MICROBIAL RISK ANALYSIS & MITIGATION IN

BEER-MIX BEVERAGES

Tadhg O’Sullivan Inge Suiker EBC Vienna September 2014

OVERVIEW

1. BACKGROUND

2. EXPANDING PORTFOLIO

3. METHODOLOGY

4. BEER MIX BEVERAGES

5. TO CONSIDER

6. WORK IN PROGRESS...

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1. BACKGROUND - concern

♦ New and increased portfolio of beer and non-beer products

♦ The characteristics of these new products are more complex and different to regular beers

♦ Necessary to map these differences with respect to:

■ Understanding microbiological risks

■ Adapt installations to safely produce these beverages

■ Ensure product stability is maintained

■ Ensure correct production and packaging methods are in place

■ Ensure correct microbiological quality control standards

■ Challenge existing standards

♦ Spoilage versus safety

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1. BACKGROUND - concern

♦ Microbiological stability of a product

■ Indicator of quality

■ Influences company reputation

♦ Microbial spoilage of products is unfavorable

■ Odours, off-flavors and turbidity

■ Induce safety and integrity risks in case of contaminating yeast

● Increased alcohol -> Product integrity risk (non/low-alcoholic beverages)

● Increased CO2 pressures -> Safety risk (bursting bottles)

■ Financial loss and brand damage from recalls and withdraws

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1. BACKGROUND-Microbial stability of beverages

♦Factors that influence microbial stability in beverages

pH/acidity

Alcohol

Bitterness Sugars

(Nutrients)

Preservatives

Particles

(Yeast/pulp)

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Different charecteristics Different growth conditions Different microbiological risks

1. BACKGROUND – Microbial stability of beverages

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•Disrupts cell membrane and enzyme systems

•Reduces nutrient uptake Low pH/ high acidity

•Inhibits cell membrane functions

•Induces cell membrane leakage High ethanol

•Cell membrane leakage

•Inhibits active cross membrane transport Bitterness (Hop)

•Inhibits aerobic microbes

•Lowers pH

•Affects carboxylation and decarboxylation reactions

Dissolved gases (CO2)

•Low nutrients inhibit growth

•Extract of fruits and spices with antimicrobial activity Nutrients and functional ingredients

•Sorbates target yeast and moulds

•Sulphites target bacteria Chemical preservatives

•Heat treatments

•Flash pastuerization

•Tunnel pastuerization

Physical processing

1. BACKGROUND-Microbial stability of beverages

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•Disrupts cell membrane and enzyme systems

•Reduces nutrient uptake Low pH / high acidity

•Inhibits cell membrane functions

•Induces cell membrane leakage High Ethanol

•Cell membrane leakage

•Inhibits active cross membrane transport Bitterness (Hop)

•Inhibits aerobic microbes

•Lowers pH

•Affects carboxylation and decarboxylation reactions

Dissolved gases (CO2)

•Low nutrients inhibit growth

•Spices and extracts with antimicrobial activity Nutrients and functional ingredients

•Sorbates Benzoates mostly target yeast and moulds

•Sulphites target bacteria Chemical preservatives

•Heat treatments

•Flash pastuerization

•Tunnel pastuerization

Physical processing

2. EXPANDING PORTFOLIO

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Product Characteristics Risk Types

Alcohol

ABV pH

Residual

Ferment. Sugar

g/100ml

LAB Yeast GNAs Safety (Food &

Physical) Potential Risk

Beer (lager, ale, stout, despé) >3,5 >4,0 < 2,5 Yes Yes Yes Yes (CO2) Exploding glass bottles

Beer (lager, ale, stout, despé) >3,5 < 4,0 > 2,5 No Yes No Yes (CO2) Exploding glass bottles

Beer (light) 1,2-3,5 >4,0 < 2,5 Yes Yes No Yes (CO2)

Exploding glass bottles

Alcohol increase

Beer (light) 1,2-3,5 >4,0 > 2,5 Yes Yes No Yes Exploding glass bottles

Alcohol increase

Beer (Low Alc.) 0-1,2 >4,0 > 2,5 Yes Yes No Yes Exploding glass bottles

Alcohol increase

Wheat beer (std & flav) >3,5 >4,0 < 2,5 Yes Yes Yes Yes (CO2)

Exploding bottles due to

CO2

Beermix (Radler/Shandies) 1,2 -3,0 <4,0 > 2,5 Yes Yes No Yes Exploding glass bottles

Beermix (Radler/Shandies) 1,2-3,0 <4,0 < 2,5 Yes Yes No Yes (CO2) Exploding glass bottles

Beermix 0 (Radler Low Alc.) 0-1,2 <4,0 > 2,5 Yes Yes No Yes Exploding glass bottles

Alcohol increase

Malt beverages 0 >4,0 > 2,5 Yes Yes Yes Yes (C02/ Food?) Exploding glass bottles

Pathogen risk?

Malt beverages 0 <4,0 > 2,5 Yes Yes No Yes (CO2) Exploding glass bottles

Alcohol increase

Cider >3,5 <4,0 < 2,5 No Yes No Yes (CO2)

Exploding glass bottles

Cider >3,5 <4,0 > 2,5 No Yes No Yes (CO2) Exploding glass bottles

Cider (Low) <3,5 >3,5 > 2,5 Yes Yes No Yes (CO2) Exploding glass bottles

Alcohol increase

RISK ANALYSIS

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Assessing the ability of a range of potential

spoilage microbes for:

survival, growth and spoilage potential

Evaluate possible integrity / safety risks from

[CO2] / [alcohol] that can increase due to spoilage

Sort the different beverages into risk categories

based on their microbial stability and safety risks

3. METHODOLOGY-Challenge tests and CO2 pressure test

Acclimatize

microbe

•Inoculate in

broth:beer mixtures

Inoculate into

beverage

•1000

cells/ml

Incubate at

30oC •6 weeks

Weekly

checks

•Visual (Turbidity and

gas)

•Flow cytometry (cell

count)

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Bursting septa

EXAMPLES

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1 bar 7 bar 10 bar

4. BEER MIX BEVERAGES

■ Catagorise beverages into groups?

● Sugar

● Alcohol

● Natural inhibitants

■ Assess microbial spoilage risks of the different beverage groups

■ Risks

● Safety?

● Integrity?

● Spoilage (off flavors)?

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4. BMB– Risks and implications

■ Low risk spoilage due to Lactic Acid Bacteria or Gram Negative Anaerobes

● Some LABs to cause slight spoilage even at low pH (3.2)

■ Perform flavor tests to determine effect of spoilage on product

● Possibly include detection of LABs quality standards even for low pH products

■ High risk of spoilage by yeast.

● High CO2 pressures and increased alcohol concentrations observed

■ Flash pasteurization not recommended:

● secondary infection risks

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5. TO CONSIDER

♦ Challenge tests

■ Possible false positive / negative results

■ Critical parameters

● Oxygen

■ Test tubes versus PET-bottle set-up

● Physiological state of micro organism

■ Not adapted ~70% of inoculum DOA

♦ Quality control method

■ High levels of natural inhibition

● Low contamination levels hard to detect

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5. TO CONSIDER

♦ PU requirements

■ Current standards sufficient?

● Time / temperature

● Particles

■ Flash versus tunnel

♦ Preservation

■ Sorbate / Benzoate / Natural - Clean label

● Levels allowed

● Do they work?

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6. WORK IN PROGRESS

♦ Flash pasteurisation not recommended

■ Loss of flexibility with packaging

■ Reduction in sustainable practices

● High energy consumption tunnel pasteurization

● Transport of heavy and bulky glass bottles in stead of lightweight tetrapaks and PET bottles

♦ Road tankering

■ Logistics / Standards

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6. WORK IN PROGRESS

♦ Catagorise

♦ Challenge tests

♦ Natural inhibitors

♦ Pasteurization trials to obtain the right time-temperature treatments for different beverage groups

♦ Essential

■ Improve hygiene standards and GMPs => lower the risk of (secondary) contamination

■ Improve quality control standards => Correct detection media, Correct sampling plans

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Thanks to

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Heineken Microbiology Team:

Tadhg O’Sullivan

Petra Zeegers

Vincent de Groot

Interns

Kavitha Panicker

Sophia Bernasconi