Cul

ti
vat
ionofAnaer
obi
cbact
eri
a
Av
ari
ety ofmet
hods ar
e av
ail
abl
efort
he cul
tur
e ofanaer
obi
c
mi
croor
gani
smsi
nthel
abor
ator
y.

Anaer
obi
cmet
hodsar
edescr
ibedf
orachi
evi
nganaer
obi
osi
sont
hebasi
s
off
oll
owi
ngpr
oper
ti
es

1.Pr
oduct
ionofv
acuum

2.Di
spl
acementofoxy
genwi
thot
hergases

3.Absor
pti
onofoxy
genbychemi
cal
orbi
ologi
cal
met
hods

4.Absor
pti
onandcombust
ionofoxy
gen

5.Reduct
ionofoxy
gen

6.Anaer
obi
cChamber

1.Pr
oduct
ionofv
acuum

Cul
ti
vat
ioni
nvacuum wasat
tempt
edbyi
ncubat
ingcul
tur
esi
nav
acuum
dessi
cat
or.Butt
hemet
hodi
sunsat
isf
act
orybecause

-Someoxy
geni
sal
way
slef
tbehi
nd

-Li
qui
dmedi
amayboi
lov
er&sol
i
dmedi
agetdet
achedf
rom t
hepl
ate.
 2.Di
spl
acementofoxy
genwi
thot
hergases

Di
spl
acementofoxy
genwi
thot
hergasessuchasH2,He,N2orCO2
i
str
ied.Butt
hismet
hodr
arel
ypr
oducescompl
eteanaer
obi
osi
s.

A popul
arbuti
nef
fect
ivemet
hodi
stheCandl
ejar
.Int
hismet
hod
i
nocul
atedpl
atesar
epl
acedi
nsi
deal
argeai
rti
ghtcont
ainerandal
i
ght
ed
candl
ekepti
nitbef
oret
hel
i
discl
osed&seal
ed.Thebur
ningcandl
eis
expect
edt
ouseal
ltheoxy
geni
nsi
de,bef
orei
tisext
ingui
shed,butsome
oxy
genal
way
slef
tbehi
nd.Thecandl
ejarpr
ovi
desaconcent
rat
ionofCO2
whi
chst
imul
atest
hegr
owt
hofmostCapnophi
l
icbact
eri
a.

3.Absor
pti
onofOxy
genbybi
ologi
cal&chemi
calmet
hod

I
nbi
ologi
calmet
hodofoxy
genabsor
pti
on,
smal
lcl
osedsy
stem hasbeen
at
tempt
edbyi
ncubat
ional
ongwi
thaer
obi
cbact
eri
a,ger
minat
ingseedsor
choppedv
eget
abl
es.Anaer
obi
osi
spr
oducedbyt
hismet
hodi
sver
ysl
ow&
i
nef
fect
ive.

I
n chemi
calmet
hod ofoxy
gen absor
pti
on mai
nlyt
wo met
hods ar
e
f
oll
owi
ng

o Al
kal
i
nePy
rogal
l
olmet
hod

o Rosent
hal
met
hod

I
npy
rogal
l
olmet
hod,al
kal
i
nepy
rogal
l
olabsor
bsoxy
gen.I
nthi
smet
hod,
anaer
obesar
ecul
tur
edi
ntestt
ubes.Thet
estt
ubei
sthencot
tonpl
ugged
andt
heCot
tonpl
ugi
stheni
gni
ted.Af
tert
hati
tispushedi
ntot
het
estt
ube
wi
tht
hehel
pofagl
assr
od.Abov
ethatpy
rogal
l
icaci
dand1or2ml4%
sodi
um hy
droxi
dei
saddedandt
het
ubei
sseal
edwi
thr
ubberst
opper
.
Thusi
tcr
eat
esanaer
obi
ccondi
ti
onwi
thi
nthet
ube

I
nRosent
halmet
hodmi
xtur
eofchr
omi
um &H2SO4ory
ell
owphosphor
us
i
sused.

4.Di
spl
acementandcombust
ionofoxy
gen

Themostr
eli
abl
eandwi
del
yusedanaer
obi
cmet
hodi
sMcI
ntoshf
iel
d’s
j
aroranaer
obi
cjar
.Iti
sacy
li
ndr
icalv
esselmadeupofgl
assormet
alwi
th
amet
all
i
dwhi
chi
shel
dfi
rml
yinapl
acebyacl
amp.Thel
i
dhast
wot
ubes
wi
tht
aps:oneactasgasi
nletot
heractagasout
let
.Theundersur
faceof
t
hel
i
dcar
ri
esasachetcar
ryi
ngal
umi
nium pal
l
etcoat
edwi
thpal
l
adi
um.I
t
actasr
oom t
emper
atur
ecat
aly
stf
ort
heconv
ersi
onofH2&O2i
ntoH2O.

Thenew met
hodofchoi
ceofmaki
nganaer
obi
cjari
sgaspackmet
hod.
Gas pack i
s a commer
cial
l
yav
ail
abl
e di
sposabl
e env
elope cont
aini
ng
chemi
cal
swhi
chgener
ateH2&CO2ont
headdi
ti
onofH2O.

Af
tert
hepl
atesar
ekepti
njart
hegaspackenv
elopewi
thwat
eri
spl
aced
&t
hel
i
discl
osed.H2andCO2i
sli
ber
atedf
rom t
heenv
elopeandi
nthe
pr
esenceofpal
l
adi
um H2&l
eftov
eroxy
genr
eact
stof
orm wat
er.

5.Reduct
ionofOxy
genbyr
educi
ngagent

I
tisachi
evedbyusi
ngv
ari
ousr
educi
ngagenti
ncl
udi
ng1% gl
ucose,
0.
1%t
hiogl
ycol
ate,
0.1%ascor
bicaci
dand0.
05%cy
stei
ne.

Exampl
eRCM Br
oth
 6.Anaer
obi
cchamberorgl
ovebox

I
sasel
fcont
ainedanaer
obi
csy
stem wi
thpr
ovi
sionofci
rcul
ati
ngofH2,
N2&CO2wi
thi
nit&cat
aly
ticconv
ersi
onofO2t
oH2O.I
tisexpensi
ve.I
tis
r
ecommendedf
ort
otal
anaer
obi
cst
udi
es.