Home Delivery Project Rep

Microbiological Diagnostic Unit
Public Health Laboratory

The University of Melbourne
November 2003 Page 1 oI 53

Home DeIivery Project Report

Project management

Dr GeoII Hogg, Director, MDU PHL
Agnes Tan, Senior Scientist, MDU PHL
Mr Brian Cusack, Food SaIety Unit, Department oI Human Services, Victoria

MDU PHL Project Team

Sheila Beaton
May Chai
Gina Holland (Project OIIicer)
Athanasia Petalotis
Nela Subasinghe
Agnes Tan (Project Co-ordinator)

Logistic support provided by
Media Preparation Unit, University oI Melbourne (Supply oI culture media)
Michael Carmody, MDU PHL (IT)
MDU PHL
HOME DELIVERY PROJECT REPORT University oI Melbourne

TabIe of Contents

Table oI Contents .................................................................................................................................... 2
Acknowledgments................................................................................................................................... 3
Executive summary................................................................................................................................. 4
Supermarket study.......................................................................................................................... 4
Challenge tests................................................................................................................................ 5
Challenge test vs Predictive models ............................................................................................... 5
Application oI Predictive models oI microbial growth to delivery data......................................... 5
Conclusions and Recommendations............................................................................................... 6
Introduction............................................................................................................................................. 7
Collaborators....................................................................................................................................... 7
Objectives ........................................................................................................................................... 8
Project outcomes................................................................................................................................. 8
Study Plan........................................................................................................................................... 8
Stage 1: ........................................................................................................................................... 8
Stage 2: ........................................................................................................................................... 8
Stage 3: ........................................................................................................................................... 8
Stage 4: ........................................................................................................................................... 8
1. Establishing partnerships and method development ........................................................................... 9
1.1 Estabishing partnerships ............................................................................................................... 9
1.2 Method development .................................................................................................................... 9
2. Supermarket study - Temperature proIiles oI home delivered Ioods ................................................ 11
2.1 Study period................................................................................................................................ 11
2.2 Sample collection........................................................................................................................ 11
2.3 Collection oI temperature proIiles .............................................................................................. 11
2.4 Home delivery arrangement........................................................................................................ 12
2.5 Findings ...................................................................................................................................... 13
2.5.1 Delivery times ..................................................................................................................... 13
2.5.2 Temperature proIiles ........................................................................................................... 14
2.5.3 Impact oI Iood holding aIter shelI selection on temperature proIiles.................................. 17
3. Challenge Tests ................................................................................................................................. 21
3.1 Results oI Challenge tests ........................................................................................................... 22
4. Challenge tests results vs Predictive models ..................................................................................... 24
4.1 Methods oI Comparison.............................................................................................................. 24
4.1.1 Qualitative comparison........................................................................................................ 24
4.2 Comparison oI Exponential Growth Rates. ................................................................................ 26
4.3 Application oI Predictive models oI microbial growth to delivery data..................................... 27
4.3.1 Predicting growth in home delivered Ioods......................................................................... 27
5. Conclusions ....................................................................................................................................... 29
ReIerences ............................................................................................................................................. 31

MDU PHL

AcknowIedgments
The project was Iunded by the Victorian Department oI Human Services. It was conducted by the
Microbiological Diagnostic Unit Public Health Laboratory in collaboration with the two major supermarket
chains (Coles and Woolworths/SaIeway), local government environmental health oIIicers in Victoria,
Queensland and the Northern Territory and the University oI Tasmania.

The assistance provided by the Iollowing participants was crucial to the success oI this project.
Department oI Human
Services
Mr Brian Cusack, Food SaIety Victoria
Mr Gary Smith, Food SaIety Victoria
Local Government (Victoria) Mr Ross Williams, Frankston City Council
Ms Andrea Lombard, Frankston City Council
Coles Supermarkets Mr Richard Uglow
Mr Jon Davidson
Store and deli managers (Carlton, Forest Hill, Frankston, Casuarina,
Cairns)
Woolworth Supermarkets

Mr Michael Thompson
Mr Alan Fagerland
Mr Damier Bakin
Mr Louie Tseukatos
Mr Dean Gutske
Store and deli managers (Box Hill, Bulleen, The Pines, Casuarina,
Cairns)
Household contacts Mrs Mari Sist
Ms Sandra Sist
Mrs Suzie McBride
Mrs Maria Toumbourou
Mrs Margaret Walton
Queensland Mr Kerry Bell, Queensland Health
Mr Stuart Heggie, Cairns Health Department
Northern Territory Ms Tracy Ward, Casuarina Health Department
Ms Sara Robinson
University oI Tasmania Dr Tom Ross, Senior Lecturer, School oI Agricultural Science

We thank Coles and Woolworths Supermarkets Ior the donation oI the Ioods 'purchased and Ior meeting
the cost oI home delivery.

MDU PHL

Executive summary
This project set out to provide a descriptive assessment oI the risk posed by un-reIrigerated delivery oI
perishable Ioods. The Iindings will assist health authorities in the Iormulation oI a Iood saIety plan Ior the
delivery oI chilled Ioods to private homes and may contribute to the development oI uniIorm transport
requirements across all jurisdictions. It was not intended to be a detailed statistical assessment.
Supermarket study
The study was carried out using nine (9) stores six in Victoria, two in Queensland and one in the Northern
Territory. Eight Ioods (cheese, Iish, Iruit, milk, mince, pie, deli meat, peas) were purchased Irom these
stores over 5 consecutive days. Their temperatures were monitored, Irom the time oI selection Irom the
shelves to the time oI receipt in the home.
We Iound that
Deliveries Irom Melbourne stores were diIIerent Irom interstate stores in that
The median delivery times were shorter, but
The temperature increases observed in Ioods were higher.
In Victoria, delivery times can be as long as 5 hours but the time spent during road transport was
only about 30 minutes. The rest oI the time was spent on trolleys in the supermarket, awaiting
delivery.
There was no correlation between the maximum delivery times and the number oI weekly deliveries
made. Delivery times were more likely to be aIIected by
Individual store practice and
The time oI purchase Ioods bought in the early morning tended to have longer delivery
times.
In most stores, Ioods were not reIrigerated aIter the check-out. For these Ioods,
The longer the delivery, the greater the temperature increase observed.
The rise in temperature was greater when the outside air temperature was higher.
A combination oI hot days and long delivery times can result in very large temperature
increases.
Some stores had a system oI reIrigerating Ioods aIter purchase. The beneIit oI reIrigeration was only
evident approximately 40 minutes to 1 hour into the delivery time when a reduction in most Iood
temperatures was recorded. Much lower temperature rises were observed in Ioods purchased Irom
these stores.
Very good temperature control was noted with Ioods placed in insulated 'Cold Packs Irom the time
oI selection Irom the shelI to delivery to the householder.
MDU PHL

ChaIIenge tests
Duplicate sets oI 5 Ioods were challenged with three strains each oI Bacillus cereus, Salmonella species,
Listeria monocvtogenes, and Staphvlococcus aureus. The inoculated Ioods were held at 30
o
C (to simulate
temperature abuse) and pathogen numbers were monitored over a 24 hour period.
We Iound that
There was no signiIicant increase in pathogen numbers aIter 4 hours storage at 30
o
C.
ChaIIenge test Predictive modeIs
Results oI pathogen growth Irom the challenge tests were checked against those derived Irom predictive
growth models. We Iound that the models predicted a worse outcome than was observed by experimentation
in most instances, but predicted a better outcome than observed in some situations. On that basis, the models
may only be used, with caution.
AppIication of Predictive modeIs of microbiaI growth to deIivery data
We attempted to predict pathogen growth in some home delivered Ioods using the selected model. The
Ioods selected (see Table 1) were characterised by either long delivery times and/or high delivery
temperatures.

Table 1: Foods selected for prediction of microbial growth
Food Delivery time (mins) Final temperature
o
C
Pie 225 19.8
Deli meat (Strasbourg) 200 20.2
Deli meat (Strasbourg) 65 19.9
Milk 215 17.1

We Iound that
1. II the lag times observed in the challenge trials were included in the model predictions, no growth or
growth below detection capacity oI current enumeration methods was predicted.
2. II we assumed that there was no lag time (worst case scenario), then signiIicant growth (deIined as a
10 Iold increase in numbers) was predicted Ior Bacillus cereus and Listeria monocvtogenes.
1

NOTE: As a rule, bacterial Iood poisoning or intoxication Iollows the consumption oI Iood
containing large numbers (greater than 100,000 cIu per gram) oI a pathogen. Thus, while a
signiIicant increase in bacterial numbers may not result in Iood poisoning; it implies an increase in
the risk oI it occurring.
3. No growth oI any oI the 4 pathogens was predicted Ior Ioods that were reIrigerated post-check-out
or were stored in 'Cold Packs during the delivery process even when the delivery time was
extrapolated to 24 hours (assuming that the temperatures oI the Ioods remained at the last recorded
level).

1
The model Ior growth oI B. cereus was Iound to be a good predictor. The model Ior growth oI L. monocvtogenes was
Iound to over-predict growth.

MDU PHL


ConcIusions and Recommendations
Our results indicate that
1. The conditions under which Ioods are normally handled and delivered to homes by the 9 stores
in this study do not appear to increase the Iood saIety risk Ior the majority oI Ioods. However,
abnormal situations such as when milk was leIt out on the door step, because the householder
was not home can adversely aIIect this risk.
2. In a small number oI Ioods, the temperature abuse suIIered during non-reIrigerated delivery may
result in a signiIicant increase in the numbers oI pathogens (iI present).
3. As only a minor part oI the delivery time is spent in the vehicle and a large proportion oI the
delivery time is spent in the store, storage and handling in the store is important.
Stores should attempt to limit heat gain in the Ioods. This can be achieved by appropriate
packing, or better still by reIrigeration or the use oI 'Cold Packs.
Stores that do not reIrigerate the Ioods aIter the check-out, should locate their storage
trolleys in the coolest part oI the store. They should not be stored alongside windows that
are exposed to the sun or near heat generating sources.
Stores should also explore ways oI reducing the delivery times iI Iood is leIt to stand at room
temperature. They should aim to limit delivery times to no more than 4 hours.
4. ReIrigerated delivery vehicles are not necessary iI stores are able to ensure that
Foods are kept as cool as possible, and
The time spent on road transport is maintained at approximately hour.
5. Both reIrigeration and the use oI 'Cold Packs were eIIective in limiting temperature rises in
home deliveries. Using these systems, the time Irom purchase to delivery to the customer can be
extended to 24 hours iI the Iinal temperature does not exceed 10
o
C. However, the time spent on
the road should be kept to a minimum, at around hour.

MDU PHL

Introduction
Supermarkets oIIer home delivery Ior goods purchased in their stores. Currently, this service is largely used
by elderly customers who either do not have a car or who are unable to handle the shopping load. Demand
Ior this service is expected to increase with the projected increase in the number oI elderly people in the
population and the desire by government to encourage them to remain in the community Ior as long as is
practicable. This service may also appeal to others who, Ior liIestyle reasons may wish to avail themselves
oI this convenience.

Home delivery oI supermarket Ioods is managed by the use oI subcontracted couriers who pick up the Iood
and deliver to designated areas. Apart Irom packing the Ioods according to whether they are chilled, Irozen
or shelI-stable, there is normally no attempt at chilling the Ioods Irom the time that they are picked up Irom
the shelI to the time they are delivered to the home. In some instances, the Iood may sit outside Ior some
time iI the customer is not home when the Iood is delivered.

Currently, some municipal councils require perishable Iood to be delivered in reIrigerated vehicles while
others do not. This study will attempt to assess the risks posed by possible temperature abuse during home
delivery and provide decision makers with the scientiIic basis Ior imposing requirements on the conditions
Ior the home delivery oI perishable Ioods that are commensurate with the Iood saIety risks.

This study responds to the supermarket operators` desire to have uniIorm requirements across all
jurisdictions and to health authorities` need Ior evidence-based legislation.
Collaborators
The Microbiological Diagnostic Unit was Iunded by the Victorian Department oI Human Services to conduct
this study. However, this is a multi-Iaceted project and required support Irom, and collaboration with, many
agencies and individuals.
StaII within the Food SaIety Unit oI the Victorian Department oI Human Services provided us with
the initial points oI contacts Ior the Iield work and continuing support throughout the project.
The supermarket chains (Coles and Woolworths/SaIeway) provided us with access to their stores and
relevant staII. The chains donated the Ioods 'purchased Irom their stores and waived delivery
charges.
Various individuals organised their days so that the Ioods could be delivered to them at their homes.
StaII oI the health departments oI several Victorian municipalities were involved to varying degrees
with the conduct oI the supermarket study
Kerry Bell, Queensland Health Department and Tracy Ward, Northern Territory Health assisted with
the interstate temperature monitoring oI home deliveries in Cairns and Casuarina.
Dr Tom Ross oI the University oI Tasmania assessed the suitability oI existing microbial growth
models Ior predicting microbial growth under home delivery conditions.
MDU PHL

Objectives
The objectives oI this project are:
To determine the temperature proIiles oI some reIrigerated ready-to-eat Ioods under diIIerent
climatic, packing and non-reIrigerated delivery conditions.
To determine the potential Ior growth oI pathogenic bacteria that are oI concern under those
conditions, and
To assess the suitability oI existing microbial growth models to predict bacterial growth under home
delivery conditions.
Project outcomes
The Iindings oI this study will:
Provide health authorities with data to aid the Iormulation oI a Iood saIety plan Ior the delivery oI
chilled Ioods to private homes.
Contribute to the development oI uniIorm transport requirements Ior home delivery across all
jurisdictions.
Study Plan
Stage 1:
Establishing collaborative partnerships and method development.
Stage 2:
The supermarket study: Select Ioods in consultation with the supermarkets. Collect temperature proIiles
Irom the time oI Iood selection in the store to the time oI receipt at home. Study conducted in Victoria,
Queensland and the Northern Territory.
Stage 3:
Conduct challenge tests.
Stage 4:
Check results oI pathogen growth obtained Irom the challenge tests Irom Stage 3 against those derived Irom
predictive growth models. Predict pathogen growth in the delivered Ioods by applying the temperature
proIiles obtained Irom Stage 2 to the selected model.
MDU PHL

1. EstabIishing partnerships and method deveIopment
1.1 Estabishing partnerships
This project required the participation oI local governments in Victoria, Queensland and the Northern
Territory, two supermarket chains and householders.

The Food SaIety Unit provided us with the names oI contacts in the supermarket chains and local
government in Queensland and the Northern Territory. MDU PHL staII met with the persons responsible Ior
Iood saIety and quality in each oI the two supermarket chains (Coles and Woolworths/SaIeway) to explain
the project and invite them to participate. We also sought their advice on what Ioods should be included in
the study and requested their assistance with store selection and their Iacilitation with working with the store
managers.

In Melbourne, the relevant local government health department was notiIied oI the project, as a matter oI
courtesy, once the stores were selected. Householders were then recruited, through personal contacts oI
MDU PHL staII and colleagues, to receive the delivered Ioods. The logistics oI store and householder
selection was leIt to the discretion oI the environmental health oIIicers in Queensland and the Northern
Territory.

The assessment oI predictive growth models using data generated Irom the challenge tests in Stage 3 was
sub-contracted to Dr Tom Ross, a consultant to the World Health Organisation on microbial risk assessments
and, a Senior Lecturer at the University oI Tasmania.
1.2 Method development
Various temperature probes were assessed Ior their suitability in use and a pilot run was conducted to test the
operational aspects oI the project. The initial plan to monitor the temperature at the surIace oI the Iood was
shelved aIter we encountered Iunctional and operational diIIiculties in laboratory trials. Laboratory trials
indicated that there was little diIIerence between the core and surIace temperature readings oI the Ioods
selected Ior this project.

MDU PHL


Figure 1: Core and surface Temperature profiles for a fish portion
Green core
Red surIace

Fish core/surface
12h 18h 24We 06h
Time (starting 23/07/2002 11:55:27)
0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
T
e
m
p
e
r
a
t
u
r
e

C

Very little diIIerence between the core and surIace temperatures oI the Ioods were observed. This observation together with the
diIIiculty in securing surIace temperature probes on the Ioods led us to opt Ior the measurement oI core temperatures in this study.
Time on the x-axis is recorded using a 24 hour clock. In this Iigure, recording commenced just beIore 12 noon on 23/7/2002, and
stopped about 10am on 24/7/2002.
MDU PHL

2. Supermarket study - Temperature profiIes of home deIivered
foods
2.1 Study period
The study was carried out between February and March in Victoria, in June in Queensland and in October
2002 in the Northern Territory.
2.2 Sample collection
Nine (9) stores six in Victoria, two in Queensland and one in the Northern Territory were selected Ior this
study. The six Victorian stores (one each with low, medium and high home delivery orders per supermarket
chain) were limited to the Melbourne metropolitan area. The choice oI the interstate stores was leIt to the
discretion oI the local environmental health oIIicer involved.

Milk, minced meat, sliced deli meat, quiche (or pie), ricotta cheese, raw Iish Iillet, cut Iruit pieces and Irozen
peas were selected Ior this study aIter consultation with the supermarkets. These items were chosen because
they were considered to be representative oI the Ioods that were purchased by the majority oI people who
would use the home delivery service that is, senior members oI the community who lived on their own or
with their partner. Frozen peas were included in the purchase even though they required cooking prior to
consumption and were not 'ready to eat Iood because they were commonly purchased by this customer
group. Their inclusion in the shopping bag could potentially keep the temperature oI the other items low as
the Irozen peas could act as a deIacto ice-pack.

Foods were purchased, in quantities appropriate Ior a 2-person household, Irom the selected supermarket by
the project oIIicer (MDU PHL staII in Victoria; local government environmental health oIIicer, interstate) on
consecutive days in a week. Where the designated item was not available, substitutes that closely matched
the Iood, in size and consistency, were purchased.

Shopping times were varied in an attempt to randomise the handling oI the Iood but the extent oI the
variation was limited by the time constraints placed by the store on the acceptance times Ior home deliveries.
In addition, the local daily maximum temperatures Ior each oI the shopping days were obtained Irom the
Bureau oI Meteorology.
2.3 Collection of temperature profiles
Dataloggers (Tinyview Plus Temp, Temperature range -30
o
C 50
o
C, Gemini Data Loggers, Australia
http://www.geminidataloggers.com), Iitted with stainless steel probes were assigned an identiIication code
and calibrated. They were attached to each Iood item by the project oIIicer (see Figure 2) immediately aIter
removal Irom the shelI or at the time oI packaging by the service attendant in the 'Deli section oI the store.
The time oI datalogger attachment was noted Ior each Iood item and householders were asked to note the
time oI each delivery. The dataloggers were retrieved Irom the householders and the temperature proIile
(taken at 5 minute intervals) oI each Iood was downloaded and checked aIter each delivery.
MDU PHL


Figure 2: Attachment of dataloggers to foods

Temperature probes were inserted into all reIrigerated Ioods. In the case oI Irozen peas, the probe was inserted into the package, but
not the Irozen pea. The temperature proIiles are downloaded onto the computer and can be viewed graphically, as shown in the
background oI the photograph oI the milk bottle.
2.4 Home delivery arrangement
Home delivery was requested as the Ioods were processed through the normal check-outs. Store managers
were requested to treat them like any other purchase.

Deliveries were carried out using un-reIrigerated vehicles and the drivers were not inIormed oI the study
being undertaken. In the Victorian study the distance between the stores and the homes Ior delivery was
between 1 and 3 kilometres. The volumes oI, and the distances Ior deliveries are not known Ior the
interstate stores.
MDU PHL

2.5 Findings
2.5.1 DeIivery times
The deliveries were made to householders who lived within the store`s delivery area. For the purpose oI this
study, the delivery time was taken as the time between removal Irom the retail shelI and the time oI receipt in
the home. Thus the delivery time included the
time spent shopping Ior other items and going through the check-out
time the Iood is leIt on the supermarket trolley awaiting delivery plus
time Ior the actual delivery.

The delivery times Ior purchases made at Melbourne and interstate stores are summarised below. Two oI the
twenty nine Melbourne purchases made were not delivered.

Table 2: Delivery times for all stores
Time (minutes)
Minimum Maximum Median
Melbourne stores
1

25 325 110
Interstate stores
2
30 385 150
1
: Calculated Irom 27 deliveries
2
: Calculated Irom 15 deliveries

While there did not appear to be any correlation between the median and maximum delivery times and the
total number oI weekly deliveries handled by the Melbourne stores, minimum delivery times tended to
increase with the number oI deliveries.

Table 3: Delivery times` compared with number of home delivery
orders (Melbourne stores)
Time (minutes) Number of home delivery
orders/week
31 25 285 150
35 50 190 92.5
55 30 240 80
140 50 325 110
186 60 180 135
900 85 230 185
* Does not include times when the deliveries were Iorgotten

Apart Irom diIIerences in individual store practice, the single most important contributing Iactor is the time
oI Iood purchase. Early morning purchases tended to result in longer delivery times. A possible
explanation may be that deliveries are typically scheduled to commence Irom mid-morning and there is thus
a 'compulsory holding period in the store Ior early morning purchases.

MDU PHL

The time spent during transport was Iound to be approximately 30 minutes, which coincides with the
minimum time recorded Ior delivery. The rest oI the time is spent on storage shelves in the store.

The volumes oI deliveries are not known Ior interstate stores, but delivery times appeared to be dependent on
individual store practice.

Table 4: Delivery times for Queensland and Northern Territory stores
Time (minutes) Store
A 130 385 250
B 40 170 145
C 30 225 167.5

The observed median delivery times Ior the interstate stores were longer than those Ior Melbourne stores.
2.5.2 Temperature profiIes
Rough handling oI Iood packages resulted in the dislodgment oI the probe Irom thin parcels such as sliced
meats on several occasions while Irozen peas presented a constant problem. Movement oI the probe in the
bag oI Iree Ilowing peas (we were unable to pierce the Irozen vegetable with the probe) resulted in erratic
temperature readings (Figure 3). For most Ioods, however, the core temperature was adequately monitored
by inserting the probe in the Iood.

Figure 3: Temperature profiles of frozen peas
Supermarket B - peas
-14
-10
-6
-2
2
6
10
0 20 40 60 80 100 120 140 160
Time from SheIf RemovaI to DeIivery (minutes)
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Monday
Tuesday
Wednesday
Thursday
Friday
0

The movement oI the probe in the pack oI Iree Ilowing peas is taken to have caused the highly
variable temperature readings. The trend is, however, Ior the temperature to rise over time.

MDU PHL


Table 5 is a summary oI the Iood temperatures at the time oI receipt in the home. The median delivery
temperature Ior all Ioods was below 13
o
C, although very high temperatures (19-22
o
C) were recorded in a
small number.

Table 5: Summary of Temperature data for foods at point of delivery
Temperature reading at delivery
o
C Food Store Location
Melbourne 0.1 15.7 5.4 Cheese
Qld/NT 4.6 13.1 6.9
Melbourne -1.7 22.7 8.3 Fish
Qld/NT -1.2 14.8 9.3
Melbourne 2.1 19.2 12.15 Fruit
Qld/NT 5.4 17.1 12.6
Melbourne 0.1 17.1 6.9 Milk
Qld/NT 3.9 16.2 10.1
Melbourne 1.5 17.7 8.3 Mince
Qld/NT 5.0 15.1 8.4
Melbourne -14.8 8.5 -3 Peas
Qld/NT -7.8 1.8 -3.2
Melbourne -1.4 19.9 8.8 Pie
Qld/NT -2.7 14.0 8.3
Melbourne 2.3 20.2 9.6 Sliced meats
Qld/NT 7.5 21.1 9.8

Greater temperature increases were observed in Ioods delivered in Melbourne when compared to interstate
deliveries (Table 6).

MDU PHL


Table 6: Temperature increase in foods during home delivery
Temperature increase from shelf selection to delivery
o
C Food Store Location
Melbourne 0.0 13.9 4.4 Cheese
Qld/NT 1.8 6.3 4.0
Melbourne 0.3 24.1 9.0 Fish
Qld/NT 0.5 10.5 7.1
Melbourne 0.3 11.1 5.0 Fruit
Qld/NT 1.9 10.9 4.3
Melbourne 3.0 13.4 6.3 Milk
Qld/NT 2.6 9.6 4.1
Melbourne 1.1 11.4 5.0 Mince
Qld/NT 1.3 7.8 2.8
Melbourne 3.0 20.5 7.8 Peas
Qld/NT 1.5 17.9 5.8
Melbourne 2.9 15.3 6.2 Pie
Qld/NT 1.5 11.1 5.1
Melbourne 3.5 15.6 6.9 Sliced meats
Qld/NT 1.7 11.5 4.8

In summary,
Delivery times
The median delivery times Ior the interstate stores were longer than those Ior Melbourne stores
(Table 2).
Delivery temperatures
The minimum temperatures were generally lower in Melbourne but the median temperatures oI
delivered Iood (other than milk) were very similar Ior Melbourne and the interstate stores (Table 5).
The temperature increase during delivery was greater Ior Melbourne than Ior interstate stores (Table
6).

MDU PHL

2.5.3 Impact of food hoIding after sheIf seIection on temperature profiIes
Three systems Ior the management oI Ioods intended Ior home delivery were observed. The Ioods were held
at
Room temperature
ReIrigerated post check-out, or
In 'Cold Packs.
Room temperature storage post check-out
In most stores, Ioods were leIt at room temperature aIter the check-out. For these Ioods,
The longer the delivery time, the greater the temperature increase observed. Figure 4 demonstrates
this observation.
The rise in temperature was greater when the outside air temperature was higher. (Data not shown)
Larger temperature rises were recorded Ior this group. (Data not shown)
The temperature increase was greater Ior smaller Iood portions. (Data not shown)
A combination oI hot days and long delivery times can result in very large temperature increases. In one
instance, the temperature oI the Iish rose Irom -1.4
o
C to 22.7
o
C (a rise oI 24.1
o
C); the result oI a
combination oI a delivery duration oI 170 minutes and an ambient temperature oI 33
o
C

day on a small Iood
portion (Table 7 on page 20).

Figure 4: Typical food temperature profiles during the ~home delivery
- room temperature storage post check-out
Supermarket A - miIk
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Tuesday
Wednesday
Thursday
Friday

This proIile is typical Ior home delivered Ioods that were not reIrigerated aIter purchase.
The longer the delivery time, the greater the temperature increase.

MDU PHL


In-store reIrigeration post check-out
The temperature proIiles oI Ioods where there was in-store reIrigeration post check-out had a typical stepped
appearance (Figure 5) as a result oI the diIIerent conditions oI storage and handling. However, this pattern
oI temperature change was not observed when Ioods were delivered soon aIter purchase (Thursday proIile in
Figure 5), because oI the limited time spent in reIrigerated storage aIter the check-out.

The beneIit oI reIrigeration was only evident approximately 40 minutes to 1 hour into the delivery time.
Much lower temperature rises were observed in Ioods purchased Irom these stores.

Figure 5: Typical food temperature profiles during the ~home delivery -
refrigerated storage post check-out
Supermarket D - fruit
0
1
2
3
4
5
6
7
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Monday
Tuesday
Wednesday
Thursday
Friday

This proIile is typical Ior home delivered Ioods that were reIrigerated aIter purchase. The
'steps correspond to the storage conditions Irom the time oI removal Irom the shelI (no reIrigeration)
through to reIrigerated storage and Iinally, during road transport (no reIrigeration).

'Cold Pack system
The 'Cold Pack system required Iood to be placed in insulated bags with Ireezer bricks Irom the time it is
removed Irom the shelI to delivery to the householder. Using this system, only small increases, and in some
cases, a reduction in the temperature oI the Iood was observed at the point oI delivery. The temperature
proIiles oI Iood delivered Irom a store that trialled the use oI 'Cold Packs Ior 2 oI the 5 days` deliveries can
be seen in Figure 6 below. Foods were placed directly into the supermarket trolley on the other 3 days.

MDU PHL


Figure 6: Typical food temperature profiles during the ~home delivery
- ~Cold Pack system
Supermarket C - Fish
0
2
4
6
8
10
12
14
16
18
20
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Normal Del A
Normal Del B
Normal Del C
Cold Pack Del A
Cold Pack Del B

This Iigure shows the impact oI the use oI 'Cold Pack delivery Ior Iish. Temperatures rose steadily
in 'normal unreIrigerated in-store storage but Iell over the delivery time with the use oI cold packs.

The relative merits oI the three systems are clearly demonstrated in Figure 7 and Table 7 below.

Figure 7: Impact of the three systems of in-store handling on food temperature
Impact of Temperature ControI
-4
-2
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Post purchase refrigeration
Cold pack
No refrigeration

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Table 7: Temperature increase in selected foods over the delivery period
Food Delivery time
(mins)
Temperature rise
o
C
Max day
temperature
o
C
Quiche
1
225 11.4 36.2
Fish 1
1
170 24.1 33.6
Fish 2
2
315 4.7 23
Fish 3
3
275 5.9 19.2
Fish 4
3
170 4.1 35
Fish 5
3
380 8.9 24
Sliced meat
1
180 7.7 20.9
Sliced meat
1
200 15.6 22.1

1
normal (unreIigerated) system
2
Cold Pack system
3
in store reIrigeration system
Very high temperature increases were recorded in Ioods when there was no in-store reIrigeration Ior
Ioods awaiting delivery (normal system). Temperature increase was minimized with in store reIrigeration
and the 'Cold Pack systems.

MDU PHL

3. ChaIIenge Tests
Retail Ioods are not normally contaminated with pathogens and their growth and survival in these products
cannot be elicited by routine testing. Instead, data on bacterial growth in a Iood system must be generated
using microbiological challenge tests in which the organism oI interest is inoculated into the Iood. Such tests
take into account Iactors such as Iood Iormulation, storage conditions and the eIIect oI competing micro-
organisms in the Iood. They provide a more realistic evaluation oI the potential Ior bacterial growth.

The main concern in this study is the impact oI temperature abuse (presumed to be encountered during home
delivery) on pathogen growth in ready to eat Ioods. The temperature proIiles generated Irom the
supermarket study were used as a guide to the pattern oI 'temperature abuse. The challenge tests were then
carried out under conditions that exceeded even the worst case scenario.

Five diIIerent Iood types, with two examples oI each, were chosen Ior challenge testing. The Ioods selected
were:

Cheese Ricotta and cottage
Sliced meats Devon and strasbourg
Savoury pastry Quiche Lorraine and Chicken & vegetable pie
Fruit Honeydew melon and cantaloupe
Milk Skim and Full cream milk

A cocktail oI 3 strains oI each oI the selected bacterial pathogens, Bacillus cereus, Listeria monocvtogenes,
Staphvlococcus aureus and Salmonella species were used as the challenge organisms. Wherever possible,
we endeavoured to include a strain that had been isolated Irom a similar or a closely related Iood, a Iood
poisoning strain and a strain Irom a recognised culture collection in the cocktail (see Table 8 Ior details).

Table 8: Bacterial strains used in the Challenge tests
Bacterial pathogen Challenge organisms
Bacillus cereus Food poisoning strain Irom rice, ATCC 11778, ATCC13061
Listeria monocvtogenes Strains Irom ice-cream, honeydew melon, sliced meat, human case
Staphvlococcus aureus Strains Irom cheese, human case, NCTC 6571

Salmonella species
S. Anatum var 15 (Irom ice-cream), S. Typhimurium PT135a (Irom
oranges), S. BovismorbiIicans (Irom cattle), S. Typhimurium PT1
(Irom human case), S. SalIord IMVS 1710

The challenge organisms were grown in Brain Heart InIusion broth (Oxoid Ltd., UK) at 37
o
C Ior 24 hours
prior to inoculation into the chilled Ioods (4
o
C). The inoculated Ioods were then held at 30
o
C Ior the
duration oI the challenge test (24 hours) and their bacterial numbers were determined at 0, 2, 4, 6 and 24
hours. The pH and water activity oI the inoculated Ioods were also monitored. The Standard Plate Count
(37
o
C/48 hours) was also determined, over the 24 hour period, Ior the uninoculated Iood.

MDU PHL

Storing Iood in a 30
o
C incubator is equivalent to storing it in a room with an air temperature oI 30
o
C. As
most supermarkets operate at about 20
o
C, this challenge environment can be taken to represent the worst
case scenario. The temperature oI the Iood was monitored during the challenge tests. Figure 8 is typical oI
the temperature proIiles generated during the challenge tests.

Figure 8: Challenge test - Temperature profile of Cottage Cheese
Temperature ProfiIe - Cottage Cheese ChaIIenge Test
0
5
10
15
20
25
30
0 20 40 60 80 100 120 140 160 180 200 220
Time (mins)
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)

There were many diIIerent temperature proIiles Ior Ioods delivered to the homes. The general shape oI the
temperature proIiles generated in the challenge tests (Figure 8) were similar to those observed during home
delivery (Figure 9). However, the temperature increased at a Iaster rate Ior the challenge test.

3.1 Results of Challenge tests
In brieI,
No growth or minimal growth
2
oI pathogenic bacteria was observed in the Iirst 2 to 4 hours oI
storage oI the Ioods at 30
o
C.
Increased pathogen levels were observed aIter 6 hours at 30
o
C Ior milk, Iruit, sliced meats,
quiche/pie.

Figure 10 is typical oI the results observed.

2
Minimal growth is deIined as growth resulting in less than a 10 Iold increase in numbers
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Figure 9: Home Delivery of fish - Temperature profile
Supermarket A - fish
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Tuesday
Wednesday
Thursday
Friday

Temperature proIiles oI the Iish Iillets purchased Irom a supermarket on diIIerent days showing the
trend Ior the temperature to increase with time aIter removal Irom the deli show case. While the
general shape oI the curves are similar to those observed in the challenge tests, the temperature
increased at a Iaster rate during the challenge tests. Thus the time to a 10
o
C increase was approximately
70 minutes Ior Wednesday (steepest temperature increase) while the same temperature rise was recorded
in less than 40 minutes Ior Ioods in the challenge tests.

Figure 10: Challenge test - Pie
ChaIIenge Test - Pie
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

The above Iigure is typical oI the patterns observed in the challenge tests. A period oI no growth
(lag phase) is Iollowed by exponential growth during which, the bacteria double in numbers at
regular intervals. The numbers level out aIter a period arriving at what is known as the stationary phase.
MDU PHL

4. ChaIIenge tests resuIts vs Predictive modeIs
The results oI the challenge tests were compared to those generated by mathematical models that predict
microbial growth in Ioods. Comparisons were conducted Ior Iour pathogens (S. aureus, B. cereus,
Salmonella species and L. monocvtogenes). Models were not readily available in the public domain Ior all
oI these organisms, and some models had to be created Ior this project using existing published data.

The models selected Ior the Iour pathogens were:
S. aureus Ross & McMeekin (1991), modiIied by the inclusion oI an empirical term Ior
pH, based on the pH term introduced by Presser et al (1997)
L. monocvtogenes Unpublished model developed at the University oI Tasmania and based on
studies by Ross (1993) and Tienungoon (1998)
B. cereus Model developed using the data oI Benedict et al (1993), Iitted to a square root
type model with terms Ior temperature, water activity and pH using UltraIit 3.0
soItware (BiosoIt Corporation, Cambridge, UK)
Salmonella spp. Model developed Irom data oI Gibson et al (1988) in a similar manner to the
B. cereus model.
4.1 Methods of Comparison
Two methods oI evaluation oI the perIormance oI the models were used.
1. Qualitative comparison oI the observed pathogen growth curve Irom the challenge trial and the
predicted growth curve generated by the model.
2. Comparison oI the exponential growth rate.
3

4.1.1 QuaIitative comparison
Using this method, the model`s growth predictions are compared with the observed growth oI the challenge
organism. The qualitative comparison Ior B. cereus growth in Figure 11 indicates that there is good
agreement between the model prediction and the observed growth oI the challenge organism in melon. The
Iigure also shows the change in Standard Plate Count (SPC) and the change in temperatures.

Results oI the qualitative comparisons Ior the various Iood:pathogen combinations are summarised in Table
9.

3
The exponential growth rate is estimated Irom the challenge test data. While not all datasets allowed reliable
estimation oI the exponential growth rate, it was noted that in almost all the challenge trial datasets, the 6 hour samples
did represent cultures in exponential growth and the 24 hour samples were clearly indicative oI cultures in stationary
phase.

MDU PHL


Figure 11: Qualitative evaluation of a predictive model for the growth of B. cereus
compared to observations from a challenge trial in melon involving B. cereus.

Table 9: Fit of the predictive growth model to Challenge test data - Qualitative assessment
Model Performance in predicting growth in the different foods Pathogen
Good fit` Over-predicted Under-predicted
S. aureus Cottage cheese, Devon,
strassbourg, melon,
quiche, skim milk
Ricotta Cantaloupe, Iull cream
milk, pie
L. monocvtogenes Devon, skim milk,
strassbourg, melon
Cantaloupe, cottage
cheese, Iull cream
milk, pie, quiche,
ricotta

B. cereus Cantaloupe, cottage
cheese, Devon, Iull
cream milk, melon, pie,
quiche, skim milk,
strassbourg
Ricotta
Salmonella spp. Melon, pie, quiche,
ricotta, skim milk
Cantaloupe, cottage
cheese, Iull cream milk
Devon, strassbourg
*the Iit is rated as good iI the growth rates are comparable and the combination oI the lag period and growth rate do not result in the
under-estimation oI the growth oI the pathogen.

In summary, the models predicted a worse outcome than was observed by experimentation in most instances,
but predicted a better outcome than observed in some situations. On that basis, the models may only be
used, with caution, to predict bacterial growth in the Ioods tested.
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4.2 Comparison of Exponential Growth Rates.
The evaluation oI predictive model perIormance is usually achieved by comparison oI the model`s growth
rate estimates to experimentally derived estimates oI growth rates in Ioods. Two established indices oI
model perIormance are the bias Iactor (B
f
) and accuracy or deviance Iactor (A
f
).

The bias Iactor is a measure oI how the model, on average, over- or under-predicts.
A bias Iactor oI 1.1 Ior response time (e.g. generation time, lag time, time Ior a ten-Iold increase etc.)
indicates that the model is Iail-dangerous` because it predicts on average, a 10 longer generation
time than are observed.
A bias Iactor less than one Ior generation time indicates that a model is, in general, Iail-saIe`.
A bias Iactor oI 0.5 indicates a poor model that is overly conservative because it predicts generation
times, on average, halI oI that actually observed.
II growth rates are compared, the interpretation is reversed.

The accuracy Iactor is a measure oI the spread oI results about the prediction. The larger the value, the less
accurate is the average estimate. Thus, an accuracy Iactor oI two indicates that the prediction is, on average,
a Iactor oI two diIIerent Irom the observed value i.e. either halI as large or twice as large, while a value oI
one indicates that there is perIect agreement between all predicted and observed values.

The bias and accuracy Iactors were calculated using comparisons between the measurements oI growth Ior
challenge organisms Irom 4 and 6 hours aIter the start oI the challenge tests and the respective growth
predictions Irom the models (Table 10).

Table 10: Calculated Bias and Accuracy Factors for growth rates for the Models used in
this Study Compared to the Challenge Trial data
Organism Bias Factor
#
Accuracy Factor
Salmonella 0.96 1.47
L. monocvtogenes 1.98 1.85
B. cereus 1.30 1.44
S. aureus 1.03 1.40
#
a bias Iactor greater than 1 indicates that the model overpredicts the observed growth rate and is 'Iail-saIe
On the whole, the models Ior S. aureus, B. cereus and Salmonella species provided good or Iair estimates oI
growth rate oI the challenge organisms, although predictions Ior individual Iood:pathogen combinations
were less reliable.
MDU PHL


As a rule oI thumb, Ross et al., (2000) suggested that Ior every Iactor in a model, the accuracy Iactor would
increase by 0.10 to 0.15. Thus, according to Ross et al, accuracy Iactors in the range 1.30 to 1.45 would be
considered acceptable Ior three parameter models as used in this study. The results suggest that,
The Salmonella and S. aureus models are particularly good` when both the bias and the accuracy
Iactors are taken into account.
The combination oI the large bias Iactor and large accuracy Iactor identiIies the L. monocvtogenes
model as being overly conservative, i.e. it predicts much Iaster growth oI the pathogen than was
actually observed Ior all the examples. This implies that the true potential growth oI L.
monocvtogenes in the delivery period (Table 11) may be overestimated by nearly 100.
4.3 Application of Predictive models of microbial growth to delivery data
The time/temperature conditions observed in the supermarket trials were applied to the mathematical models
to estimate the impact on the growth oI pathogens in the home delivered Ioods. Two sets oI estimates were
made, i.e. one with, and the other without, the inclusion oI the lag times
4
observed in the challenge tests.

The choice oI the estimate depends on whether one would expect pathogens, iI present in Ioods, to display a
lag time beIore initiating growth when temperatures rise to levels permitting their growth.
Psychrotrophs
5
such as L. monocvtogenes and some strains oI B. cereus would most likely be
resolving their lag times during reIrigerated storage. Whether the lag times are completely resolved
beIore the commencement oI home delivery is unknown. Growth oI these bacteria are expected to
proceed with no or relatively short lag times.
While it is not clear iI the growth oI mesophilic
6
pathogens in reIrigerated Ioods would involve a lag
time, it is noted that lag times equivalent to Iour to six generations oI growth
7
have been observed in
many studies (Ross, 1999). As lag times are not resolved under conditions that prevent growth, it is
reasonable to assume that any growth oI mesophilic pathogens during home delivery would be
preceded by a lag time.
4.3.1 Predicting growth in home deIivered foods
The models were used to predict pathogenic bacterial growth in the delivered Ioods that were thought to have
undergone the most temperature abuse (based on the time taken Ior the delivery and the temperature at the
point oI delivery). Under the delivery conditions observed in this study,
No growth or growth less than detectable using existing enumeration methods was predicted iI the
lag times observed in the challenge tests were included in the model predictions.
SigniIicant amounts oI growth were predicted in some instances (as shown in Table 11) iI there was
no lag period.
Note: Most methods oI microbial enumeration have an error oI the order oI ~0.5 logCFU/g, and predictions oI
growth above this amount might warrant Iurther attention.

4
The lag time is the time that bacteria take to adjust to their environment, beIore multiplication commences
5
bacteria capable oI growing under reIrigeration conditions
6
mesophilic bacteria grow between 10 and 45
o
C
7
a 1 log (10 Iold) increase in bacterial numbers is equivalent to 3.32 generations oI growth
MDU PHL


Table 11: Prediction of pathogen growth under observed delivery conditions for selected foods -
assuming no lag time (worst case scenario)
Predicted increase (Log cfu) Food Del time
(mins)
Delivery
Temp
Salmonella B. cereus S. aureus L. monocytogenes
Pie 225 19.8
o
C 0.78 1.42 0.32 0.83
Deli meat 200 20.2
o
C 0.37 0.98 0.85 0.78
Deli meat 65 19.9
o
C 0.03 0.13 0.05 0.07
Milk 215 17.1
o
C 0.74 1.20 0.31 1.00
SigniIicant growth (particularly oI B. cereus) was observed in the 3 Ioods with extended delivery times. As the no lag assumption is
probably true Ior some strains oI B. cereus, this observation is oI some concern. As a rule, bacterial Iood poisoning or intoxication
Iollows the consumption oI Iood containing large numbers (greater than 100,000 cIu per gram) oI a pathogen or toxins. Thus, while
a signiIicant increase in bacterial numbers may not result in Iood poisoning; it implies an increase in risk oI it occurring.

No growth oI any oI the 4 pathogens was predicted Ior Ioods that were reIrigerated post check-out and Ior
Ioods under the observed 'Cold Pack conditions oI delivery even when the delivery times were extrapolated
to 24 hours. The assumptions made in this projection are that
The reIrigerated Ioods maintain the temperatures they had attained (in the in-store coolroom) prior to
road transport. This temperature is assumed to be the Iood temperature approximately 30 minutes
prior to receipt in the home.
The temperatures oI 'Cold Pack Ioods remained at the last recorded level.
These assumption are reasonable as the time/temperature proIiles oI the Ioods have leveled or, in some
instances, were still Ialling in the reIrigerated Ioods prior to road transport and in the 'Cold Packs Ior the
whole duration oI the delivery.
MDU PHL

5. ConcIusions
Home delivery
1. The duration Ior home delivery (deIined as the time Irom removal oI Iood selection in the store
to the time oI receipt at home) does not truly reIlect the time spent in road transport.
Approximately halI an hour was spent on the road.
2. The normal conditions under which Ioods are handled and delivered to homes by the 9 stores in
this study do not appear to increase the Iood saIety risk Ior the majority oI Ioods. However,
unIoreseen situations such as when milk was leIt out on the door step, because the
householder was not home can aIIect this risk.
3. In a small number oI Ioods, the temperature abuse suIIered during non-reIrigerated delivery
may result in a signiIicant increase in the numbers oI pathogens (iI present).
4. High day temperatures, long delivery times and smaller portions contribute to temperature
increases during home delivery.
5. Melbourne stores tended to have shorter delivery times but greater temperature increases were
observed Ior Ioods delivered in Melbourne.
6. ReIrigerated delivery vehicles are not necessary iI stores are able to ensure that
Foods are kept as cool as possible, and
The time spent on road transport is maintained at approximately hour.

Store practice
7. As only a minor part oI the delivery time is spent in the vehicle and a large proportion oI the
delivery time is spent in the store, storage and handling in the store is important.
Stores should attempt to limit heat gain in the Ioods. This can be achieved by appropriate
packing, or better still by reIrigeration or the use oI 'Cold Packs.
Both reIrigeration and the use oI 'Cold Packs were eIIective in limiting temperature rises in
home deliveries. Using these systems, delivery times can saIely be extended to 24 hours (iI
the Iinal temperature does not exceed 10
o
C).
Stores that do not reIrigerate the Ioods aIter the check-out, should locate their storage
trolleys in the coolest part oI the store. They should not be stored alongside windows that
are exposed to the sun or near heat generating sources.
Stores should also explore ways oI reducing the delivery times iI Iood is leIt to stand at room
temperature. They should aim to limit the total delivery times to no more than 4 hours.
Potential Ior bacterial growth during home delivery
8. The majority oI deliveries involved moderate temperature increases over a short period (median
temperature increase oI less than 13
o
C over a median period oI less than 2 hours Ior Melbourne
stores). No increase in pathogen numbers was demonstrated in our laboratory tests in the Iirst 4
hours when Ioods were subjected to temperature conditions that were more severe than those
expected during normal home delivery
8
. The above observation was corroborated by predictive
modelling oI bacterial growth.

8
Foods were held at 30
o
C during the Challenge tests while stores operate, on average, at about 20
o
C.
MDU PHL

9. A substantial increase in product temperature was observed in a small number oI Ioods during
non- reIrigerated home delivery. This was predicted to give rise to a signiIicant increase in
numbers oI some pathogenic bacteria.

Models Ior predicting bacterial growth
10. The mathematical models predicted a worse outcome than was observed by experimentation in
most instances, but predicted a better outcome than observed in some situations. On that basis,
the models may only be used, with caution, to predict bacterial growth.
MDU PHL

References
Benedict, R.C., Partridge, T., Wells, D. and Buchanan, R.L. (1993). Bacillus cereus: Aerobic growth
kinetics. Journal of Food Protection, 56: 211-214.
Gibson, A.M., Bratchell, B and Roberts, T.A. (1988). Predicting microbial growth: growth responses oI
salmonellae in a laboratory medium as aIIected by pH, spdium chloride and storage temperature.
International Journal of Food Microbiologv, 6: 155 - 178.
Presser, K. Ratkowsky, D.A., and Ross, T (1997). Modelling the growth rate oI Escherichia coli as a Iunction
oI pH and lactic acid concentration. Applied and Environmental Microbiologv, 63:2355-2360.
Ross, T. (1993). A philosophv for the development of kinetic models in predictive microbiologv. PhD Thesis,
University oI Tasmania, Hobart, Australia.
Ross, T. and McMeekin, T.A. (1991) Predictive Microbiology. Applications oI a square root model. Food
Australia, 43: 202-207.
Ross, T, Dalgaard, P, Tienungoon, S. (2000) Predictive modelling oI the growth and survival oI Listeria in
Iishery products`, International Journal of Food Microbiologv, 62: 231-245.
Tienungoon, S. (1998). Some Aspects of the Ecologv of Listeria monocytogenes in Salmonid Aquaculture.
PhD. Thesis, University oI Tasmania, Hobart, Australia.
MDU PHL


Appendices
MDU PHL

A1. Temperature measurement triaIs -comparison of core vs
surface temperature measurements
Experiments were conducted in the laboratory to determine iI the surIace temperatures were very diIIerent
Irom the temperatures at the core (geometric centre) oI the Iood.

The plan to monitor the temperature at the surIace oI the Iood was shelved aIter
We encountered Iunctional and operational diIIiculties when we attempted to securely attach the
temperature probe to the surIace oI the Iood items in our laboratory trials, and
Laboratory trials indicated that there was little diIIerence between the core and surIace temperature
readings oI the Ioods selected Ior this project.
The graphs below summarises our Iindings.

Green core
Red surIace

Fish core/surface
12h 18h 24We 06h
Time (starting 23/07/2002 11:55:27)
0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
T
e
m
p
e
r
a
t
u
r
e

C

Fruit core/surface
12h 18h 24We 06h
Time (starting 23/07/2002 11:35:02)
0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
T
e
m
p
e
r
a
t
u
r
e

C

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Mince core/surface
18h 24We 06h
Time (starting 23/07/2002 12:01:06)
0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
T
e
m
p
e
r
a
t
u
r
e

C

Quiche core/surface
12h 18h 24We 06h
Time (starting 23/07/2002 11:32:24)
-5.0
0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
T
e
m
p
e
r
a
t
u
r
e

C

Ricotta core/surface
18h 24We 06h
Time (starting 23/07/2002 12:04:48)
-5.0
0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
T
e
m
p
e
r
a
t
u
r
e

C

Slice core/surface
12h 18h 24We 06h
Time (starting 23/07/2002 11:51:01)
-40.0
-30.0
-20.0
-10.0
0
10.0
20.0
30.0
40.0
T
e
m
p
e
r
a
t
u
r
e

C

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A2. Temperature profiIes for Home deIiveries
Eight Ioods (cheese, Iish, Iruit, milk, mince, pie, deli meat, peas) were purchased Irom 12 stores over 5
consecutive days and their temperatures were monitored, Irom the time oI selection Irom the shelves to the
time oI receipt in the home. Figures A2.1 to A2.3 are graphic descriptions oI the temperature proIiles Ior
Ioods handled by the three systems (room temperature shelI storage, reIrigerated and 'Cold Pack) aIter the
check-out.

Fig A2.1: Typical food temperature profiles during the ~home delivery - foods stored at room
temperature post check-out

Supermarket A- cheese
0
2
4
6
8
10
12
14
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Tuesday
Wednesday
Thursday
Friday

Supermarket A - fish
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Tuesday
Wednesday
Thursday
Friday

Supermarket A - fruit
4
6
8
10
12
14
16
18
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Tuesday
Wednesday
Thursday
Friday

Supermarket A - miIk
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Tuesday
Wednesday
Thursday
Friday

Supermarket A - mince
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Tuesday
Wednesday
Thursday
Friday

MDU PHL


Fig A2.2: Typical food temperature profiles during the ~home delivery - foods stored refrigerated
post check-out

Supermarket D - cheese
0
1
2
3
4
5
6
7
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
tu
r
e
(o
C
)
Monday 14th
Tuesday 15th
Wednesday 16th
Thursday 17th
Friday 18th

Supermarket D - miIk
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
tu
r
e
(o
C
)
Monday 14th
Tuesday 15th
Wednesday 16th
Thursday 17th
Friday 18th

Supermarket D - fruit
0
1
2
3
4
5
6
7
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
tu
r
e
(o
C
)
Monday
Tuesday
Wednesday
Thursday
Friday

Supermarket D - fish
0
2
4
6
8
10
12
14
16
18
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
tu
r
e
(o
C
)
Monday 14th
Tuesday 15th
Wednesday 16th
Thursday 17th
Friday 18th

Supermarket D - mince
0
1
2
3
4
5
6
7
8
9
10
0 20 40 60 80 100 120 140 160 180 200 220 240 260 280
Time from SheIf RemovaI to DeIovery (minutes)
T
e
m
p
e
r
a
tu
r
e
(o
C
) Monday
Tuesday
Wednesday
Thursday
Friday
MDU PHL


Fig A2.3: Typical food temperature profiles during the ~home delivery - Cold Pack

Supermarket C - Cheese
0
2
4
6
8
10
12
14
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Normal Del A
Normal Del B
Normal Del C
Cold Pack Del A
Cold Pack Del B

Supermarket C - Fish
0
5
10
15
20
25
0 20 40 60 80 100 120 140 160 180 200 220
T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
Normal Del A
Normal Del B
Normal Del C
Cold Pack Del A
Cold Pack Del B

Supermarket C - miIk
0
2
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Normal Del A
Normal Del B
Normal Del C
Cold Pack Del A
Cold Pack Del B

Supermarket C - mince
4
6
8
10
12
14
16
0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Normal Del A
Normal Del B
Normal Del C
Cold Pack Del A
Cold Pack Del B

Supermarket C - fruit
0
2
4
6
8
10
12
14
16
18
0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340
T
e
m
p
e
r
a
t
u
r
e
(
o
C
)
Normal Del A
Normal Del C
Cold Pack Del A
Cold Pack Del B

Note:
'Normal deliveries on 3 days (i.e. Ioods were
leIt unreIrigerated on trolleys while waiting Ior
drivers to deliver to the homes)
'Cold Pack deliveries on 2 days

MDU PHL

A3. ChaIIenge test resuIts
Duplicate sets oI 5 Ioods were challenged a cocktail oI 3 strains oI each oI the selected bacterial
pathogens, Bacillus cereus, Listeria monocvtogenes, Staphvlococcus aureus and Salmonella species.
Wherever possible, we endeavoured to include a strain that had been isolated Irom a similar or a
closely related Iood, a Iood poisoning strain and a strain Irom a recognised culture collection in the
cocktail.
The inoculated Ioods were held at 30
o
C (to simulate temperature abuse) and pathogen numbers were
monitored over a 24 hour period.

Bacterial strains used in the Challenge tests

Bacterial pathogen Challenge organisms
Bacillus cereus Food poisoning strain Irom rice, ATCC 11778, ATCC13061
Listeria monocvtogenes Strains Irom ice-cream, honeydew melon, sliced meat, human case
Staphvlococcus aureus Strains Irom cheese, human case, NCTC 6571

Salmonella species
S. Anatum var 15 (Irom ice-cream), S. Typhimurium PT135a (Irom
oranges), S. BovismorbiIicans (Irom cattle), S. Typhimurium PT1
(Irom human case), S. SalIord IMVS 1710

Results oI challenge tests are tabulated in the Iollowing pages

MDU PHL

A3.1 Ricotta Cheese
Table A3.1.1: Results of Challenge test - B. cereus
Reading after storage at 30
o
C
0 hr 2 hr 4 hr 6 hr 24 hr
Count oI B. cereus 2.9 x
10
3
2.6 x
10
3

7.0 x
10
3

2.0 x
10
4

2.0 x
10
7

pH oI inoculated Iood 6.54 6.69 6.75 6.69 6.28
Colony count oI uninoculated
Iood (37
o
C/48 hours)
1.3 x
10
4

3.9 x
10
3

5.8 x
10
3

5.1 x
10
4

7.8 x
10
7

pH oI uninoculated Iood 6.58 6.72 6.73 6.70 6.75
Water activity (a
w
) oI Iood 0.993 0.993

Table A3.1.2: Results of Challenge test - Salmonella spp.
o
C
Count oI Salmonella spp. 1.0 x 10
4
9.1 x 10
3
1.6 x 10
4
6.7 x 10
4
1.0 x 10
8

Iood (37
o
C/48 hours) 2.5 x 10
4

No
result

9.2 x 10
4

1.0 x 10
6

1.5 x 10
7

Water activity (a
w
) oI Iood 0.990 0.986

Table A3.1.3: Results of Challenge test - L. monocytogenes
o
C
Count oI L. monocvtogenes 3.1 x 10
4
3.1 x 10
4
3.7 x 10
4
1.0 x 10
5
1.1 x 10
7

Iood (37
o
C/48 hours)
1.1 x 10
3

2.2 x 10
2

9.6 x 10
4

1.7 x 10
3

2.4 x 10
7

Water activity (a
w
) oI Iood 0.994 0.998

Table A3.1.4: Results of Challenge test - S. aureus
o
C
Count oI S. aureus 1.4 x 10
4
1.9 x 10
4
5.6 x 10
4
2.2 x 10
5
1.0 x 10
8

Iood (37
o
C/48 hours) 4.0 x 10
3

1.3 x 10
4

No
result

No
result

5.3 x 10
6

Water activity (a
w
) oI Iood 0.993 0.993
MDU PHL

A3.2 Cottage Cheese

o
C
Count oI B. cereus
1.8 x 10
3
2.3 x 10
3
2.6 x 10
3
3.5 x 10
3

No
Result
Iood (37
o
C/48 hours)
1.1 x 10
3

4.0 x 10
2

3.0 x 10
2

7.0 x 10
2

7.9 x 10
5

Water activity (a
w
) oI Iood 0.990 0.990

o
C
4
1.1 x 10
4
1.5 x 10
4
4.2 x 10
4
2.6 x 10
6

Iood (37
o
C/48 hours)
300

300

300

300

6.7 x 10
4

Water activity (a
w
) oI Iood 0.991 0.987

o
C
4
2.7 x 10
4
3.6 x 10
4
4.6 x 10
4
6.4 x 10
5

Iood (37
o
C/48 hours)
300

300

300

300

300

Water activity (a
w
) oI Iood 0.988 0.987

o
C
4
1.2 x 10
4
1.4 x 10
4
2.4 x 10
4
8.4 x 10
6

Iood (37
o
C/48 hours)
8.8 x 10
5

1.4 x 10
5

7.2 x 10
4

1.1 x 10
4

1.1 x 10
4

Water activity (a
w
) oI Iood 0.990 0.998

MDU PHL


Fig A3.1: Challenge Test - Ricotta Cheese
ChaIIenge Test - Ricotta Cheese
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

g
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

FigA 3.2: Challenge test - Cottage cheese

ChaIIenge Test - Cottage Cheese
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

MDU PHL

A3.3 Full cream milk

o
C
Count oI B. cereus 1.3 x 10
4
1.3 x 10
4
3.0 x 10
4
1.4 x 10
5
5.5 x 10
7

pH oI inoculated Iood 6.79 6.87 6.80 6.69 ND
Iood (37
o
C/48 hours)
300

300

300

300

2.5 x 10
7

pH oI uninoculated Iood 6.80 6.84 6.78 6.71 ND
Water activity (a
w
) oI Iood 0.993 0.992

o
C
4
1.3 x 10
4
5.3 x 10
4
1.7 x 10
5
9.2 x 10
8

Iood (37
o
C/48 hours)
300

300

300

300

1.9 x 10
7

Water activity (a
w
) oI Iood 0.993 0.993

o
C
4
2.8 x 10
4
3.1 x 10
4
1.0 x 10
5
8.1 x 10
7

Iood (37
o
C/48 hours)
1.3 x 10
3

1.8 x 10
3

1.8 x 10
3

2.3 x 10
3

3.0 x 10
7

Water activity (a
w
) oI Iood 0.993 0.999

o
C
3
8.7 x 10
3
1.6 x 10
4
1.4 x 10
5
1.2 x 10
8

Iood (37
o
C/48 hours)
9.5 x 10
2

1.1 x 10
3

8.2 x 10
2

7.7 x 10
2

1.2 x 10
7

Water activity (a
w
) oI Iood 0.993 0.995

MDU PHL

A3.4 Skim milk

o
C
4
1.5 x 10
4
1.4 x 10
4
1.4 x 10
5
1.4 x 10
7

pH oI inoculated Iood 6.71 6.81 6.72 6.73 ND
Iood (37
o
C/48 hours)
300

300

300

3.5 x 10
2

1.0 x 10
7

pH oI uninoculated Iood 6.75 6.79 6.69 6.73 ND
Water activity (a
w
) oI Iood 0.992 0.996

o
C
4
1.3 x 10
4
3.6 x 10
4
2.8 x 10
5
8.8 x 10
8

Iood (37
o
C/48 hours)
300

300

300

300

1.4 x 10
7

Water activity (a
w
) oI Iood 0.993 0.993

o
C
4
2.8 x 10
4
2.8 x 10
4
1.2 x 10
5
2.2 x 10
7

Iood (37
o
C/48 hours)
3.0 x 10
2

300

3.5 x 10
2

300

1.6 x 10
8

Water activity (a
w
) oI Iood 0.993 0.990

o
C
3
9.7 x 10
3
8.7 x 10
3
4.5 x 10
4
1.7 x 10
7

Iood (37
o
C/48 hours)
5.0 x 10
2

4.6 x 10
2

4.0 x 10
2

3.5 x 10
2

2.7 x 10
6

Water activity (a
w
) oI Iood 0.991 0.994

MDU PHL

Figure A3.3: Challenge test - Full Cream Milk
ChaIIenge Test - FuII Cream MiIk
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

Figure A3.4: Challenge test - Skim Milk
ChaIIenge Test - Skim MiIk
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

MDU PHL

A3.5 Cantaloupe

o
C
3
2.2 x 10
3
5.6 x 10
3
1.1 x 10
5
2.2 x 10
8

Iood (37
o
C/48 hours)
300

1.8 x 10
3

300

3.6 x 10
3

3.7 x 10
8

Water activity (a
w
) oI Iood 0.992 0.990

o
C
4
1.2 x 10
4
3.6 x 10
4
1.1 x 10
5
4.9 x 10
8

Iood (37
o
C/48 hours)
1.2 x 10
3

2.2 x 10
3

1.9 x 10
3

1.5 x 10
4

3.8 x 10
8

Water activity (a
w
) oI Iood 0.990 0.989

o
C
4
3.2 x 10
4
6.1 x 10
4
2.1 x 10
5
8.4 x 10
7

Iood (37
o
C/48 hours)
7.6 x 10
3

5.7 x 10
4

6.1 x 10
3

1.2 x 10
4

1.1 x 10
9

Water activity (a
w
) oI Iood 0.967 0.988

o
C
4
2.4 x 10
4
5.7 x 10
4
2.2 x 10
5
2.7 x 10
8

Iood (37
o
C/48 hours)
No
result

1.8 x 10
4

7.3 x 10
4

1.2 x 10
7

1.0 x 10
9

Water activity (a
w
) oI Iood 0.991 0.962

MDU PHL

A3.6 Melon

o
C
3
1.8 x 10
3
7.5 x 10
3
1.4 x 10
5
2.0 x 10
7

Iood (37
o
C/48 hours)
300

300

300

4.5 x 10
2

3.2 x 10
7

Water activity (a
w
) oI Iood 0.995 0.993

o
C
3
9.0 x 10
3
2.5 x 10
4
2.8 x 10
5
7.1 x 10
8

Iood (37
o
C/48 hours)
300

300

300

300

3.5 x 10
7

Water activity (a
w
) oI Iood 0.989 0.990

o
C
4
1.9 x 10
4
3.6 x 10
4
2.5 x 10
5
2.5 x 10
8

Iood (37
o
C/48 hours)
1.5 x 10
3

4.9 x 10
2

5.7 x 10
2

4.2 x 10
2

2.8 x 10
7

Water activity (a
w
) oI Iood 0.992 0.990

o
C
4
1.4 x 10
4
4.2 x 10
4
4.7 x 10
5
8.9 x 10
8

Iood (37
o
C/48 hours)
No
result

1.5 x 10
3

8.0 x 10
2

2.7 x 10
3

9.2 x 10
8

Water activity (a
w
) oI Iood 0.991 0.989

MDU PHL

Fig A3.5: Challenge Test - Cantaloupe
ChaIIenge Test - CantaIoupe
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

Fig A3.6: Challenge test - Melon
ChaIIenge Test - MeIon
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

MDU PHL

A3.7 Devon

o
C
3
1.9 x 10
3
2.8 x 10
3
1.2 x 10
4
1.7 x 10
8

Iood (37
o
C/48 hours)
300

300

5.5 x 10
2

1.3 x 10
3

1.0 x 10
7

Water activity (a
w
) oI Iood 0.969 0.969

o
C
4
1.0 x 10
4
1.2 x 10
4
1.7 x 10
5
3.1 x 10
8

Iood (37
o
C/48 hours)
300

300

300

300

1.7 x 10
6

Water activity (a
w
) oI Iood 0.962 0.963

o
C
4
5.8 x 10
4
6.4 x 10
4
1.4 x 10
5
9.5 x 10
8

Iood (37
o
C/48 hours)
5.8 x 10
4

1.3 x 10
5

3.1 x 10
3

4.4 x 10
4

6.6 x 10
7

Water activity (a
w
) oI Iood 0.972 0.969

o
C
4
1.5 x 10
4
6.6 x 10
4
3.8 x 10
5
2.7 x 10
9

Iood (37
o
C/48 hours)
2.1 x 10
5

9.7 x 10
5

1.1 x 10
6

9.5 x 10
5

9.4 x 10
8

Water activity (a
w
) oI Iood 0.973 0.970

MDU PHL

A3.8 Strasbourg

o
C
3
2.2 x 10
3
3.2 x 10
3
2.3 x 10
4
3.4 x 10
8

Iood (37
o
C/48 hours)
300

7.6 x 10
2

7.2 x 10
2

5.4 x 10
3

5.4 x 10
7

Water activity (a
w
) oI Iood 0.973 0.972

o
C
4
1.2 x 10
4
1.7 x 10
4
1.2 x 10
5
1.6 x 10
9

Iood (37
o
C/48 hours)
300 300 300 300
2.4 x 10
7

Water activity (a
w
) oI Iood 0.970 0.970

o
C
4
7.0 x 10
4
5.4 x 10
4
1.0 x 10
5
1.8 x 10
9

Iood (37
o
C/48 hours) 3.0 x 10
3
4.8 x 10
3
8.0 x 10
3
1.9 x 10
4
5.1 x 10
7

Water activity (a
w
) oI Iood 0.971 0.968

o
C
4
1.6 x 10
4
5.4 x 10
4
4.1 x 10
5
3.0 x 10
9

Iood (37
o
C/48 hours) 1.1 x 10
5
2.0 x 10
5
1.8 x 10
5
4.5 x 10
5
4.0 x 10
7

Water activity (a
w
) oI Iood 0.973 0.972

MDU PHL

Fig A3.7: Challenge test - Devon
ChaIIenge Test - Devon
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

Fig A3.8: Challenge test - Strasbourg
ChaIIenge Test - Strasbourg
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

MDU PHL

A3.9 Quiche

o
C
3
3.2 x 10
3
6.2 x 10
3
1.4 x 10
5
1.6 x 10
8

Iood (37
o
C/48 hours) 2.3 x 10
4
3.0 x 10
2
3.4 x 10
3
1.1 x 10
3
1.3 x 10
7

Water activity (a
w
) oI Iood 0.988 0.989

o
C
4
1.4 x 10
4
3.6 x 10
4
1.9 x 10
5
4.6 x 10
8

Iood (37
o
C/48 hours) 300 3.2 x 10
3
9.9 x 10
2
6.1 x 10
3
1.5 x 10
8

Water activity (a
w
) oI Iood 0.986 0.979

o
C
4
2.4 x 10
4
3.1 x 10
4
5.4 x 10
4
2.1 x 10
5

Iood (37
o
C/48 hours) 300 300 300 300 7.2 x 10
7

Water activity (a
w
) oI Iood 0.990 0.997

o
C
4
1.1 x 10
4
1.8 x 10
4
1.1 x 10
5
5.5 x 10
8

Iood (37
o
C/48 hours) 300 300 300 300 1.7 x 10
4

Water activity (a
w
) oI Iood 0.986 0.979

MDU PHL

A3.10 Pie

o
C
3
2.7 x 10
3
1.1 x 10
4
2.1 x 10
5
2.0 x 10
5

Iood (37
o
C/48 hours) 1.2 x 10
2
300 300 4.0 x 10
2
2.5 x 10
5

Water activity (a
w
) oI Iood 0.979 0.989

o
C
4
1.0 x 10
4
2.5 x 10
4
1.8 x 10
5
4.2 x 10
8

Iood (37
o
C/48 hours) 300 300 300 7.0 x 10
2
3.5 x 10
6

Water activity (a
w
) oI Iood 0.985 0.982

o
C
4
2.9 x 10
4
3.1 x 10
4
9.2 x 10
4
3.8 x 10
8

Iood (37
o
C/48 hours) 1.8 x 10
3
2.2 x 10
3
300 2.2 x 10
3
1.3 x 10
8

Water activity (a
w
) oI Iood 0.989 0.995

o
C
4
1.1 x 10
4
5.7 x 10
4
4.1 x 10
5
5.8 x 10
8

Iood (37
o
C/48 hours) 300 7.1 x 10
2
300 4.5 x 10
2
1.3 x 10
7

Water activity (a
w
) oI Iood 0.988 0.984

MDU PHL

Fig A3.9: Challenge test - Pie
ChaIIenge Test - Pie
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
i
a
I

G
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

Fig 3.10: Challenge test - Quiche
ChaIIenge Test - Quiche
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Time (hours)
B
a
c
t
e
r
a
i
I

g
r
o
w
t
h

(
I
o
g

1
0
)
B cereus
S aureus
L m
Salmonella

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