Rapid Microbiological Methods: Why and what!

Transcription

1 XIV. VEDECKÁ KONFERENCIA S MEDZINÁRODNOU ÚČASŤOU BEZPEČNOSŤ A KONTROLA POTRAVÍN March 2017 Piešťany, Slovenská Republika Rapid Microbiological Methods: Why and what! Dr. Lorenzo Castigliego, PhD OK SERVIS BioPro, s.r.o. Bořetická 2668/1, Praha

2 Content of the presentation 1 Why rapid microbiological methods (RMM) are often more convenient for food industry 2 What is on the market of effective and advantageous for pathogen detection and count of quality indicators

3 Traditional methods (standard) PROS & CONS Good Sensitivity Accuracy Reproducibility Labor demanding Long Multiple incubation and plating procedures Use of several different materials Direct costs Apparently less expensive Costs of labor Other indirect costs

4 What food industry and service labs need? Regulatory approved solutions Short time to result RAPIDITY Robust and simple methods SIMPLICITITY Solutions improving lab efficiency and decreasing expenses Direct costs Indirect costs

5 What food industry and service labs need? Regulatory approved solutions RMM Short time to result Robust and simple methods Rapid Microbiological Methods Solutions improving lab efficiency and decreasing expenses

6 Rapid Microbiological Methods: advantages Simplicity Rapidity

7 Simple methods HR management Less work for operators More or different tasks Better organization of time Activity peaks Temporary lack of personnel (stuff turnover; sick or maternity leaves)

8 Simple methods No specialized personnel Easier turnover of technicians No need for big or complex equipment Analysis directly in small facilities of the industry Cost of labor Costs of personnel Staff training needs

9 Short time to result Increase of sample throughput Earlier in-process notification Speed of corrective actions (decisions) Raw material orientation Finished or semi-finished products Reactivity after cleaning Product loss Delay in production and delivery

10 Short time to result Release time of products Production Lab analyses

11 Short time to result Release time of products Hidden costs Faster placing on the market of the products Lower costs of storage (and energy) More rapid cash flow More time for operators

12 Cost effectiveness inventory costs Carry Costs (holding costs of inventory w/o sales (invoicing)) Finance (daily cost of capital: 5%/365=0,0137% per day) Damage (longer to hold => higher risk of damage) Obsolescence (shortened shelf life) Warehousing costs Electricity, rent, overtime, counting, cooling, Lost sales (because you can t deliver) Lost sales = lost growth profit Expedite fees (supplier, contract lab,..) Extra costs (overtime in lab, extra services) Additional manufacturing costs Additional packaging, additional QC time(!) ->$ Freight out (express delivery!)

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14 One real practical example: Company: Very important brand in the global food market Kind of products: various (perishable and long-term storable) Cost for 58 days of stock cover Stock value (in the period of storage) Cost for 1 day of stock cover Spaces Labor Energy Invoicing time Cash flow Etc.. How much money could be saved if the period of storage could be reduced of about 1\3 or 2\3? 2,7% Of the stock value 5,3% Real profit Much higher % Much higher %

15 Actual costs: a final comparison Actual costs Indirect costs Analysis Instrumentation Labor Storing Spaces Energy Management RMM Investment ** * Standard methods **/*** ** * ** * *** Finance * ***! Time to solve crisis events * ***!

16 A straightforward equation: Simple methods Fast methods = = Less money to spend Less money to spend

17 Which rapid microbiological methods

18 What possibilities for RMM in pathogen detection? Rapid chromogenic media Nowadays very evoluted (specific growth factors + chromogenic substances) Fast Automated immunoenzymatic methods Direct detection of pathogens w/o plating Fast Very fast PCR Detection of pathogens` DNA w/o plating Very fast

19 Which possibilities for RMM in quality indicator counting? Rapid chromogenic media Nowadays very evolved (specific growth factors + chromogenic substances) Fast MPN-based methods Automatic detection of fluorescence associated to microbial growth Fast Flow cytometry Detection of single microbial cells ULTRA fast

20 Which possibilities for RMM? Rapid chromogenic media Validation Immunoenzymatic PCR MPN-based methods National and International ISO AOAC OMA They can replace standard reference methods Flow cytometry Assure a high level of performance Simplify workflow of exporting companies

21 Pathogen detection Automatic Enzyme Linked Fluorescence Assay Peace of mind - VIDAS Ready to use reagents (minimum risk of error) in 1 single strip Easy to use system and protocols

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23 As easy as 1, 2, Load sample 2- Insert test 3- Press button and. Walk Away

24 VIDAS/standard methods workflow Standard methods VIDAS 48 h VIDAS NEXT day Pre-enrichment Enrichment Plating Results

25 VIDAS applications (even less than 24h) LDUO (Listeria spp + L. mono) LMO2 (Listeria mono.) SLM (Salmonella) CAM (Campylobacter) - 24 h LMX (L. mono. ) LPT (L. spp ) SPT (Salmonella) ECPT(E. coli O:157 incl. H7) 48 hours solutions Next day solutions

26 Count of quality indicators Automated solution based on traditional method (MPN) to enumerate Quality Indicators in food and environmental samples TEMPO Filler TEMPO Reader Only 1 dilution No dilution No inoculation No counting

27 TEMPO: the principle of MPN 1 ml 1 ml 1 ml Incubation X CFU/ g Interpretation using statistical tables Counting

28 TEMPO: the principle The core of the system: an ENUMERATION card 48 tubes (or wells) of 3 different sizes for the 3 dilutions

29 TEMPO: the principle The core of the system: an ENUMERATION card integrated in a molded plastic card

30 TEMPO: the principle The core of the system: an ENUMERATION card Selective medium + Precursor of a fluorescent molecule after incubation, fluorescence reading (under UV lamp)

31 Time and labor saving 6 cards filled in only 3 minutes with 1 TEMPO Filler During this period, preparation of 6 additional cards More than 100 tests performed in 1 hour only

32 TEMPO applications Parameter Incubation time AC Aerobic count Next day EB Enterobacteriaceae Next day EC Escherichia coli Next day STA Staph. Coag. + Next day TC Total coliforms Next day CC Coliforms` count Next day YM Yeasts and molds h VS 5 days! LAB Lactic Acid Bacteria h BC Bacillus cereus Next day

33 ULTRA-rapid microbiological tests Flow cytometry LASER DETECTION OF SINGLE VIABLE MICROBIAL CELLS Presence/absence Direct count Up to 50 samples per hour Ready-to-use reagents Complete automation Digital data process with full traceability

34 The technology of D-count Criteria of Viability : Membrane integrity Enzyme activity micro-organism Enzyme Viability substrate free fluorochrome

35 The technology of D-count Labelled cells are forced through a micro linear fluid stream Microorganism cells are hit by a laser beam Emitted fluorescence is detected by sensitive photo-detectors Microorganism cells are counted one by one Counting every single viable cell Laser Waste Sample Detector 1 Detector 2

36 D-count Validation The first and only alternative method based on Flow cytometry being ISO AFNOR validated Best analytical sensitivity Patented cell labeling technology +++ sensitivity From 1 log to 6 log more sensitive than the other alternative methods Shorter pre-incubation time for analysis

37 Applications in DAIRY PRODUCTS PRESENCE/ABSENCE Yeasts in fermented milk products Enterobacteriaceae Yeasts & molds in fruit preparations Sterility testing UHT milk Sterility testing dessert DIRECT COUNT Direct count in process water Direct count in milk

38 Applications in BEVERAGES PRESENCE/ABSENCE Yeasts & molds in soft beverages and fruit juices Commercial sterility DIRECT COUNT Direct count in process water Yeasts in fruit concentrates Bacteria in fruit concentrates

39 Economical BENEFITS Global benefits for the company

40 TAKE HOME MESSAGE Rapid methods in microbiology To save money Labor Financial Warehousing Inventory To make money

41 What we propose Pathogen detection Count of quality indicators

42 What we propose Ultra-rapid microbiology Presence/absence Direct count

43 You are welcome to speak with us about your needs and projects