FoodMicrobiology, 1997, 14, 143151

ORIGINALARTICLE
Ariskassessment studyof Bacillus
cereuspresent inpasteurizedmilk
S. Notermans
1
*, J . Dufrenne
1
, P. Teunis
1
, R. Beumer
2
,
M. teGiffel
2
andP. PeetersWeem
3
Due to the heat resistance of Bacillus cereus, its potential pathogenic character, the
capability to grow in milk and reported diseases upon consumption of dairy products, the
organism should be considered as hazardous in pasteurized milk. Human exposure
experiments based on (1) enquiring about the storage conditions (time and temperature) of
pasteurized milk in households in The Netherlands and (2) performing of storage tests at 6,
8, 10 and 12C, respectively, demonstrated the following: pasteurized milk is consumed
within 212 days after pasteurization and is stored at temperatures varying from <513C.
Visually spoiled milk is not consumed anymore. Based on the distribution of both the
storage time and temperature of milk in private households, approximately 7 and 4%of the
total portions of milk consumed in The Netherlands could contain >10
5
and >10
6
B. cereus
ml
1
, respectively. Taking into account the high portion of milk consumed (approximately
10
9
10
10
portions/year) and the absence of epidemiological evidence that milk is causing
foodborne diseases due to B. cereus, the doseresponse relationship of this organism may
need to be considered. Results of the storage tests, based on the worst case situation,
provided useful information on the effect of the risk factors, storage temperature and time,
to human exposure. However, such information can be obtained more easily by applying
predictive models. Models also enable us to find out the initial number of spores which can
be considered as a third riskfactor. 1997Academic Press Limited
Received:
15J uly 1996
1
National Institute
of Public Health
Introduction produced, and that safety has economi cal
and the
consequences. Therefore, there i s al so a Environment, P.O.
Box 1, 3720BA
Over the l ast 20 years, there has been growi ng i nterest i n the appl i cati on of ri sk
Bilthoven, The
i ncreased acti vi ty i n the devel opment of con- assessment, or at l east el ements of i t, i n the
Netherlands
cepts for producti on of safe food to protect the context of GMP/HACCP.
2
Agricultural
University of
consumer. Now, i t becomes i ncreasi ngl y cl ear Ri sk assessment i s a quanti tati ve esti -
Wageningen, P.O.
that the appl i cati on of good manufacturi ng mati on of the probabi l i ty of occurrence of a
Box 8129, 6700EV
practi ces (GMP), fol l owed by process control , (foodborne) hazard. I nformati on about the Wageningen, The
Netherlands
based on the hazard anal ysi s cri ti cal control probabi l i ty of occurrence of a hazard i s
3
Regional
poi nt (HACCP) concept are prevai l i ng. How- becomi ng i ncreasi ngl y i mportant for mana-
Inspectorate for
ever, appl i cati on of GMP/HACCP makes i t geri al deci si ons. However, i t shoul d be noted
Health Protection,
Oostergoweg 2,
expl i ci t that absol utel y safe food can not be that at thi s moment l i ttl e experi ence exi sts i n
8932PG
the use of ri sk anal ysi s i n the producti on of
Leeuwarden, The
safe food. So far, there have been onl y a few
Netherlands
*Correspondi ng author.
0740-0020/97/020143+09 $25.00/0/fd960076 1997 Academi c Press Li mi ted
144 S. Notermans et al.
formal , food-rel ated, mi crobi ol ogi cal ri sk- Ri sk assessment i s the esti mati on of the
probabi l i ty that an adverse effect wi l l occur assessment studi es. One of these studi es
deal t wi th ri sk assessment of the use of to the consumer (i n thi s speci al case caused
by B. cereus, present i n pasteuri zed mi l k) and cracked eggs (Todd 1996) and another study
deal t wi th a heal th-ri sk assessment of Lis- i s a component of ri sk anal ysi s. The process
of ri sk assessment consi sts of the fol l owi ng teria monocytogenes (Farber et al . 1996).
These studi es demonstrate that ri sk-assess- steps: (1) hazard i denti fi cati on, (2) exposure
assessment, (3) doseresponse assessment ment resul ts can easi l y be made subservi ent
to HACCP. Thi s especi al l y appl i es to the so- and (4) ri sk characteri zati on.
Hazard i denti fi cati on i s the i denti fi cati on cal l ed cri ti cal control poi nts (CCPs) whi ch
have to be i denti fi ed i n the HACCP approach. as to whether B. cereus may have the poten-
ti al to cause an adverse heal th effect upon At these CCPs, cri ti cal l i mi ts shoul d be speci -
fi ed i n order to meet decl ared end-product consumpti on of pasteuri zed mi l k. For i denti -
fi cati on of B. cereus as a hazardous organi sm speci fi cati ons (Notermans and Mead 1996).
Or i n other words, CCPs cri ti cal l i mi ts have i n pasteuri zed mi l k, the fl ow sheet as pro-
posed by Notermans and Mead (1994) has to be set whi ch guarantee an acceptabl e l ow
ri sk that the fi nal product may cause harm. been appl i ed.
Exposure assessment i s the quanti tati ve For a (processed) peri shabl e product, such as
pasteuri zed mi l k, cri ti cal l i mi ts may be, for esti mati on of the dose of B. cereus to whi ch
the consumers are exposed at ti me of con- exampl e, the storage ti me and temperature.
Ri sk assessment can provi de thi s sumpti on. To gai n i nformati on on human
exposure, storage tests and studi es on the i nformati on.
Regul atory agenci es are i nvol ved i n the storage condi ti ons of pasteuri zed mi l k i n the
home have been appl i ed. demonstrati on that the producers are adher-
i ng to exi sti ng regul ati ons and di recti ves. Doseresponse assessment i s the trans-
l ati on of exposure ti me i nto di sease. I t i s the They are al so responsi bl e for setti ng end-
product speci fi cati ons, whi ch are the basi s to process of obtai ni ng quanti tati ve i nformati on
on the probabi l i ty of human i l l ness fol l owi ng whi ch producers set thei r cri ti cal l i mi ts at
CCPs. Now, wi th the i ntroducti on of the exposure to B. cereus. Doseresponse
rel ati onshi ps may be gai ned from epi demi ol - GMP/HACCP concept i n food processi ng,
i nspecti on of the control system and gui dance ogi cal anal yses of foodborne di seases caused
by B. cereus. may become an addi ti onal task for the auth-
ori ti es. Gui dance compri ses, among others, Ri sk characteri zati on i s the process of
i denti fi cati on and quanti fi cati on of the fac- taki ng the i ni ti ati ve i n the process of com-
muni cati on between producers and con- tors i nvol ved i n the ri sk of bei ng exposed to
unacceptabl e l evel s of B. cereus at ti me of sumers. A cl ear exampl e i s the consumer
behavi our and the possi bl e consequences for consumpti on of the mi l k.
the safety of a food product i n terms of setti ng
cri ti cal l i mi ts at CCPs. Regul atory agenci es
Enquiries carried out in households
may al so benefi t from ri sk assessment.
I n thi s paper, a ri sk-assessment study of Enqui ri es were carri ed out to obtai n i nfor-
mati on regardi ng the storage condi ti ons Bacillus cereus present i n pasteuri zed mi l k i s
presented. (ti me and temperature) of pasteuri zed mi l k
i n househol ds. Two hundred and seventy-
three pri vate househol ds were vi si ted, and
the storage ti mes of opened jars of mi l k were
MaterialsandMethods
determi ned. I n addi ti on, parti ci pants were
asked whether vi sual l y-spoi l ed mi l k was con-
Definitions for riskassessment
sumed or not. The average temperature of
refri gerators was measured i n 1993 by the Defi ni ti ons used are based on the defi ni ti ons
recentl y presented by Notermans et al . Regi onal I nspectorate for Heal th Protecti on
i n Leeuwarden (de Lezenne Coul ander 1994). (1996).
A risk assessment study of Bacillus cereus present in pasteurized milk 145
They tested 125 refri gerators and thei r Numbers. From day 2, the mi l k was tested
for the number of B. cereus by prepari ng 10- resul ts have been used i n thi s study.
fol d di l uti on of the mi l k i n peptone physi o-
l ogi cal sal i ne (PPS) and pl ati ng out these
Storage tests
di l uti ons on MEYP fol l owed by i ncubati on at
30C for 182 h. Typi cal col oni es were tested Every 2 weeks, from March 1995 unti l
February 1996, two packages of freshl y-pas- as for the MPN, by testi ng three col oni es for
each sampl e di l uti on that presented coun- teuri zed mi l k (fat content 3%) were obtai ned
from si x di fferent mi l k-processi ng pl ants i n tabl e col oni es.
The Netherl ands. The mi l k was produced on
Monday and del i vered at the l aboratory on
Enumeration of the total count present
Tuesday havi ng been stored at 6C. I mmedi -
in milk
atel y after del i very (day 1), two packages of
one processi ng pl ant were chosen and mi xed.
To determi ne the total bacteri al count pre-
The numbers of B. cereus present were esti -
sent i n stored mi l k sampl es, 10-fol d di l uti ons
mated by usi ng a most probabl e number
of mi l k i n PPS were pl ated on pl ate count
(MPN) procedure. The rest of the mi l k was
agar (Di fco). The pl ates were i ncubated at
di vi ded i nto four porti ons of 200 ml , put i nto
30C for a peri od of 3 days.
300 ml steri l e Erl enmeyer fl asks and pl aced
i n thermostati cal l y-control l ed water-baths at
temperatures of 6, 8, 10 and 12C, respect-
Results
i vel y. At di fferent i nterval s, dependi ng on the
storage temperature, the numbers of B.
Hazard identification
cereus present were counted usi ng a pl ati ng
techni que, unti l vi sual spoi l age (coagul ati on) B. cereus i s a wel l -establ i shed, foodborne
pathogen. Anal ysi s of reported foodborne was observed. The maxi mal storage ti me was
24 days. I n total , 38 mi xed sampl es, si x to di seases reveal s that the organi sm i s fre-
quentl y di agnosed as the cause of gastroi n- seven sampl es from each pl ant, were used i n
the storage tests. testi nal di sorders (Kramer and Gi l bert 1989).
The organi sm i s ubi qui tous i n nature, and as
a consequence, present i n ani mal feed and
Enumeration of B. cereus present in milk
fodder (Kramer and Gi l bert 1989). Due to i ts
rapi d sporul ati ng capaci ty, the organi sm eas- MPN. From every mi l k sampl e at day 1,
porti ons of 3100 ml , 310 ml and 31 ml i l y survi ves i n the envi ronment and duri ng
i ntesti nal passage i n cows. The organi sm con- were taken and i ncubated at 30C. After
i ncubati on for 182 h, the porti ons were tami nates raw mi l k vi a faeces and soi l . I t has
been demonstrated that addi ti onal sources of pl ated on manni tol egg yol kpol ymyxi n agar
(MEYP, Mossel et al . 1967) and i ncubated at mi l k contami nati on are the pi pel i nes and
mi l k tanks at the processi ng pl ants (Van 30C for 18 2 h. From each pl ate wi th typi -
cal col oni es resembl i ng B. cereus, three Heddeghem and Vl aemynck 1993; Te Gi ffel
et al . 1996). col oni es were pi cked and streaked on to tryp-
tone soya agar (TSA) pl ates and then i ncu- Due to thei r heat-stabl e spores, B. cereus
easi l y survi ves the pasteuri zati on process. bated at 30C for 182 h. From the TSA
pl ates, brai n hearti nfusi on (BHI ) broth Resul ts obtai ned i n thi s study reveal ed that
day-ol d pasteuri zed mi l k contai ned about 10 tubes were i nocul ated, and i ncubated at 30C
for 242 h. Four confi rmati on tests were per- B. cereus organi sms per 100 ml (Tabl e 1) and
that al l 38 sampl es of each two 1 l packages formed on the B. cereus i n the BHI tubes:
VogesProskauer (+); gel ati ne hydrol ysi s (+); contai ned B. cereus. The number of spores
present seems to be seasonal l y dependent gl ucose fermentati on (+) and ni trate
reducti on (+). The numbers of B. cereus were wi th the hi ghest contami nati on occurri ng
duri ng the grazi ng peri od (Meer et al . 1991, then esti mated by usi ng the MPN method of
De Man (1974). Te Gi ffel et al . 1995). From l i terature anal y-
146 S. Notermans et al.
si s of consumer i l l nesses, i t i s evi dent that temperature, (Chri sti ansson et al . 1989,
Hel my et al . 1984, Cri el l y et a. 1994) and dai ry products account for a substanti al pro-
porti on of cases of foodborne di seases. some of these reports i ndi cate the producti on
of enterotoxi n i n mi l k by B. cereus. Recent Several outbreaks have been reported i n
whi ch mi l k or mi l k-rel ated products contai n- studi es reveal ed that the organi sm wi l l grow
at temperatures as l ow as 5C (Meer et al . i ng B. cereus were suggested to be the cause
of the di sease (Bannerjee and Bl ack 1986, 1991). As a consequence of al l the above men-
ti oned aspects, the presence of B. cereus i n Bryan 1983, Bul yba et al . 1973, WHO Sur-
vei l l ance Programme Report 1995). Anal ysi s mi l k shoul d be consi dered as hazardous.
of B. cereus strai ns i sol ated from mi l k
reveal ed that al most al l strai ns tested pro-
Exposure assessment
duced enterotoxi n (Granum et al . 1993,
Ogi hara et al . 1992, van Netten et al . 1990).
To assess human exposure to B. cereus pre-
I t has al so been cl ai med that the toxi n i s pro-
sent i n pasteuri zed mi l k, storage tests were
duced at al l storage temperatures of mi l k
carri ed out. Addi ti onal l y enqui ri es were
al l owi ng growth of B. cereus (Sutherl and
made to l earn the storage condi ti ons of paste-
1993).
uri zed mi l k i n domesti c househol ds.
To cause di sease, B. cereus has to prol i fer-
ate i n pasteuri zed mi l k. B. cereus i s an auto-
trophi c organi sm, and does not requi re a ri ch Enquiries made in households. The enqui r-
i es reveal ed that the storage ti me of pasteur- substrate for mul ti pl i cati on. Several reports
have shown that the organi sm can prol i ferate i zed mi l k vari ed from 212 days. The fre-
quency di stri buti on of the storage ti me i s rapi dl y i n mi l k, dependi ng on the storage
Table1. I ni ti al numbers of B. cereus present i n mi l k 1 day after pasteuri zati on as determi ned usi ng
a most probabl e number method appl yi ng three 10-fol d di l uti ons i n tri pl i cate
Peri od of sampl i ng Number of sampl es B. cereus count (l og
10
cfu 100 ml
1
)
MarchJune, 1995 12 0460046
a
Jul yOctober, 1995 16 140047
NovemberFebruary, 1996 12 093029
Total 38 096057
a
Standard devi ati on.
Table2. Storage ti me of pasteuri zed mi l k i n The Netherl ands
Storage ti me (days after Number of sampl es P
t
(Probabi l i ty that mi l k i s stored
pasteuri zati on of the mi l k) for the i ndi cated ti me)
1 0 0
2 6 0022
3 16 0059
4 43 0158
5 43 0158
6 33 0121
7 39 0143
8 42 0154
9 15 0055
10 12 0044
11 14 0051
12 10 0037
13 0 0
Total 273 1000
A risk assessment study of Bacillus cereus present in pasteurized milk 147
8
Days
L
o
g
1
0

B
.
c
e
r
e
u
s
16
2
6
5
4
3
1
2 4 8 12 20 0
(c)
18 6 10 14 22 24
7
Days
L
o
g
1
0

B
.
c
e
r
e
u
s
16
2
6
5
4
3
1
2 4 8 12 20 0
(a)
18 6 10 14 22 24
8
Days
L
o
g
1
0

B
.
c
e
r
e
u
s
16
2
6
5
4
3
1
2 4 8 12 20 0
(d)
18 6 10 14 22 24
8
Days
L
o
g
1
0

B
.
c
e
r
e
u
s
16
2
6
5
4
3
1
2 4 8 12 20 0
(b)
18 6 10 14 22 24
7
7 7
Figure 1. Resul ts of storage tests carri ed out wi th pasteuri zed mi l k. Porti ons of pasteuri zed mi l k
were stored at 6 (Fi g. 1(a)) 8 (Fi g. 1(b)), 10 (Fi g. 1(c)) and 12C (Fi g. 1(d)), respecti vel y. From day 2, the
i ncubated mi l k sampl es were tested for the numbers of B. cereus ml
1
unti l obvi ous spoi l age occurred.
The arrows i ndi cate the fi rst sampl es showi ng spoi l age. Spoi l ed sampl es were not tested further and
have been excl uded. The shaded areas i n the fi gures i ndi cate the consumpti on peri od and the sampl es
contai ni ng >10
5
B. cereus ml
1
.
presented i n Tabl e 2. I t can be seen that the Storage tests. Resul ts of the storage tests
are presented i n Fi g. 1(a)(d). At each of the frequency di stri buti on does not show a nor-
mal di stri buti on. A cl ear shoul der i s seen at i ncubati on temperatures the number of B.
cereus i ncreased duri ng the storage peri od. the end of the storage peri od.
Tabl e 3 shows the frequency di stri buti on Maxi mum numbers, exceedi ng 10
7
cfu ml
1
,
were observed before overt spoi l age was of the temperatures of refri gerators i n Dutch
pri vate househol ds. observed. The shaded areas i n the fi gures
Table 3. Average storage temperatures of pasteuri zed mi l k i n The Netherl ands (de Lezenne
Coul ander, 1994)
Temperature (C) of refri gerators Numbers of refri gerators wi thi n P
T
(probabi l i ty that the mi l k i s
used to store mi l k the i ndi cated temperature range stored at the i ndi cated
temperature range)
<5 37 0296
5<7 52 0416
7<9 32 0256
9<11 2 0016
11<13 2 0016
Total 125 1000
148 S. Notermans et al.
i ndi cated the peri od of consumpti on where 0341, Tabl e 2). The resul ts of these cal cu-
l ati ons are presented i n Tabl e 4. I t reveal s numbers of B. cereus exceeded 10
5
cfu ml
1
.
Spoi l age of mi l k was not observed at a that at ti me of consumpti on approxi matel y
7% of the porti ons of pasteuri zed mi l k con- storage temperature of 6C. At 8, 10 and 12C
the fi rst sampl es showi ng spoi l age were sumed contai ned >10
5
cfu B. cereus ml
1
.
The same cal cul ati ons were made for other observed after 12, 8 and 6 days, respecti vel y.
Because spoi l ed mi l k was not consumed, l evel s of contami nati on and are presented i n
Tabl e 5. these sampl es were excl uded from the
resul ts. I n The Netherl ands there are about 1510
6
consumers of pasteuri zed mi l k. I f each of I t was observed that at i ncubati on tem-
peratures of 6, 8 and 10C, the total bacteri al them dri nk 50500 porti ons of 100 ml of mi l k
per year, about 10
9
10
10
porti ons of 100 ml of count was si mi l ar to the B. cereus count
(resul ts not presented), suggesti ng that B. mi l k are consumed. The cal cul ati ons suggest
that 7 and 4% of these porti ons contai n >10
5
cereus was the domi nant bacteri al fl ora pre-
sent. At the i ncubati on temperature of 12C, and >10
6
B. cereus ml
1
, respecti vel y.
the total count was much hi gher than the
number of B. cereus present (resul ts not
presented). Doseresponse assessment
Kramer and Gi l bert (1989) summari zed a
l arge number of foodborne outbreaks caused Exposure calculations. From the resul ts
presented i n Tabl es 2 and 3 and i n Fi g. by B. cereus. The resul ts showed that the
l evel s of B. cereus present i n foods causi ng a 1(a)(d), the probabi l i ty was cal cul ated that
mi l k at ti me of consumpti on contai ned >10
5
di arrhoeal syndrome vari ed from 12
10
3
10
8
organi sms g
1
; the medi an val ues was cfu ml
1
. For thi s cal cul ati on, the resul ts of
the storage tests between 2 and 13 days of around 10
7
organi sms. Level s of B. cereus
present i n food causi ng an emeti c syndrome storage (consumpti on peri od) were the
starti ng poi nt. From thi s part, the fracti on vari ed from <10
4
>10
10
g
1
wi th a medi um
val ue of 1010
7
g
1
. was taken contai ni ng sampl es wi th a count
of >10
5
ml
1
B. cereus (i n Fi g. 1(b) i t i s the I nfecti on experi ments have al so been
carri ed out wi th ani mal s and human vol un- fracti on between 7 and 13 days of storage and
amounts of 0634). These fracti ons were mul - teers. Cats and dogs fed experi mental di ets
contai ni ng more than 10
5
B. cereus g
1
devel - ti pl i ed wi th the probabi l i ty that mi l k i s
stored between 57, 79, 911 and 1113C, oped severe di arrhoea and dehydrati on
(Ni kode

musz 1967). The human vol unteer respecti vel y (for storage between 79C thi s
fracti on i s 0256, Tabl e 3) and mul ti pl i ed study carri ed out by Hague (1995) supported
the noti on that B. cereus al so i nduced di ar- wi th the probabi l i ty that the mi l k i s stored
between the i ndi cated days (for a storage rhoea i n humans. However, Dack et al .
(1954), fai l ed to produce food-poi soni ng symp- peri od between 7 and 13 days thi s fracti on i s
Table 4. Cal cul ati on of the probabi l i ty that pasteuri zed mi l k contai ns >10
5
B. cereus ml
1
at ti me of
consumpti on
Storage temp. (C) 6 8 10 12
(and range) (5<7) (7<9) (9<11) (11<12)
Fracti on
a
>10
5
ml
1
0 0634 0755 0431
P
T
b
0416 0256 0016 0016
P
t
c
0 0341 0762 0847
P>10
5
= (Fracti onP
T
P
t
) 0 0055 0009 0006=007
a
Fracti on of the mi l k contai ni ng >10
5
B. cereus ml
1
i n the peri od of consumpti on (see Fi g. 1(a)1(d)).
b
Fracti on of the mi l k stored i n the rel ated temperature range (see Tabl e 3).
c
Fracti on of the mi l k stored i n the rel ated peri od (see Tabl e 2).
A risk assessment study of Bacillus cereus present in pasteurized milk 149
toms wi th B. cereus. Recentl y, Langevel d et there are suggesti ons that B. cereus may not
al ways cause harm (Dack 1954, Langevel d et al . (1996) carri ed out a quanti tati ve human
vol unteer study. Duri ng a peri od of 3 weeks al . 1996), i t has been consi dered that a
number of 10
5
(toxi geni c) B. cereus present i n the parti ci pants were exposed to B. cereus by
dri nki ng porti ons of pasteuri zed mi l k whi ch pasteuri zed mi l k shoul d be regarded as
hazardous to the consumer. Our resul ts dem- had been stored for 314 days at 75C. The
total numbers of B. cereus i ngested vari ed i n onstrate that 7% of the porti ons of mi l k at
ti me of consumpti on coul d show a B. cereus that study from 10
5
>10
8
organi sms (10
3
>
10
6
ml
1
). No i ndi cati ons were obtai ned that count exceedi ng thi s val ue.
these organi sms caused harm to the vol un-
teers. The di screpancy between reported and
Riskcharacterization
anal ysed outbreaks, and certai n human vol -
unteer studi es may be expl ai ned by assumi ng The mai n factors i nvol ved i n the ri sk of bei ng
that not al l strai ns of B. cereus have the exposed to unacceptabl e l evel s of B. cereus at
potency to cause food-poi soni ng. ti me of consumpti on of mi l k compri se the
I n regul ati ons of most European countri es, i ni ti al count of spores present, the storage
numbers of 10
4
10
5
cfu B. cereus ml
1
of a temperature and the storage ti me. I n thi s
product at expi ry date, are set as cri ti cal study, onl y i nformati on about the effect of
l i mi ts for acceptance of a parti cul ar food. Al so storage ti me and storage temperature were
for di agnosi ng foodborne di sorders resem- assessed. Tabl e 6 presents the effect of both
bl i ng those caused by B. cereus, numbers of factors on a fi nal l evel of 10
5
cfu B. cereus ml
1
B. cereus >10
5
are regarded as hi ghl y of mi l k. The cal cul ati ons are based on the
i ndi cati ve. worst-case condi ti ons. I n The Netherl ands,
From a hygi eni c poi nt of vi ew, the pres- pasteuri zed mi l k has to be stored at 7C for
ence of B. cereus i n food i n excess of 10
5
/10
6
maxi mal 7 days. Under these condi ti ons the
organi sms g
1
i s consi dered not consi stent number of B. cereus present at expi ry date
wi th the general l y-accepted pri nci pl es of good wi l l not be >10
5
cfu ml
1
. Tabl e 6 al so shows
food practi ce. predi cted resul ts based on the mathemati cal
I n concl usi on, B. cereus i s a wel l -recog- model appl i ed by Zwi eteri ng et al . (1996).
ni zed pathogeni c organi sm. Al though, no Compari ng the resul ts reveal s that both
cl ear doseresponse data are avai l abl e, and approaches provi de fai rl y equal resul ts. I t
has to be noted that i n the model of
Zwi eteri ng et al . (1996) the l ag phase was not
Table 5. Exposure to B. cereus after
taken i nto account.
consumpti on of pasteuri zed mi l k (based on the
cal cul ati ons as demonstrated i n Tabl e 4)
Exposure dose (cfu ml
1
) Probabi l i ty
Discussion
>10 99%
>10
2
21%
I n thi s paper the formal process of ri sk
>10
3
14%
>10
4
11% assessment has been worked out i n detai l for
>10
5
7%
B. cereus and pasteuri zed mi l k. Thi s exampl e
>10
6
4%
was chosen because most aspects of ri sk
>10
7
<1%
assessment are represented.
Table6. Ti me i n days for B. cereus to mul ti pl y to 10
5
cfu ml
1
at di fferent temperatures usi ng storage
tests and predi cti ve mi crobi ol ogy, based on the most worse case si tuati on
Method 6C 8C 10C 12C
Storage tests
a
138 68 42 26
Predi cti ve mi crobi ol ogy
b
129 72 46 32
a
I ni ti al count of B. cereus 009 ml
1
(see Tabl e 1).
b
Based on the model of Zwi eteri ng et al . (1996); i ni ti al
count of B. cereus 01 ml
1
.
150 S. Notermans et al.
The hazard-i denti fi cati on study cl earl y for the present i t remai ns advi sabl e to adhere
to exi sti ng regul ati ons on maxi mal numbers reveal ed that B. cereus present i n mi l k may
cause harm to the consumer. As a conse- of B. cereus present i n ready-to-eat food.
Ri sk characteri zati on provi des i nformati on quence, assessment of human exposure to B.
cereus vi a consumpti on of mi l k was justi fi ed. about the factors i nvol ved i n unacceptabl e
exposure to B. cereus vi a consumpti on of pas- Exposure anal ysi s was carri ed out based
on househol d enqui ri es regardi ng storage teuri zed mi l k. Factors that cl earl y determi ne
the ri sk of bei ng exposed to an excessi ve condi ti ons of pasteuri zed mi l k and on storage
tests of mi l k at several i ncubati on tempera- number of B. cereus are the i ni ti al count after
pasteuri zati on, the storage ti me and the stor- tures. I n the cal cul ati ons, storage ti mes and
storage temperatures were consi dered to be age temperature. The present study onl y pro-
vi des i nformati on about the effect of storage i ndependent, whi ch i s obvi ousl y not true. I t
may be assumed that mi l k stored at a hi gh ti me and the storage temperature. However,
such i nformati on can al so be obtai ned by temperature wi l l be stored for a shorter
peri od si mpl y due to spoi l age. However, vi su- appl yi ng predi cti ve methodol ogy, as has been
demonstrated recentl y by Zwi eteri ng et al . al l y-spoi l ed mi l k i s not consumed and was
excl uded from the cal cul ati ons. (1996). Compari ng the resul ts obtai ned
appl yi ng the model used by Zwi eteri ng et al . For i nterpretati on of the human exposure,
i t i s necessary to have quanti tati ve i nfor- (1996), and the resul ts found i n thi s study
based on storage testi ng reveal s that defi ni - mati on about the consequences of bei ng
exposed. I n thi s study, the necessary i nfor- ti ve i nformati on i s as yet not attai nabl e. The
mai n reason i s that the model of Zwi eteri ng mati on was obtai ned from a l i terature survey
and col l ecti on of addi ti onal i nformati on. et al . (1996) i s based on the most worse-case
si tuati on. However, the model al l ows at l east Resul ts obtai ned reveal ed that al though B.
cereus can cause foodborne i l l ness, no cl ear cal cul ati ons of the rel ati ve contri buti ons of
several ri sk factors, i ncl udi ng the effect of the resul ts are avai l abl e to esti mate a dose
response rel ati onshi p. Addi ti onal l y, there i ni ti al count of B. cereus.
Consi deri ng that consumpti on of pasteur- exi st evi dence that not al l B. cereus wi l l cause
foodborne i l l ness (Dack et al . 1954; i zed mi l k contai ni ng >10
5
cfu of B. cereus ml
1
may cause harm to the consumer suggests Langevel d et al . 1996). Based on anal ysi s of
foodborne i l l ness caused by B. cereus i n the that the expi ry date of pasteuri zed mi l k pro-
duced i n The Netherl ands can be set at 7 past, safety cri teri a have been set; these vary
from 10
4
10
5
g
1
of a food product. Our cal cu- days based on a storage temperature of 7C.
Al though these condi ti ons have been made l ati ons demonstrate that 11% of the porti ons
of mi l k consumed i n The Netherl ands mandatory i n the past a di scussi on between
producers of pasteuri zed mi l k and consumer contai ns >10
4
B. cereus ml
1
, whi l e 7% of the
porti ons contai n >10
5
ml
1
. The resul ts organi zati ons shoul d be i ni ti ated how to
i mprove the possi bl e undesi red si tuati on of obtai ned cl ari fi ed that a substanti al porti on
of consumed pasteuri zed mi l k does not meet bei ng exposed to an excessi ve number of B.
cereus, as i s demonstrated i n thi s research. the l egal cri teri a at ti me of consumpti on. I t
has been demonstrated that the storage con-
di ti ons are an i mportant factor, and that
Dutch consumers do not al ways meet the pre-
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