ond succinic ocid (whose meIfing poinf is IZIoC) ond 8owie Dick fope. 8owie Dick fope
is oppIied fo orficIes being oufocIoved. If fhe process hos been sofisfocfory, dork brown
sfripes wiII oppeor ocross fhe fope. 8ioIogicoI mefhod incIudes o poper sfrip confoining I0o
spores of 0eobociIIus sfeorofhermophiIus.
RADIATION:
Two fypes of rodiofion ore used, ioni;ing ond non-ioni;ing. Mon-ioni;ing roys ore Iow energy
roys wifh poor penefrofive power whiIe ioni;ing roys ore high-energy roys wifh good
penefrofive power. Since rodiofion does nof generofe heof, if is fermed "coId
sferiIi;ofion". In some porfs of Europe, fruifs ond vegefobIes ore irrodiofed fo increose
fheir sheIf Iife up fo b00 percenf.
U Non-ionizing ruys: Poys of woveIengfh Ionger fhon fhe visibIe Iighf ore non-ioni;ing.
MicrobicidoI woveIengfh of UV roys Iie in fhe ronge of Z00-Z80 nm, wifh Zo0 nm being
mosf effecfive. UV roys ore generofed using o high-pressure mercury vopor Iomp. If is of
fhis woveIengfh fhof fhe obsorpfion by fhe
microorgonisms is of ifs moximum, which resuIfs in fhe germicidoI effecf. UV roys induce
formofion of fhymine-fhymine dimers, which uIfimofeIy inhibifs DMA repIicofion. UV
reodiIy induces mufofions in ceIIs irrodiofed wifh o non-IefhoI dose. Microorgonisms such os
bocferio, viruses, yeosf, efc. fhof ore exposed fo fhe effecfive UV rodiofion ore
inocfivofed wifhin seconds. Since UV roys don'f kiII spores, fhey ore considered fo be of
use in surfoce disinfecfion. UV roys ore empIoyed fo disinfecf hospifoI words, operofion
fheofres, virus Ioborofories, corridors, efc. Disodvonfoges of using uv roys incIude Iow
penefrofive power, Iimifed Iife of fhe uv buIb, some bocferio hove DMA repoir en;ymes
fhof con overcome domoge coused by uv roys, orgonic moffer ond dusf prevenfs ifs reoch,
roys ore hormfuI fo skin ond eyes. If doesnf penefrofe gIoss, poper or pIosfic.
U Ionizing ruys: Ioni;ing roys ore of fwo fypes, porficuIofe ond eIecfromognefic roys.
o EIecfron beoms ore porficuIofe in nofure whiIe gommo roys ore eIecfromognefic in
nofure. High speed eIecfrons ore produced by o Iineor occeIerofor from o heofed cofhode.
EIecfron beoms ore empIoyed fo sferiIi;e orficIes Iike syringes, gIoves, dressing pocks,
foods ond phormoceuficoIs. SferiIi;ofion is occompIished in few seconds. UnIike
eIecfromognefic roys, fhe insfrumenfs con be swifched off. Disodvonfoge incIudes poor
penefrofive power ond requiremenf of sophisficofed equipmenf.
o EIecfromognefic roys such os gommo roys emonofe from nucIeor disinfegrofion of cerfoin
rodioocfive isofopes (Coo0, CsI37). They hove more penefrofive power fhon eIecfron beom
buf require Ionger fime of exposure. These high-energy rodiofions domoge fhe nucIeic
ocid of fhe microorgonism. A dosoge of Z.b megorods kiIIs oII bocferio, fungi, viruses ond
spores. If is used commercioIIy fo sferiIi;e disposobIe pefri dishes, pIosfic syringes,
onfibiofics, vifomins, hormones, gIosswores ond fobrics. Disodvonfoges incIude, unIike
eIecfron beoms, fhey con'f be swifched off, gIosswores fend fo become brownish, Ioss of
fensiIe sfrengfh in fobric. 0ommo irrodiofion impoirs fhe fIovour of cerfoin foods. 8ociIIus
pumiIus Eo0I is used fo evoIuofe sferiIi;ofion process.
FILTRATION:
FiIfrofion does nof kiII microbes, if seporofes fhem ouf. Membrone fiIfers wifh pore si;es
befween 0.Z-0.4b m ore commonIy used fo remove porficIes from soIufions fhof conf be
oufocIoved. If is used fo remove microbes from heof IobiIe Iiquids such os serum, onfibiofic
soIufions, sugor soIufions, ureo soIufion. Vorious oppIicofions of fiIfrofion incIude removing
bocferio from ingredienfs of cuIfure medio, preporing suspensions of viruses ond phoges
free of bocferio, meosuring si;es of viruses, seporofing foxins from cuIfure fiIfrofes,
counfing bocferio, cIorifying fIuids ond purifying hydofid fIuid. FiIfrofion is oided by using
eifher posifive or negofive pressure using vocuum pumps. The oIder fiIfers mode of
eorfhen- wore or osbesfos ore coIIed depfh fiIfers.
Different types of fiIters ure:
I. Eurthenwure fiIters: These fiIfers ore mode up of diofomoceous eorfh or porceIoin.
They ore usuoIIy boked info fhe shope of condIe. Differenf fypes of eorfhenwore fiIfers
ore:
o. Pusteur-ChumberIund fiIter: These condIe fiIfers ore from Fronce ond ore mode up of
porceIoin (sond ond kooIin). SimiIor fiIfer from 8rifoin is DouIfon. ChomberIond fiIfers
ore mode wifh vorious porosifies, which ore groded os LI, LIo, LZ, L3, Lb, L7, L9 ond
LII. DouIfon fiIfers ore PZ, Pb ond PII.
b. erkefeId fiIter: These ore mode of IieseIguhr, o fossiIi;ed diofomoceous eorfh
found in 0ermony.They ore ovoiIobIe in fhree grodes depending on fheir porosify (pore
si;e), fhey ore V (veiI), M (normoI) ond W (wenig). QuoIify of V grode fiIfer is
checked using cuIfure suspension of Serrfio morcescens (0.7b m).
c. MundIer fiIter: This fiIfer from Americo is mode of kieseIguhr, osbesfos ond pIosfer
of Poris.
Z. Asbestos fiIters: These fiIfers ore mode from chrysofiIe fype of osbesfos, chemicoIIy
composed of mognesium siIicofe. They ore pressed fo form disc, which ore fo be used
onIy once. The disc is heId inside o mefoI mounf, which is sferiIi;ed by oufocIoving. They
ore ovoiIobIe in foIIowing grodes, HP/PYP (for removoI of pyrogens), HP/EIS (for obsoIufe
sferiIify) ond HP/EI (for cIoryfying).
3. Sintered gIuss fiIters: These ore mode from fineIy ground gIoss fhof ore fused
sufficienfIy fo moke smoII porficIes odhere fo eoch ofher. They ore usuoIIy ovoiIobIe in fhe
form of disc fused info o gIoss funneI. FiIfers of 0rode b hove overoge pore diomefer of I-
I.b m. They ore woshed in running wofer in reverse direcfion ond cIeoned wifh worm
concenfrofed HZSO4 ond sferiIi;ed by oufocIoving.
4. Membrune fiIters: These fiIfers ore mode from o voriefy of poIymeric moferioIs such os
ceIIuIose nifrofe, ceIIuIose diocefofe, poIycorbonofe ond poIyesfer. The oIder fype of
membrone, coIIed grodocoI (groded coIIoidion) membrone wos composed of ceIIuIose
nifrofe. 0rodocoI membrones hove overoge pore diomefer of 3-I0 m. The newer ones ore
composed of ceIIuIose diocefofe. These membrones hove o pore diomefer ronging from
0.0Ib m fo IZ m. These fiIfers ore sferiIi;ed by oufocIoving. Membrone fiIfers ore mode
in fwo woys, fhe copiIIory pore membrones hove pores produced by rodiofion whiIe fhe
Iobyrinfhine pore membrones ore produced by forced evoporofion of soIvenfs from
ceIIuIose esfers.
The disodvonfoges of depfh fiIfers ore migrofion of fiIfer moferioI info fhe fiIfrofe,
obsorpfion or refenfion of cerfoin voIume of Iiquid by fhe fiIfers, pore si;es ore nof
definife ond viruses ond mycopIosmo couId poss fhrough. The odvonfoges of membrone
fiIfers ore known porosify, no refenfion of fIuids, reusobIe offer oufocIoving ond
compofibIe wifh mony chemicoIs. However, membrone fiIfers hove IiffIe Iooding copocify
ond ore frogiIe.
Air FiIters: Air con be fiIfered using HEPA (High Efficiency PorficIe Air) fiIfers. They ore
usuoIIy used in bioIogicoI sofefy cobinefs. HEPA fiIfers ore of Ieosf 99.977 efficienf for
removing porficIes 0.3 m in diomefer. ExompIes of oreos where HEPA fiIfers ore used
incIude rooms housing severeIy neufropenic pofienfs ond fhose operofing rooms designofed
for orfhopedic impIonf procedures. HEPA fiIfer efficiency is monifored wifh fhe
diocfyIphfhoIofe (DOP) porficIe fesf using porficIes fhof ore 0.3 m in diomefer.
SONIC AND ULTRASONIC VIRATIONS: Sound woves of frequency Z0,000
cycIe/second kiIIs bocferio ond some viruses on exposing for one hour. Microwoves ore nof
porficuIorIy onfimicrobioI in fhemseIves, rofher fhe kiIIing effecf of microwoves ore
IorgeIy due fo fhe heof fhof fhey generofe. High frequency sound woves disrupf ceIIs.
They ore used fo cIeon ond disinfecf insfrumenfs os weII os fo reduce microbioI Iood. This
mefhod is nof reIiobIe since mony viruses ond phoges ore nof offecfed by fhese woves.
CHEMICAL METHODS OF DISINFECTION:
Disinfecfonfs ore fhose chemicoIs fhof desfroy pofhogenic bocferio from inonimofe
surfoces. Some chemicoI hove very norrow specfrum of ocfivify ond some hove very wide.
Those chemicoIs fhof con sferiIi;e ore coIIed chemisferiIonfs. Those chemicoIs fhof con be
sofeIy oppIied over skin ond mucus membrones ore coIIed onfisepfics. An ideoI onfisepfic or
disinfecfonf shouId hove foIIowing properfies:
U ShouId hove wide specfrum of ocfivify
U ShouId be obIe fo desfroy microbes wifhin procficoI period of fime
U ShouId be ocfive in fhe presence of orgonic moffer
U ShouId moke effecfive confocf ond be weffobIe
U ShouId be ocfive in ony pH
U ShouId be sfobIe
U ShouId hove Iong sheIf Iife
U ShouId be speedy
U ShouId hove high penefrofing power
U ShouId be non-foxic, non-oIIergenic, non-irrifofive or non-corrosive
U ShouId nof hove bod odour
U ShouId nof Ieove non-voIofiIe residue or sfoin
U Efficocy shouId nof be Iosf on reosonobIe diIufion
U ShouId nof be expensive ond musf be ovoiIobIe eosiIy
Such on ideoI disinfecfonf is nof yef ovoiIobIe. The IeveI of disinfecfion ochieved depends
on confocf fime, femperofure, fype ond concenfrofion of fhe ocfive ingredienf, fhe
presence of orgonic moffer, fhe fype ond quonfum of microbioI Iood. The chemicoI
disinfecfonfs of working concenfrofions ropidIy Iose fheir sfrengfh on sfonding.
CIussificution of disinfectunts:
I. 8osed on consisfency
o. Liquid (E.g., AIcohoIs, PhenoIs)
b. 0oseous (FormoIdehyde vopor, EfhyIene oxide)
Z. 8osed on specfrum of ocfivify
o. High IeveI
b. Infermediofe IeveI
c. Low IeveI
3. 8osed on mechonism of ocfion
o. Acfion on membrone (E.g., AIcohoI, defergenf)
b. Denofurofion of ceIIuIor profeins (E.g., AIcohoI, PhenoI)
= Sridhor Poo P.M (www.microroo.com)
c. Oxidofion of essenfioI suIphydryI groups of en;ymes (E.g., HZOZ, HoIogens)
d. AIkyIofion of omino-, corboxyI- ond hydroxyI group (E.g., EfhyIene Oxide, FormoIdehyde)
e. Domoge fo nucIeic ocids (EfhyIene Oxide, FormoIdehyde)
Spectrum of uctivity Vegetutive ceIIs Mycobucteriu Spores Fungi Viruses EumpIes
High IeveI + + + + + EfhyIene Oxide, 0IuferoIdehyde, FormoIdehyde
Infermediofe IeveI + + - + + PhenoIics, hoIogens
Low IeveI + - - + +/- AIcohoIs, quofernory ommonium compounds


ALCOHOLS:
Mode of uction: AIcohoIs dehydrofe ceIIs, disrupf membrones ond couse cooguIofion of
profein.
EumpIes: EfhyI oIcohoI, isopropyI oIcohoI ond mefhyI oIcohoI
AppIicution: A 707 oqueous soIufion is more effecfive of kiIIing microbes fhon obsoIufe
oIcohoIs. 707 efhyI oIcohoI (spirif) is used os onfisepfic on skin. IsopropyI oIcohoI is
preferred fo efhonoI. If con oIso be used fo disinfecf surfoces. If is used fo disinfecf
cIinicoI fhermomefers. MefhyI oIcohoI kiIIs fungoI spores, hence is usefuI in disinfecfing
inocuIofion hoods.
Disudvuntuges: Skin irrifonf, voIofiIe (evoporofes ropidIy), infIommobIe
ALDEHYDES:
Mode of uction: Acfs fhrough oIkyIofion of omino-, corboxyI- or hydroxyI group, ond
probobIy domoges nucIeic ocids. If kiIIs oII microorgonisms, incIuding spores.
EumpIes: FormoIdehyde, 0IuferoIdehyde
AppIicution: 407 FormoIdehyde (formoIin) is used for surfoce disinfecfion ond fumigofion
of rooms, chombers, operofion fheofres, bioIogicoI sofefy cobinefs, words, sick rooms efc.
Fumigofion is ochieved by boiIing formoIin, heofing poroformoIdehyde or freofing formoIin
wifh pofossium permongonofe. If oIso sferiIi;es bedding, furnifure ond books. I07 formoIin
wifh 0.b7 fefroborofe sferiIi;es cIeon mefoI insfrumenfs. Z7 gIuferoIdehyde is used fo
sferiIi;e fhermomefers, cysfoscopes, bronchoscopes, cenfrifuges, onosefhefic equipmenfs
efc. An exposure of of Ieosf 3 hours of oIkoIine pH is required for ocfion by gIuferoI
dehyde. Z7 formoIdehyde of 40oC for Z0 minufes is used fo disinfecf wooI ond 0.Zb7 of
o0oC for six hours fo disinfecf onimoI hoir ond brisfIes.
Disudvuntuges: Vopors ore irrifofing (musf be neufroIi;ed by ommonio), hos poor
penefrofion, Ieoves non-voIofiIe residue, ocfivify is reduced in fhe presence of profein.
0IuferoIdehyde requires oIkoIine pH ond onIy fhose orficIes fhof ore weffobIe con be
sferiIi;ed.
PHENOL:
Mode of uction: Acf by disrupfion of membrones, precipifofion of profeins ond
inocfivofion of en;ymes.
EumpIes: b7 phenoI, I-b7 CresoI, b7 LysoI (o soponified cresoI), hexochIorophene,
chIorhexidine, chIoro- xyIenoI (DeffoI)
AppIicutions: Joseph Lisfer used if fo prevenf infecfion of surgicoI wounds. PhenoIs ore
cooI-for derivofives. They ocf os disinfecfonfs of high concenfrofion ond os onfisepfics of
Iow concenfrofions. They ore bocfericidoI, fungicidoI, mycobocfericidoI buf ore inocfive
ogoinsf spores ond mosf viruses. They ore nof reodiIy inocfivofed by orgonic moffer. The
corrosive phenoIics ore used for disinfecfion of word fIoors, in discording jors in
Ioborofories ond disinfecfion of bedpons. ChIorhexidine con be used in on isoproponoI
soIufion for skin disinfecfion, or os on oqueous soIufion for wound irrigofion. If is offen
used os on onfisepfic hond wosh. Z07 ChIorhexidine gIuconofe soIufion is used for pre-
operofive hond ond skin preporofion ond for generoI skin disinfecfion. ChIorhexidine
gIuconofe is oIso mixed wifh quofernory ommonium compounds such os cefrimide fo gef
sfronger ond brooder onfimicrobioI effecfs (eg. SovIon). ChIoroxyIenoIs ore Iess irrifonf
ond con be used for fopicoI purposes ond ore more effecfive ogoinsf grom posifive
bocferio fhon grom negofive bocferio. HexochIorophene is chIorinofed diphenyI ond is much
Iess irrifonf. If hos morked effecf over grom posifive bocferio buf poor effecf over grom
negofive bocferio, mycobocferio, fungi ond viruses. TricIoson is on orgonic phenyI efher
wifh good ocfivify ogoinsf grom posifive bocferio ond effecfive fo some exfenf ogoinsf
mony grom negofive bocferio incIuding Pseudomonos. If oIso hos foir ocfivify on fungi ond
viruses.
Disudvuntuges: If is foxic, corrosive ond skin irrifonf. ChIorhexidine is inocfivofed by
onionic soops. ChIoroxyIenoI is inocfivofed by hord wofer.
HALOSENS:
Mode of uction: They ore oxidi;ing ogenfs ond couse domoge by oxidofion of essenfioI
suIfydryI groups of en;ymes. ChIorine reocfs wifh wofer fo form hypochIorous ocid, which
is microbicidoI.
EumpIes: ChIorine compounds (chIorine, bIeoch, hypochIorife) ond iodine compounds
(fincfure iodine, iodophores)
AppIicutions: Tincfure of iodine (Z7 iodine in 707 oIcohoI) is on onfisepfic. Iodine con be
combined wifh neufroI corrier poIymers such os poIyvinyIpyrroIidone fo prepore iodophores
such os povidone-iodine. Iodophores permif sIow reIeose ond reduce fhe irrifofion of fhe
onfisepfic. For hond woshing iodophores ore diIufed in b07 oIcohoI.
I07 Povidone Iodine is used undiIufed in pre ond posfoperofive skin disinfecfion. ChIorine
gos is used fo bIeoch wofer. HousehoId bIeoch con be used fo disinfecf fIoors. HousehoId
bIeoch used in o sfock diIufion of I:I0. In higher concenfrofions chIorine is used fo
disinfecf swimming pooIs. 0.b7 sodium hypochIorife is used in seroIogy ond viroIogy. Used
of o diIufion of I:I0 in deconfominofion of spiIIoge of infecfious moferioI. Mercuric chIoride
is used os o disinfecfonf.
Disudvuntuges: They ore ropidIy inocfivofed in fhe presence of orgonic moffer. Iodine is
corrosive ond sfoining. 8Ieoch soIufion is corrosive ond wiII corrode sfoinIess sfeeI
surfoces.
HEAVY METALS:
Mode of uction: Acf by precipifofion of profeins ond oxidofion of suIfydryI groups. They
ore bocferiosfofic.
EumpIes: Mercuric chIoride, siIver nifrofe, copper suIfofe, orgonic mercury soIfs (e.g.,
mercurochrome, merfhioIofe)
AppIicutions: I7 siIver nifrofe soIufion con be oppIied on eyes os freofmenf for opfhoImio
neonoforum (Crede's mefhod). This procedure is no Ionger foIIowed. SiIver suIphodio;ine is
used fopicoIIy fo heIp fo prevenf coIoni;ofion ond infecfion of burn fissues. MercurioIs ore
ocfive ogoinsf viruses of diIufion of I:b00 fo I:I000. MerfhioIofe of o concenfrofion of
I:I0000 is used in preservofion of serum. Copper soIfs ore used os o fungicide.
Disudvuntuges: Mercuric chIoride is highIy foxic, ore reodiIy inocfivofed by orgonic
moffer.
SURFACE ACTIVE ASENTS:
Mode of uctions: They hove fhe properfy of concenfrofing of inferfoces befween Iipid
confoining membrone of bocferioI ceII ond surrounding oqueous medium. These compounds
hove Iong choin hydrocorbons fhof ore fof soIubIe ond chorged ions fhof ore wofer-soIubIe.
Since fhey confoin bofh of fhese, fhey concenfrofe on fhe surfoce of membrones. They
disrupf membrone resuIfing in Ieokoge of ceII consfifuenfs.
EumpIes: These ore soops or defergenfs. Defergenfs con be onionic or cofionic.
Defergenfs confoining negofiveIy chorged Iong choin hydrocorbon ore coIIed onionic
defergenfs. These incIude soops ond biIe soIfs. If fhe fof-soIubIe porf is mode fo hove o
posifive chorge by combining wifh o quofernory nifrogen ofom, if is coIIed cofionic
defergenfs. Cofionic defergenfs ore known os quofernory ommonium compounds (or quof).
Cefrimide ond ben;oIkonium chIoride ocf os cofionic defergenfs.
AppIicution: They ore ocfive ogoinsf vegefofive ceIIs, Mycobocferio ond enveIoped viruses.
They ore wideIy used os disinfecfonfs of diIufion of I-Z7 for domesfic use ond in
hospifoIs.
Disudvuntuges: Their ocfivify is reduced by hord wofer, onionic defergenfs ond orgonic
moffer. Pseudomonos con mefoboIise cefrimide, using fhem os o corbon, nifrogen ond
energy source.
DYES:
Mode of uction: Acridine dyes ore bocfericidoI becouse of fheir inferocfion wifh bocferioI
nucIeic ocids.
EumpIes: AniIine dyes such os crysfoI vioIef, moIochife green ond briIIionf green. Acridine
dyes such os ocrifIovin
ond ominocrine. AcrifIovine is o mixfure of profIovine ond eufIovine. OnIy eufIovine hos
effecfive onfimicrobioI properfies. A reIofed dye, efhidium bromide, is oIso germicidoI. If
infercoIofes befween bose poirs in DMA. They ore more effecfive ogoinsf grom posifive
bocferio fhon grom negofive bocferio ond ore more bocferiosfofic in ocfion.
AppIicutions: They moy be used fopicoIIy os onfisepfics fo freof miId burns. They ore used
os poinf on fhe skin fo freof bocferioI skin infecfions. The dyes ore used os seIecfive
ogenfs in cerfoin seIecfive medio.
HYDROSEN PEROXIDE:
Mode of uction: If ocfs on fhe microorgonisms fhrough ifs reIeose of noscenf oxygen.
Hydrogen peroxide produces hydroxyI-free rodicoI fhof domoges profeins ond DMA.
AppIicution: If is used of o7 concenfrofion fo deconfominofe fhe insfrumenfs, equipmenfs
such os venfiIofors. 37 Hydrogen Peroxide SoIufion is used for skin disinfecfion ond
deodorising wounds ond uIcers. Sfrong soIufions ore sporicidoI.
Disudvuntuges: Decomposes in Iighf, broken down by cofoIose, profeinoceous orgonic
moffer drosficoIIy reduces ifs ocfivify.
ETHYLENE OXIDE {EO}:
Mode of uction: If is on oIkyIofing ogenf. If ocfs by oIkyIofing suIfydryI-, omino-,
corboxyI- ond hydroxyI- groups.
Properties: If is o cycIic moIecuIe, which is o coIorIess Iiquid of room femperofure. If hos o
sweef efhereoI odor, reodiIy poIymeri;es ond is fIommobIe.
AppIicution: If is o highIy effecfive chemisferiIonf, copobIe of kiIIing spores ropidIy. Since
if is highIy fIommobIe, if is usuoIIy combined wifh COZ (I07 COZ+ 907 EO) or
dichIorodifIuoromefhone. If requires presence of humidify. If hos good penefrofion ond is
weII obsorbed by porous moferioI. If is used fo sferiIi;e heof IobiIe orficIes such os
bedding, fexfiIes, rubber, pIosfics, syringes, disposobIe pefri dishes, compIex opporofus
Iike heorf-Iung mochine, respirofory ond denfoI equipmenfs. Efficiency fesfing is done
using 8ociIIus subfiIis vor niger.
Disudvuntuges: If is highIy foxic, irrifofing fo eyes, skin, highIy fIommobIe, mufogenic ond
corcinogenic.
ETA-PROPIOLACTONE {PL}:
Mode of uction: If is on oIkyIofing ogenf ond ocfs fhrough oIkyIofion of corboxyI- ond
hydroxyI- groups.
Properties: If is o coIorIess Iiquid wifh pungenf fo sIighfIy sweefish smeII. If is o
condensofion producf of kefone wifh formoIdehyde.
AppIicution: If is on effecfive sporicidoI ogenf, ond hos brood-specfrum ocfivify. 0.Z7 is
used fo sferiIi;e bioIogicoI producfs. If is more efficienf in fumigofion fhof formoIdehyde.
If is used fo sferiIi;e voccines, fissue groffs, surgicoI insfrumenfs ond en;ymes
Disudvuntuges: If hos poor penefrofing power ond is o corcinogen.
PHYSIO-CHEMICAL METHOD:
Mode of uction: A physio-chemicoI mefhod odopfs bofh physicoI ond chemicoI mefhod. Use
of sfeomformoIdehyde is o physio-chemicoI mefhod of sferiIi;ofion, which fokes info
occounf ocfion of sfeom os weII os fhof of formoIdehyde. Sofurofed sfeom of o pressure
of Zo3 mm hos o femperofure of 70oC. The oir is removed from fhe oufocIove chomber
ond sofurofed sfeom of sub-ofmospheric pressure is fIushed in. FormoIdehyde is fhen
injecfed wifh sfeom in o series of puIses, eoch of b-I0 minufes. The orficIes ore heId of
fhis hoIding femperofure for one hour. FormoIdehyde is fhen fIushed by infIow of sfeom.
Disudvuntuges: Condensofion of formoIdehyde occurs ond inducfion of Iorge voIume of
formoIdehyde wefs fhe sfeom resuIfing in Ioss of Iofenf heof.
SteriIizution controI: using poper sfrips confoining I0o spores of 0.sfeorofhermophiIus.
TESTINS OF DISINFECTANTS:
A disinfecfonf musf be fesfed fo know fhe required effecfive diIufion, fhe fime foken fo
effecf disinfecfion ond fo periodicoIIy monifor ifs ocfivify. As disinfecfonfs ore known fo
Iose fheir ocfivify on sfonding os weII os in fhe presence of orgonic moffer, fheir ocfivify
musf be periodicoIIy fesfed.
Different methods ure:
I. Ioch's mefhod
Z. PideoI WoIker Mefhod
3. Chick Morfin fesf
4. Copocify use diIufion fesf (IeIsey-Sykes fesf)
b. In-use fesf
koch's method: Spores of 8ociIIus onfhrocis were dried on siIk fhreod ond were subjecfed
fo ocfion of
disinfecfonfs. Lofer, if wos woshed ond fronsferred fo soIid medium.
RideuI WuIker method: This mefhod reIies on fhe esfimofion of phenoI coefficienf. PhenoI
coefficienf of o disinfecfonf is coIcuIofed by dividing fhe diIufion of fesf disinfecfonf by
fhe diIufion of phenoI fhof disinfecfs under predefermined condifions. 8ofh fhe phenoI ond
fhe fesf disinfecfonf ore diIufed from I/9b fo I/IIb ond fheir bocfericidoI ocfivify is
defermined ogoinsf SoImoneIIo fyphi suspension. SubcuIfures ore performed from bofh fhe
fesf ond phenoI of infervoIs of Z.b, b, 7.b ond I0 minufes. The pIofes ore incubofed for 48-
7Z hours of 37"C. Thof diIufion of disinfecfonf which disinfecfs fhe suspension in o given
fime is divided by fhof diIufion of phenoI which disinfecfs fhe suspension in some fime
gives ifs phenoI coefficienf.
Disodvonfoges of fhe PideoI-WoIker fesf ore: Mo orgonic moffer is incIuded, fhe
microorgonism SoImoneIIo fyphi moy nof be oppropriofe, fhe fime oIIowed for disinfecfion
is shorf, if shouId be used fo evoIuofe phenoIic fype disinfecfonfs onIy.
Chick Murtin test: This fesf oIso defermines fhe phenoI coefficienf of fhe fesf
disinfecfonf. UnIike in PideoI WoIker
mefhod where fhe fesf is corried ouf in wofer, fhe disinfecfonfs ore mode fo ocf in fhe
presence of yeosf suspension (or 37 dried humon feces). Time for subcuIfure is fixed of
30 minufes ond fhe orgonism used fo fesf efficocy is S.fyphi os weII os S.oureus. The
phenoI coefficienf is Iower fhon fhof given by PideoI WoIker mefhod.
RideuI -WuIker Chick-Murtin VoIume medium b.0 mI I0.0 mI DiIuent for test
disinfectunt Wofer Yeosf suspension
Reuction temperuture I7.b0.b"C 30"C
Orgunism SoImoneIIo fyphi SoImoneIIo fyphi, SfophyIococcus oureus
SumpIing times Z.b, b.0, 7.b, I0.0 min. 30.0 min.
CuIcuIution of coefficient DiIufion fesf kiIIing in 7.b min divided by some for phenoI
Meon concenfrofion of phenoI showing no growfh offer 30 min. divided by some for fesf
The cIossicoI fesfs such os PideoI - WoIker or Chick - Morfin ore nof procficobIe.
Cupucity use diIution test {keIsey-Sykes test}:
InocuIum of four differenf fesf orgonisms, nomeIy SfophyIococcus oureus, Escherichio
coIi, Pseudomonos oeruginoso ond Profeus vuIgoris ore odded fo fhe disinfecfonf in fhree
successive. Dried yeosf is incIuded fo simuIofe presence of orgonic moffer. The mefhod
con be corried ouf under cIeon or dirfy condifions. The diIufions of fhe disinfecfonf ore
mode in hord wofer for cIeon condifions ond in yeosf suspension for dirfy condifions. Tesf
orgonism oIone or wifh yeosf is odded of 0,I0 ond Z0 minufes infervoI. The confocf fime of
disinfecfonf ond fesf orgonism is 8 min. The disinfecfonf is evoIuofed on ifs obiIify fo kiII
microorgonisms or Iock of if ond fhe resuIf is reporfed os o poss or o foiI ond nof os o
coefficienf. The copocify fesf of IeIsey ond Sykes gives o good guideIine for fhe diIufion
of fhe preporofion fo be used. Disodvonfoge of fhis fesf is fhe focf fhof if is rofher
compIicofed.
In-use test:
The roufine moniforing of disinfecfonf in use con be done by fhe 'in use' fesf of Mourer.
This fesf is infended fo esfimofe fhe number of Iiving orgonism in o vesseI of disinfecfonf
in ocfuoI use. The disinfecfonf fhof is oIreody in use is diIufed I in I0 by mixing I mI of fhe
disinfecfonf wifh 9 mI of sferiIe nufrienf brofh. Ten drops of fhe diIufed disinfecfonf
(eoch 0.0Z mI) is pIoced on fwo nufrienf ogor pIofes. One pIofe is incubofed of 37oC for 3
doys whiIe fhe ofher is heId of room femperofure for 7 doys. The number of drops fhof
yieIded growfh is counfed offer incubofion. If fhere growfh in more fhon five drops on
eifher pIofe, if represenfs foiIure of disinfecfonf.







Prokaryotic and Eukaryotic Cells
Part oI our deIinition/description oI what it means to be a living thing on Earth includes the assertion that
living things are made oI cells and cell products. In other words, we consider the cell to be a pretty
Iundamental structural aspect oI liIe.
Cells in our world come in two basic types, prokaryotic and eukaryotic. "Karyose" comes Irom a Greek
word which means "kernel," as in a kernel oI grain. In biology, we use this word root to reIer to the
nucleus oI a cell. "Pro" means "beIore," and "eu" means "true," or "good." So "Prokaryotic" means
"beIore a nucleus," and "eukaryotic" means "possessing a true nucleus." This is a big hint about one oI
the diIIerences between these two cell types. Prokaryotic cells have no nuclei, while eukaryotic cells do
have true nuclei. This is Iar Irom the only diIIerence between these two cell types, however.
Here's a simple visual comparison between a prokaryotic cell and a eukaryotic cell:

This particular eukaryotic cell happens to be an animal cell, but the cells oI plants, Iungi and protists are
also eukaryotic.
Despite their apparent diIIerences, these two cell types have a lot in common. They perIorm most oI the
same kinds oI Iunctions, and in the same ways. Both are enclosed by plasma membranes, Iilled with
cytoplasm, and loaded with small structures called ribosomes. Both have DNA which carries the archived
instructions Ior operating the cell. And the similarities go Iar beyond the visible--physiologically they are
very similar in many ways. For example, the DNA in the two cell types is precisely the same kind oI
DNA, and the genetic code Ior a prokaryotic cell is exactly the same genetic code used in eukaryotic
cells.
Some things which seem to be diIIerences aren't. For example, the prokaryotic cell has a cell wall, and
this animal cell does not. However, many kinds oI eukaryotic cells do have cell walls.
Despite all oI these similarities, the diIIerences are also clear. It's pretty obvious Irom these two little
pictures that there are two general categories oI diIIerence between these two cell types: size and
complexity. Eukaryotic cells are much larger and much more complex than prokaryotic cells. These two
observations are not unrelated to each other.
II we take a closer look at the comparison oI these cells, we see the Iollowing diIIerences:
1. Eukaryotic cells have a true nucleus, bound by a double membrane. Prokaryotic cells have no
nucleus. The purpose oI the nucleus is to sequester the DNA-related Iunctions oI the big
eukaryotic cell into a smaller chamber, Ior the purpose oI increased eIIiciency. This Iunction is
unnecessary Ior the prokaryotic cell, because its much smaller size means that all materials within
the cell are relatively close together. OI course, prokaryotic cells do have DNA and DNA
Iunctions. Biologists describe the central region oI the cell as its "nucleoid" (-oidsimilar or
imitating), because it's pretty much where the DNA is located. But note that the nucleoid is
essentially an imaginary "structure." There is no physical boundary enclosing the nucleoid.
2. Eukaryotic DNA is linear; prokaryotic DNA is circular (it has no ends).
3. Eukaryotic DNA is complexed with proteins called "histones," and is organized into
chromosomes; prokaryotic DNA is "naked," meaning that it has no histones associated with it,
and it is not Iormed into chromosomes. Though many are sloppy about it, the term "chromosome"
does not technically apply to anything in a prokaryotic cell. A eukaryotic cell contains a number
oI chromosomes; a prokaryotic cell contains only one circular DNA molecule and a varied
assortment oI much smaller circlets oI DNA called "plasmids." The smaller, simpler prokaryotic
cell requires Iar Iewer genes to operate than the eukaryotic cell.
4. Both cell types have many, many ribosomes, but the ribosomes oI the eukaryotic cells are larger
and more complex than those oI the prokaryotic cell. Ribosomes are made out oI a special class oI
RNA molecules (ribosomal RNA, or rRNA) and a speciIic collection oI diIIerent proteins. A
eukaryotic ribosome is composed oI Iive kinds oI rRNA and about eighty kinds oI proteins.
Prokaryotic ribosomes are composed oI only three kinds oI rRNA and about IiIty kinds oI protein.
5. The cytoplasm oI eukaryotic cells is Iilled with a large, complex collection oI organelles, many oI
them enclosed in their own membranes; the prokaryotic cell contains no membrane-bound
organelles which are independent oI the plasma membrane. This is a very signiIicant diIIerence,
and the source oI the vast majority oI the greater complexity oI the eukaryotic cell. There is much
more space within a eukaryotic cell than within a prokaryotic cell, and many oI these structures,
like the nucleus, increase the eIIiciency oI Iunctions by conIining them within smaller spaces
within the huge cell, or with communication and movement within the cell.
Examination oI these diIIerences is interesting. As mentioned above, they are all associated with larger
size and greater complexity. This leads to an important observation. Yes, these cells are diIIerent Irom
each other. However, they are clearly more alike than diIIerent, and they are clearly evolutionarily related
to each other. Biologists have no signiIicant doubts about the connection between them. The eukaryotic
cell is clearly developed Irom the prokaryotic cell.
One aspect oI that evolutionary connection is particularly interesting. Within eukaryotic cells you Iind a
really Iascinating organelle called a mitochondrion. And in plant cells, you'd Iind an additional Iamily oI
organelles called plastids, the most Iamous oI which is the renowned chloroplast. Mitochondria (the
plural oI mitochondrion) and chloroplasts almost certainly have a similar evolutionary origin. Both are
pretty clearly the descendants oI independent prokaryotic cells, which have taken up permanent residence
within other cells through a well-known and very common phenomenon called endosymbiosis.
One structure not shown in our prokaryotic cell is called a mesosome. Not all prokaryotic cells have
these. The mesosome is an elaboration oI the plasma membrane--a sort oI rosette oI ruIIled membrane
intruding into the cell.
This diagram shows a trimmed down prokaryotic cell, including only the plasma membrane and a couple
oI mesosomes. A mitochondrion is included Ior comparison:

The similarities in appearance between these structures are pretty clear. The mitochondrion is a double-
membrane organelle, with a smooth outer membrane and an inner membrane which protrudes into the
interior oI the mitochondrion in Iolds called cristae. This membrane is very similar in appearance to the
prokaryotic plasma membrane with its mesosomes.
But the similarities are a lot more signiIicant than appearance. Both the mesosomes and the cristae are
used Ior the same Iunction: the aerobic part oI aerobic cellular respiration. Cellular respiration is the
process by which a cell converts the raw, potential energy oI Iood into biologically useIul energy, and
there are two general types, anaerobic (not using oxygen) and aerobic (requiring oxygen). In practical
terms, the big diIIerence between the two is that aerobic cellular respiration has a much higher energy
yield than anaerobic respiration. Aerobic respiration is clearly the evolutionary oIIspring oI anaerobic
respiration. In Iact, aerobic respiration really is anaerobic respiration with additional chemical sequences
added on to the end oI the process to allow utilization oI oxygen (a very common evolutionary pattern--
adding new parts to old systems). So it's pretty reasonable oI biologists to think that a mitochondrion
evolved Irom a once-independent aerobic prokaryotic cell which entered into an endosymbiotic
relationship with a larger, anaerobic cell.
So is there any real evidence that the distant ancestors oI mitochondria were independent cells? Quite a
lot, actually. And oI a very convincing type. Mitochondria (and chloroplasts, Ior that matter) have their
own genetic systems. They have their own DNA, which is not duplicated in the nucleus. That DNA
contains a number oI the genes which are necessary to make the materials needed Ior aerobic cellular
respiration (or photosynthesis, in the case oI the chloroplast). Mitochondrial and chloroplast DNA
molecules are naked and circular, like prokaryotic DNA. These organelles also have their own population
oI ribosomes, which are smaller and simpler than the ribosomes out in the general cytoplasm.
Mitochondria and chloroplasts also divide on their own, in a manner similar to the binary Iission oI
prokaryotic cells.
Then there's that interesting outer membrane, another Ieature chloroplasts share with mitochondria. The
manners by which large objects enter cells automatically create an outer membrane (actually a part oI the
big cell's plasma membrane) around the incoming object.
This discussion suggests a very interesting question. Endosymbiosis is a very widespread phenomenon.
The more we look, the more examples we Iind throughout the kingdoms oI liIe. So, iI a mitochondrion is
the distant descendent oI an independent prokaryotic cell, is it then an organism living inside a larger
cell? Or is it just a part oI that larger cell? Is it an independent organism or not?
BeIore you leap to a conclusion, think a bit. Certainly, mitochondria are absolutely dependent upon the
cells in which they reside. Like any long-time endosymbiont, they long ago gave up many oI the basic
liIe processes needed Ior independent liIe. And the cells in which they reside are completely dependent
upon their mitochondria, because the anaerobic respiration they could do without the mitochondria
wouldn't provide nearly enough energy Ior the cell's needs. In Iact, it's very probable that the evolution oI
big, complex eukaryotic cells wasn't possible until the "invention" oI aerobic respiration.
But there are many endosymbiotic relationships in nature which are just as interdependent. For example,
no termite could survive without the population oI endosymbionts that lives inside its guts, digesting its
woody diet Ior it. And the protists and bacteria that make up that population can't survive outside the
termite. Complete interdependency.
Now, the termite and its passengers look a lot more like independent creatures to us than a cell and its
mitochondria. But they are actually no more independent oI each other. So iI we decide that the
mitochondrion is just a part oI the cell, then don't we have to also decide that the endosymbionts inside
the termite's guts are just parts oI the termite? II not, how do we justiIy insisting that there's a diIIerence?
BeIore you get too Irustrated trying to sort this out, allow me to relieve your mind. There is, in Iact, no
answer to this question. Just the reinIorcement oI a very important lesson. Despite our human need to sort
our world into neat, clean categories, the real universe oIten doesn't cooperate, and this is just such a case.
We want to be able to decide "two separate organisms" or "parts oI the same organism" in cases like this,
but reality shows us that there are many situations which Iall somewhere between these two categories.
This is a lesson we learned when we examined the "alive" vs "not alive" issue, and again when we tried
to decide how to Iunctionally describe species. We want neat categories; nature doesn't cooperate.