La f lore bacterienne dans les eaux minerales naturelles non

Dissertationsnummer:
7044
La f lore bacterienne dans les
naturelles
non
et mises
gazeif iees
denombrements de
la flore, parametres
eaux
en
minerales
boutei I les
colonies, differentiation taxonomique
physiologiques de la croissance
THESE
I
presented
l'Ecole
polytechnique
federale de Zlirich
pour l'obtention du titre de
sciences
Oocteur §s
techniques
par
Pierre
SCHWALLER
dipl.
technologie
en
alimentaire EPF-Z
n§
le 8 Janvier 1950
originaire
acceptee
du Professeur Dr. W.
du Professuer Dr.
M.
-
de Luterbach
sur
(SO)
proposition
SCHMIDT-LORENZ, rapporteur
BACHMANN,
1982
-
corrapporteur
:
de
-
\v>
-
ABSTRACT
The
current
knowledge
in several bottled waters
present
ra
cific properties is still
consequences due
to the
because this water
from natural
incomplete.
from
change
example for drinking water,
nated and bottled water
distribution nf bacterial
colony counts,
of
is
is bottled in
static
a
little known.
model
ideal
an
dynamic to
a
system, for
Natural
non-carbo¬
illustrate this phenomena,
to
large quantities
standardi¬
and in well
reproducible qualitative conditions.
zed and
CHAPTER I
I"
:
the
preliminary experiments,
parameters such
composition
as
influence of
the diluent
of
Four
french bottled mineral
waters
analytical
some
solution, shaking of the
bottle, culture media, culture methods and incubation period
mined.
C and 0)
(A, B,
were
1
charge
of each
number of
and 3 weeks after
units
colony-forming
count method and
bottling operation.
the
determined
was
using
deter¬
quantita¬
were
3 bottles
tively 8nd qualitatively investigated microbiologically, i.e.
pour
spe¬
particular, the bacteriological
In
to date
are
gene¬
and their
sources
The maximum
the surface
colony
incubation period of 21 days at 20 C, not with the
an
plate method officially prescribed in other countries. Plate Count
Agar diluted 10 fold and "Collins-Agar" produced higher colony counts
did undiluted Plate Count
than
served
one
week after
3 weeks after
ml,
bottling
CHAPTER II
rent groups could be
distinguished
the group of Gram
more
Tirst
and
isolation (both 5
was
The
heterogenous group
very
75 S lower,
in
isolated at
were
the
rough
the
four
the Pseudomonas
a
to
ranging
random
on
identification of
waters.
Five diffe¬
(20 to 50S of the
into
reaction
made
four
and
group of bacteria,
using
groups
more
the group of Gram
of
the
according
motility.
group of
30 biochemiral
than
to
unculturable
10S).
(248 strains) and the
strains)
se
:
detailled identification of
pigmented bacteria
vided
to
negative, yellow pigmented rods (6 to MS);
tive and Gram variable coccii
A
60
"Acinetobacter-Alcaligenes-Moraxella" (15 to 45S); Gram posi¬
the group
upon the
for
flora and its distribution
the bacterial
rlora)j
were
Representative colonies (1200)
!
ob¬
per ml.
three different culture media
each of
were
36,000 to 260,000 CfU per
from
these figures
18,000 to 85,000 colonies
from
Agar. The highest colony counts
bottling, ranging
negative, yellow
the Pseudomonas
and
physiological
(467
tests.
yellow pigmented rods could be subdi¬
two main
characteristics,
the oxida¬
The strains of Psoiidomon.is could be subdivided
136
-
into
four groups
duction of
according to their four
fluorescent
a
of
the first' group
was
arginine dehydro-
easily identifiable. The three
The distribution of
species.
an
glucose and alcalinisation of the medium.
were'very heterogenous and did not show
monas
the pro¬
characteristics,
main
the presence of
pigment,
lase, oxidative metabolism
Only
-
other groups
of Pseudo-
properties typical
flora
the isolated bacterial
four mineral waters showed that each water has its
own
in the
distinguishing
in all I* mi¬
"finger print",' although each group of bacteria is present
neral
waters to'a different
strains
slow
were
CHAPTER III
17
t
The
representative
The
Incubator".
or
In all
growing,-
optimal,
strains
maximal
were
and minimal
optimal temperatures varied
could grow at 37
A
and
39 C,
or
no
strains,
liquid
medium
ration
was
cells
was
found between
was
highest growth
42°C.
observed in
matter per liter).
even
in distilled water when
enough.
The strains tested
Minimal
two
a
mass
and
represen¬
diluted
A bacterial
prolife¬
the initial number of
possessed marked adaptative
(very
low nutrient concentration of medium,
directely isolated from original water cultures),
the arti¬
increase of the surface-volume ratio with glass beads could
a
tem¬
typically oligocarbotolerant.
Under certains conditions
provoke
were
possible
mixed flora
few strains
a
liquid medium. With
rates
organic
were
32 C with gene¬
the bacterial cellular
g of
low
to
days. All the strains could
(0,85
abilities and
ficial
from 20
strain could grow at
the concentration of nutrients of the
the
of
"Temperature-Gradient-
prolonged incubatory period during several weeks.
a
linear correlation
tative
water.
growth temperatures
a
of
majority
the
typical physiolo¬
a
6,5 hours. Only
to
could not be assessed after 7
grow at 0 C after
being
determined with
ration times in the range between 1,2
peratures
the waters,
this
the isolated bacteria of mineral
for
gical property
extent.
very slow
decrease of
the
generation time
of
these bacteria.
157
-
-
PCSUME
I
coiinai!!3tincnn octuellcs conccrnanl
en
<lirs (|fiircn
lmcl.t;riens-pri§3onl*f<lnn3
Innitri 1 Ira
ainsi
In
il'unc
consommation, sont
qazi5iri»5e •ekmisc
non
plwjnonione,
cc
I inns
bouteillc rcprdsente
en
et bicn
modele, id(5nl pour illustrcr
.reproductiblcs.
Aprfes avoir determine dans dcs esr.ais prdliminaires 1'influence
I
quelques paramfctres onalytiques tels
lation dr>
un
exemple
par
L'eau. min«5rale naturelle,
pen jconnu.es*
elle. ost soutirde.cn qrnndes quontiWs et dans des condi-
car
qualitatives standardises
ClIAPITRE I
iln
encore
passaqe de
ou
clos,
vase
en
on
insufrisontes.
encore
iiilierentes
syslcmc staliquc
un
a
la composition
naturclles mines
sources
Iwwttdriologiqucs
consequences
systfcme dynumiquc
de
can
de
raux
leursipropridtds speciPiques sunt
quo
pari irtilifcf,.. Inn
1'eati d'tm
In nomlirc do germcs ct
Inn
la
boutcille,
bal.oire, quatre
la solution de
que
minerales naturelles et
eoux
dilution, l'agi-
les milieux et mnlhodes de culture et
la durde incu-
qnzdifides francaises
non
(sources A, 0, C et D) ont subi des analyses microbiologiques quantitatives
qualitatives
el
1
puis
lonies
formant
et
face
crment
raison de 3 bouteilles par
a
3 semaincs
10 Tois et
"Collins-Agar"
elevi53 que Plate Count
out 6X.6 observes
36.000 et
a
avec
20 C,
livraisons,
tes nombres maximaux de
la mdthode d'ensemencement
et
non
co¬
en
sur¬
la imSthode d'ensemen-
avec
profondeur prescrite ofriciellement dans divers pays. Plate Count
Agar dilue
ves
jours
livraison et de 3
soutiraqe.
plages ont 616 obtenus
incubation de 21
une
en
aprfes leur date de
1
260.000 par
3 scmaines aprfes
le
Agar
ont produit des nombres de colonics
dilue\
non
semaine aprfes le
soutiraqe,
ml, alors qu'ils accusaicnt
soutirage,
se
plU3
Les nombres de colonies les plus Ale¬
ceux-ci
une
variant entre
diminution de 60
755
a
situant alors entre IB.000 et 85.000 colo¬
nies par ml.
C-HAPITRE II;
a
1200 colonies
representatives ont 616 isoldes statistiquement
partir de 3 milieux de culture
rentiation
quntre
(20
en
jaunc (6
45
S),
n'aynnt
deux
Cinq
eaux.
50 5J de la
a
a
HU
les coques
pu
pour
errectuer
en
grossiere de la flore bocterienne ct de
groupes distincts ont pu fttrc
flore),
S),
a
sa
une
les
:
Pseudomonas
ndgatif
et
pigmentdes
le groupe "Acinetobacter-Alcaligenes-Moraxella"
Gram
positif et Gram variable et
diffe¬
distribution dans les
formes
le groupe de3 bacttSries Gram
filre recultivdcs opre3
groupes).
premier lieu
un
(15
a
groupe de bnct£ries.
l'isolement primaire (5
a
10 S pour les
1311
ell I lerential
Uiip
tdes
hi
pnsuite entreprise
616
Lp
sioloqiques.
a
iiiux,
In
<juatre groupes
en
de
tes
les
pas
trois
distribution
danb
chaqup
que
bien que
cliaqup
typique
CuftPITRE 111
de
incubateui
a
6,5
37
j.iiotp
qend-
sui
Les soucIips de Pieudomonas
groupps ont 6t6 tres
quatre
enu
se
39 C,
mais
hdtdrogpnes
de battdries ait
v/anables.
dtd
Dans
lente
sa
ne
une
les eaux,
la
lente, propriety physiologique
Isoldes d'eaux mindrales.
rpprpsentatives ont 616 ddtermindes
a
temperatures. Les temperatures optimales
se
32
C
1,2
auec
des
temps
de
gdndration
allant
partie dps souches avait la facultd
souche n'a
prolifdrer
pu
a
42°C.
de
les
Une correlation
ne
et
deux
lineaire
rapides etaient obserues
ble
mpme
faible.
et
dans
de
1'p.iu
Les sourhet,
etaient
pnr
a
en
souchps
matieres orgamques
a
a
a
;
toutes
periode mcubatoire
une
apres
si-
sunaines.
la concentration
quide. Awec
croitre
la
temperatures
0 C
effpt
pn
de
croitre
les souches ont
pu
de
l'aide d'un
de
aucune
plusieurs
eaux,
majorlte
atteintcs apres 7 jours
a
La
"pm(jreinte diqitale",
temperatures nptimales, maximales et minimales
et
iden¬
possedaipnt
flore bactdnenne lsolee
la
propre
toutes
pt
species.
minimales n'dtaipnt pas deflnitiuement
prolongde
caiatte-
present dans les quatrp
tres
a
et
ont
In ptesenLp
glucose
de Pseudomonas
roindrales de
eaux
diotinquait par
croissance
gradient
Seule
quatre
la base de leors
facilement
souches
17
entre 20 L
heuies.
ou
Les
:
croissancp
tuaienl
hi
le premier groupe dtait
groupe
a
piqmpnteps
'jeul
bactdrips
ces
a
den bactdries
du
autrps
a
la base de deux cacaracteies
le mptabolisme oxidatif
les
souches dtait
des
piqmen-
l'argimne,
dans des proportions
mais
(I
(467 souihe.)
teat'! biochimiques et phy-
JO
propnetes habituellement typiqups
montrjil
neqalif
uiaiu
Pseuriomonas
production d'un pigment fluorescent,
la
1'alcalinlsation du milipu.
tifiable,
sur
l.i mobilitd.
reaction uxidase pt
dihydrolasp
plus de
hdlbroqpne
quatre goupes
pn
<Hre reparties
qroupp dps
chi
utilisant
on
ristiques principales,
d'une
et
l).u te>ies
des
qioupe
qioupe trPs
etre diuise
pu
pu
line iki
ion
J.iunp (24(1 southes)
etd Ptablie entre
la
masse
cellulaire bactdnen-
matiferes nutritives dans le milieu nutntif
repiesentatlues,
dins
un
litip).
distil lee
les
temps
milieu nutritlf
Une
si
dp
li-
generation les plus
liquide
dilue
(0,05
g
de
pioliTeration bactdripnnp dtiit possi¬
1 'ensemencement
testees posspdnipnt
dp
qrandes
typiquement oliqotaibotoldrantes.
initial
facultes
dtait
assez
d'adaptat
ion
-
'mils
ceilainos conditions
tives,
I lorn
medium;
sor
voliimc rriVprovoifmr
"mixlr"
mi
par
unc
137
-
(Iron faiblrs cnneonlialinnn
isolec directoiiionl
milieu do
laborntoiro),
1'adjonction dc billes
diminution du
temps
de
111
maliiMos nutri¬
d'oau minerale
sain
I'auc/mcnlal
do
do
verro
feneration
ion
dnns
de
CC3
passaoo
inter¬
rapport nurTacc-
la bouteille pouvait
bacteries.