cell by date bio-indicator for meat spoilage. introduction

Cell by DateCell by Date

Bio-Indicator for Meat Bio-Indicator for Meat SpoilageSpoilage

IntroductionIntroduction

Problem with Food Problem with Food IndustryIndustry

PROBLEMPROBLEM: Printed sell-by date is based : Printed sell-by date is based on predictive challenge testingon predictive challenge testing– Not reflective of product exposure to Not reflective of product exposure to

environmentenvironment– Printed Sell By Date is unreliablePrinted Sell By Date is unreliable

SOLUTIONSOLUTION: Monitor level of exposure : Monitor level of exposure and report it and report it – – Tell Consumer when meat is not fit for Tell Consumer when meat is not fit for

consumptionconsumption

Specification Design Modelling Implementation

Testing/Validation

MotivationsMotivations Fresh meat is a highly perishable foodFresh meat is a highly perishable food

Survey studies show temperature conditions >10°C Survey studies show temperature conditions >10°C are usual during transportation, retail storage, and are usual during transportation, retail storage, and consumer handling. consumer handling.

1/3 of Adults use spoilt food1/3 of Adults use spoilt food

Colouring and packaging methods are deceptiveColouring and packaging methods are deceptive

Win-win situation for producers-consumersWin-win situation for producers-consumers

Recycling of old projects – pH, temperature, BFPRecycling of old projects – pH, temperature, BFP


Testing/Validation

SpecificationSpecification1.1. Report When food is ‘off’Report When food is ‘off’

– Report cumulative Temperature exposure of foodReport cumulative Temperature exposure of food Temp is largest factor in spoilingTemp is largest factor in spoiling Easiest way to report if food is badEasiest way to report if food is bad

2.2. Easy To Use Easy To Use – Visual Reporter – Wow ! factorVisual Reporter – Wow ! factor

3.3. Improve Accuracy of other attempts:Improve Accuracy of other attempts:– SSM, Intelligent Cooling, AntimicrobialSSM, Intelligent Cooling, Antimicrobial

Only Threshold devicesOnly Threshold devices

4.4. System must be stable for length of shelf-lifeSystem must be stable for length of shelf-life5.5. No contact with food No contact with food

– – Need to obey HSA standardsNeed to obey HSA standards


Testing/Validation

1.1. Report Cumulative Temperature Report Cumulative Temperature exposure of foodexposure of food

Need memory to do this / Integrator fctNeed memory to do this / Integrator fct

2.2. Visual ReporterVisual Reporter Needs to have long degradation timeNeeds to have long degradation time

3.3. Improve AccuracyImprove Accuracy Limited Crosstalk with chassisLimited Crosstalk with chassis Easily tuneable thresholdEasily tuneable threshold

4.4. Stable for length of Shelf LifeStable for length of Shelf Life5.5. HSA standardsHSA standards

Project must not contaminate foodProject must not contaminate food

Considerations/Considerations/ProblemsProblems

SpecificationDesign Modelling Implementation

Testing/Validation

SolutionsSolutions

1.1. IntegratorIntegrator

3.3. Improved AccuracyImproved Accuracy

4.4. StabilityStability

5.5. HSA standardsHSA standards

6.6. Long Degradation TimeLong Degradation Time– Firefly FPFirefly FP


Testing/Validation

Cell Free System

Cell-free systemCell-free system– Widely-used Widely-used in vitroin vitro tool used to tool used to

study biological reactionsstudy biological reactions– Less of the complex interactions cf.Less of the complex interactions cf.

whole cell.whole cell.– Provides molecular machinery that Provides molecular machinery that

can be used in reactions with the can be used in reactions with the absence of many cellular absence of many cellular components.components.

SolutionsSolutions


Testing/Validation

SolutionsSolutions

1.1. IntegratorIntegrator

3.3. Improved AccuracyImproved Accuracy

4.4. StabilityStability

5.5. HSA standardsHSA standards

CFS increases stability due to lack of CFS increases stability due to lack of degrading enzymesdegrading enzymes

CFS stable for up to 4 daysCFS stable for up to 4 days CFS agrees with HSA standards CFS agrees with HSA standards


Testing/Validation

Cell Free System

DesignDesign


Testing/Validation

Photinus luciferin + OPhotinus luciferin + O22 + + ATPATP <=> oxidized Photinus <=> oxidized Photinus luciferin + COluciferin + CO22 + + AMPAMP + diphosphate + light + diphosphate + light

Pc

RBS TeTeLuciferase

Temp. Increase

Luciferin

O2

Cell-free extract medium

LIGHT!!!

ModellingModelling Concept of predictive microbiologyConcept of predictive microbiology

– Involves knowledge of microbial growth responses Involves knowledge of microbial growth responses to environmental factorsto environmental factors

– Expressed in quantitative terms using Expressed in quantitative terms using mathematical equationsmathematical equations

– Data used to interpret the effect of varying Data used to interpret the effect of varying conditions on microbial growth. conditions on microbial growth.

Integrative Temp function of productIntegrative Temp function of product– Leads to “intelligent” product management Leads to “intelligent” product management

systemssystems– Optimization of food quality and safety at the time Optimization of food quality and safety at the time

of consumptionof consumption


Testing/Validation

ImplementationImplementation


Testing/Validation

Bio indicator

Colour scheme

Simplest System : Phase 1

Good OK BAD

ImplementationImplementation


Testing/Validation

Simplest System : Phase 2

•Discrete Change In addition Phase 1

Good Bad

Eg. Discrete Change At 25oC for more than 2 hours

TestingTesting

Focus on proof of conceptFocus on proof of concept– Period of 24 hours as opposed to 4 Period of 24 hours as opposed to 4

daysdays– Regulated luciferase synthesis that Regulated luciferase synthesis that

corresponds to predicted outcomecorresponds to predicted outcome– Light intensity as a measurement Light intensity as a measurement

(compare with colour scheme strip)(compare with colour scheme strip)


Testing/Validation

Time lineTime line Week 3-4 – Modelling + GeneARTWeek 3-4 – Modelling + GeneART

Week 4-6 – Experiment Week 4-6 – Experiment implementationimplementation

Week 7-8 – Testing / ValidationWeek 7-8 – Testing / Validation

Week 9-10 – Polish the project + Week 9-10 – Polish the project + documentationdocumentation

ConclusionConclusion

Cell by date:Cell by date:

– Mimics printed ‘sell by date’ with Mimics printed ‘sell by date’ with additional temperature exposure trackingadditional temperature exposure tracking

– More accurate than conventional methodMore accurate than conventional method

– Will improve consumer healthWill improve consumer health

– Focuses on use of experimentation & proof Focuses on use of experimentation & proof of conceptof concept

EndEnd

Questions?Questions?

cell by date bio-indicator for meat spoilage. introduction

Documents

food need

temperature conditions

food industryproblem

monitor level of exposure

intelligent cooling

printed sell

consumer handling

effect of varying conditions