safety for beverage production -...

Safety for beverage productionDetection of microorganisms in beverages and at beverage production sites

Microbiological quality control at its highest level.An all-in-one package for effective microbiological monitoring. From one source.

Detection kits | Services | Consulting

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Microbiology in production processes

Conventional microbiology and its modern concepts are not up for today‘s tasks. New approaches are needed.

A highly functional, efficient microbiological monitoring system has to be able to control the microorganisms rapidly, specifically and with high sensitivity.

Moreover, following samples and problems must be analyzed and solved:

the microbiology of input and raw materials

microbiological in-process controls

end product controls and release decisions

individual microbiological tasks

unexpected events influencing the microbiology

newly developed products and thus microbiological changes

Though, the requirements to microbiological control systems are clearly defined as:

rapid and reliable control during all steps of production in order to enable short response times

highly specific identification of microorganisms; reliable differentiation between unwanted germs and harmless accompanying bacterial flora

precise detection of contamination sources

analysis of contamination streams and sustainable elimination of their source

The reality is, however, that conventional microbiology and its indirect approach is outdated despite its punctual improvements. Efficient microbiological monitoring requires a holistic approach covering the complete production processes and delivering full insight into samples.

The indirect detection methods are inadequate for microbiolo-gical quality control that meets the latest requirements. Most methods are limited in their efficiency as they are based on a punctual approach and often error-prone.

The vermicon detection system for microorganisms comes right into play where other methods reach their limits. It represents the only system which follows a direct approach and is based on the innovative VIT® gene probe technology developed by vermicon. It enables the detection of living microorganisms in all steps of beverage production.

The direct approach

The advantages of the VIT® gene probe technology

Viability testing – Only living bacteria are detected.

Rapidity – Results possible within a few hours.

Reliability – Correctness of results many times shown.

Specificity – Adjustable from species level to all viable bacteria.

Robustness – Insensitive to different sample material. No enzymes used.

Established – Longtime routine-application of the technology.

Visualization – Direct insight into the sample.

Extensive – A broad spectrum of bacteria relevant to beverage production is covered by VIT® analysis.

Find out more about the technology on page 11.

culativation

PCR and other indirect detection methods

Assay

VIT® gene probe technology

Indirect approach – examples Direct approach – VIT® gene probe technology: direct, unfalsified insight into samples. This enables actual real-time monitoring and profiling.

Raw materials Production processes End products Additive processes

In principal, a microbiological quality control system should literally be able to “control” microorganisms and their occur-rence. However, it is the microbiology which actually controls the production process and not the other way around. With this regard and considering the increased quality level of pro-duction requirements functional control systems have to meet various tasks:

complex production processes

high production speed

high hygienic standards

high quality standards expected by consumers

„Go Green“ legal regulations and awareness

quickly adapting microorganisms

Quality standards for industrial microbiology Microbiological control during all production steps

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No compromise in microbiological quality control

Since 1997 vermicon AG from Munich provides solutions for microbiology – innovative and future-oriented. Our customers profit from our expertise and our will to break new ground together.

We offer products, services and consulting – the one-stop solution.

In reference to our company principles we promise:

… to provide unique and efficient solutions for microbiology...

… to meet successfully microbiological challenges which no one has solved before.

vermicon AG: knowledge – experience – solutions

5

For a holistic insight: the vermicon AG servicesEfficient microbiological monitoring in the beverage production sector requires observation of the entire production process. Only by having a complete overview on the whole microbio-logical situation potential changes can be recognized and analyzed.

Only then, on time reaction and prevention of production process disruptions is ensured.

Whether products, services or consulting – use the strengths of VIT® to your own advantage!

VIT® is used everywhere in beverage industry.

Future-oriented: 4D-Microbiology

The focus of microbiological quality control has to lie on rapidity, specificity, robustness and on the reliability of the applied system. The provided results must enable safe release decisi-ons. Only with a functional efficient control system hazardous microorganisms have no chance. Moreover, only beverages with a steady high quality will succeed on the market.

The solution for unspoiled high quality products: VIT® (VIT®= vermicon identification technology).

The VIT® technology is applied for the vermicon‘s detection kits, services and consulting – depending on the individual needs of the applicant. The use of VIT® guarantees the highest safety level of microbiological quality control for beverage production. The innovative technology is in routine use approved and deli-vers new, ground-breaking insight!

VIT® is also used for the established vermicon 4D-Microbiology and thus as part of all the company‘s services.

4D-Microbiology lies its focus on all of the 4 dimensions of microbiology: Identification, quantification, temporal and spatial analysis. Only then a holistic picture of the microbio-logical environment during a live process can be drawn.

identification quantification

spatial analysis

temporal analysis

gives insight

creates outlook

unique, holistic

provides a competitive advantage by ensuring high product quality standards

builds confidence

creates long-term efficiency of your production

individual solutions

patented, highly innovative technologies

application of the entire microbiological competence

highly professional, goal-ori-ented processing

close cooperation with our clients

The advantages of the 4D concept:

vermicon offers for your company:

Established quality The products and services are based on the reliable and innovative VIT® gene probe technology – various high-class studies regarding our products developed for the beverage sector are available. Request publications of your interest on www.vermicon.com A. I. J. N.: Meet standards with VIT®

This guide acts as reference which helps minimizing product spoilage caused by Alicyclobacilli. GMP certified services

We only apply the highest standards to our own work. Thus, our company has been GMP (Good Manufacturing Practice) certi-fied.

Protected by vermicon!

Products

- for daily routine checks

Consulting

- for problem solving in the caseof process malfunctions

Services

- for specific questions

VIT-Profiling

Holistic concepts for microbiological monitoring

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VIT®-Kits: Almost limitless possibilities of application

The detection of microorganisms with the vermicon kits is quick, sensitive and reliable. Only living microorganisms are detected. Results are provided in short time. The difference to conven-tional technologies is obvious:

Time-to-result:

Conventional cultivation: up to 10 days

VIT®: 3 hours

VIT® combined with necessary pre-enrichment: 1–2 days.

The fields of application and sample materials are diverse:

raw materials

all interim steps of production

final products

High-End-Technology in a standardized format – learn more about the vermicon VIT®-Kits!

Analyze the samples taken from the various steps of production directly at your own lab – with only little equipment required. All essential expendable materials are contained in the kit.

The application and evaluation of the kits is simple, the results allow for quick release decisions.

For a large number of microorganisms we already offer specific products. Additionally, we develop new detection kits in close cooperation with our clients which fit their individual needs.

Inquire about individual kits for your application.

The “basic catalog” for the beverage sector comprises:

VIT-Alicyclobacillus, #: 01210013

VIT-Bier Megasphaera/Pectinatus, #: 01210007

VIT-Bier plus L. brevis, #: 01210006

VIT-Lactococcus lactis, #: 01210027

VIT-Leuconostoc/Lactococcus lactis, #: 01210028

VIT-Lactic acid bacteria, #: 01210018

VIT-Spoilage yeasts, #: 01210016

VIT-Pseudomonas aeruginosa, #: 01210001

The VIT®-System

The components

Analysis, visualization, documentation, evaluation, image processing and archiving of microbiological analysis: The VIT®-System provides highly specific, innovative and future-oriented analysis of microorganisms for all sectors of industrial micro-biology.

Aligned components and modular technologies with intuitive handling and application-oriented functionality – developed and matched to the lab routine of microbiological quality con-trol.

VIT®-Kits: Rapid and specific detection of microorganisms. Easy to handle – combined in vermicon‘s VIT®-Kits. The reliable and robust detection kits stand for modern microbiology that meets all of the industry‘s requirements!

VIT®-Microscope: The VIT®-Microscope combines high quality and simple handling – specifically aligned to the VIT®-Kits. „Made in Germany“ guarantees highest standards regarding its inside and its outside.

VIT®-Cam: The camera for documentation of results offers high speed with best quality. The VIT®-Cam is a powerful tool with best price-performance ratio.

VAS – vermicon analysis software: the ideal completion with the VIT®-Cam. The software streams the microscopic images extraordinaly fast to your monitor. The image resolution and the easy, application-oriented features satisfy all of your needs.

Products

The vermicon VIT®-Kits: Innovative technology in a standardized format. All necessary expendable materials are included.

Simple handling, quick, highly specific, high sensitivity

All-in-one system solutionHighest SpecificityEasy Handling

Systems for the detection of microorganismsInnovative Technology – brought directly to your company

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Services

Individual customer service and high flexibility

Our innovative technology portfolio allows for unique results. Thereby, it is of particular importance to us to guide our clients through their “microbiological journey”. It is only this close cooperation with our clients which enables us to identify the technolo-gies capable of answering the questions they ask. Only this way we can successfully and quickly solve their tasks.

We analyze a large number of diverse sample materials (raw materials, final products, process samples) for you. With our innovative technology portfolio we are able to even find quick and reliable solutions for rather unusual or unique tasks.

Our services are individually matched to your specific needs. Profit from our long-term market experience and microbiological know-how. Simply send your samples to us and our expert team will analyze them. Quick, professional and discrete – for reliable and unique results.

Our service portfolio offers you many possibilities.

Some examples:

Rapid quantification of flock forming bacteria in beverages

Rapid quantification of Alicyclobacillus

Rapid quantification of acetic acid bacteria

Rapid detection of beverage spoilage lactic acid bacteria

Rapid detection of beer spoilage bacteria

Rapid detection of fermentative yeasts

Rapid detection of E. coli and coliform bacteria

Individuality is our excellence. You want to detect a specific type of bacterium? You have unusual sample material? Your task cannot be managed by applying conventional methods? No problem. Name your task and we will develop a custom-made solution concept for you.

Consulting

During the beverage production steady quality control aims to prevent production disruptions or in case of their occurrence eliminate them quickly and effectively and to ensure impec-cable end product quality. However, often the microbiological tasks are complex and can only be solved by applying a holistic approach.

Such tasks require unconventional approaches.

Just on this, vermicon has specialized by introducing its new microbiological consulting. With almost two decades of active market experience and its competent and highly moti-vated team vermicon is proud to be able to offer – verifiable

– new and highly efficient solutions. Our innovative technology portfolio helps answering questions unsolvable by conventio-nal methods.

Not limited by conventional restriction our team gains insight into microbiological worlds not accessible by others.

In close cooperation with our clients we analyze the technical processes and can thus further deliver a holistic image based on our microbiological findings which is the basis of the project‘s success. Our findings are translated into specific recommended actions which our clients can apply to execute their task.

Gain holistic insight into your production and fight and eliminate microbiologically induced process disruptions.

Microbiological Consulting – Consulting for microbiological tasks. VIT®-Profiling – Detection of contamination streams.

Analysis – of even only hardly or entirely uncultivable microorganisms.

Development – of custom-made solutions.

The vermicon services:

The vermicon experts take a look directly inside your sample. Profit from our expertise!

Microbiological Consulting by vermicon:

Solution-oriented and holistic approaches to tasks. We are your competent partner for so far unsolved questions, too!

Quick, professional, extra-ordinary Individual, goal-oriented, holistic

Innovative technology – unique results Innovative technology – holistic solutions

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VIT®-Profiling: practical application during beverage production

Often uncertainty remains: the cause of unwanted production related incidents stays unclear. The catalysts cannot be identified. The aim: Identification and elimination of the cause.

By the practical-oriented VIT®-Profiling based on our 4D-Microbiology microbiologically induced process disruptions can quickly and permanently be eliminated.

Our experienced VIT®-Profiling team analyzes your plant and its process streams in cooperation with your technical specia-lists. Critical points are localized and samples are taken which are then analyzed in our cutting-edge laboratories by means of our highly effective technologies. Hereby, the VIT® gene probe technology allows for profiling of all contained bacteria directly within the samples.

The subsequent data base supported evaluation enables the profiling of microbial contamination streams and thus the eli-mination of the disruption. But we can do even more: Based on the created profiles we develop individual early warning and detection systems which we then establish at your production site.

This world wide unique VIT®-Profiling allows for unique and holistic insight into the microbiology of production processes and hence for highly effective solutions.

Test us!

The VIT®-Profiling is applied by many beverage producers of all sizes, such as:

optimization of production processes by continuous in-process-monitoring

detection of contamination during the different production steps and practical support for the elimination of contamination sources

detection and identification of so far unknown floc forming microorganisms in already bottled beverages

Easy to use, reliable:

The patent protected VIT® gene probe technology (VIT®: vermicon identification technology) is a highly efficient detection system for microorganisms. The method is robust and reliable. By the application of so-called VIT® gene probes bacteria, yeasts and moulds can be marked with high specifici-ty and identified subsequently. Whether or not the organisms can be cultivated makes as well as the matrix in which they are present no difference.

During the various steps of production – from raw material to in-process samples and to the final product – microorganisms can be detected and identified. As both testkit and as service – VIT® meets the highest standards.

Years of experience in the sector of bacteria detection in be-verage industry makes us your ideal partner in this area. Particularly, in the most sensitive field of beverage production our services pay off – we are able to give individual support to all of our clients. Solution and goal-oriented cooperation is our primary objective. You are provided with exactly what you need: Whether it concerns instructing you to the application of our kits or conducting analysis as services or individual consulting.

Satisfied customers from all over the world rely on our solutions and believe in their cooperation with us for many years now.

Our testing systems are many times validated and tested in national as well as international studies. Find out yourself and request the respective publications at www.vermicon.com.

We look forward to a successful cooperation!

The VIT® gene probe technology

Don‘t compromise over quality and get in touch with us!

The reliable partner for beverage producers

1. Gene probes are applied to the sample.

3. Specific gene probes bind only within living target microorganisms.

2. Gene probes penetrate all microor-ganisms.

4. Unbound gene probes are washed off.

5. The target micro-organisms begin to shine.

Raw materials processing/finishing end product

retracing 2nd contamination

retracing 1st contamination

VIT®-Profiling: contamination source detection | elimination | prevention

Conventional methods VIT®-Profiling

most samples contain only 1 % cultivable bacteria

detects up to 100 % of all bacteria

indirect approachdetection by VIT® gene probe technolo-gy in situ, directly within the sample

1-dimensional analysisspatial and temporal analysis possible (4D-Microbiology)

inflexible, limited approachholistic, individual microbiological solutions

undirected, time-consuming analysis methods

goal-oriented, fast results

Our competenceMicrobiological Consulting by vermicon

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Emmy-Noether-Str. 2 80992 Munich Germany

T: +49 89 1 58 82-0 F: +49 89 1 58 82-100

[email protected] www.vermicon.com

Let‘s keep in contact

Visit us directly: www.vermicon.comGet informed at our internet page about the continuously updated range of products. Just ask for more detailed information regarding our products and services or request our complete catalog.

Moreover, you can find here up-to-date articles concerning all about microbiology as well as studies and publications.

Visit us and dive into the world of microbiology – the world of vermicon!

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Be surprised – gain new insight in the world of microorganisms

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Products

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Unspoiled pleasureThe detection of spoiling microorganisms in beer and beer-based drinks

Microbiological quality control on its highest level.

Detection kits, services, consulting: the full package for effective microbiological monitoring. All from one source.

Our competence

The patent protected VIT® gene probe technology is a highly efficient detection system for microorganisms. The method is robust and reliable. By the application of so-called VIT® gene probes viable bacteria, yeasts and moulds are marked with high specificity and identified subsequently. Whether or not the organisms can be cultivated makes as well as the environment in which they are present no difference.

During the various steps of production – from raw material to in-process samples and to the final product – microorganisms can be detected and analysed directly. As both testkit and as service – VIT® meets the highest standards.

The VIT® gene probe technology – simple handling, safe results








vermicon AG


T: +49 89 1 58 82-0 F: +49 89 1 58 82-100


Since 1997 a reliable partner for breweriesYears of practical experience of bacteria detection in the brewing industry make us your ideal partner. We developed our testing systems for the beer sector (beer spoilage bacteria, spoilage yeasts) in close cooperation with the renowned Chair of Brewing and Beverage Technology (Technische Universität München, Weihenstephan, Prof. Back). Satisfied customers from all over the world rely on our technology.

Studies prove: the VIT® gene probe technology is an ideal tool for breweries! Find out yourself and request publications of your interest from us, e. g.:

„Specific rapid detection test for beer-spoilage bacteria using fluorescently labelled gene probes“by: K. Thelen, C. Beimfohr, I. Bohak und W. Back, Freising-Weihenstephan, published in: Brauwelt International, no. 3 (2002).

„Evaluation Study of the Frequency of Different Beer-Spoiling Bacteria using the VIT Analysis“, by: K. Thelen, C. Beimfohr, J. Snaidr, published in: MBAA TQ, vol. 43, no. 1 (2006).

„Description of Lactobacillus backi sp. nov., an Obligate Beer-Spoiling Bacterium“, by: I. Bohak, K. Thelen, C. Beimfohr, published in: Monatsschrift für die Brauwissenschaft, March/April 2006.

www.vermicon.com,

Support Publications

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The focus of modern microbiological quality control lies on rapidity, specificity, robustness and reliability of the applied system. The provided results have to enable safe release decisions. Only a functional effective monitoring system is able to keep hazardous microorganisms in control.

The solution for high quality products: VIT® (VIT®= vermicon identification technology). The VIT® technology is used in detection kits, for our services and our consulting – depending on your individual needs! By the application of VIT® microbiological quality control in beer production is brought to its highest safety level. The innovative technology is for long years approved in routine use and offers new, ground breaking insight!

Control every step of the productionThe vermicon system for the detection of bacteria, yeasts and molds can be applied to every step of the production process. You can analyze a diverse range of sample materials – the scope of application of VIT ® has almost no limits.

vermicon offers a highly functional stage control which leaves room for action!

Don‘t compromise over microbiological quality control

High standards for maximum quality

The vermicon detection kits for beer and beer-based drinks

Only living microorganisms, fast, high sensitivity, simple handling

filtration

Analyze your beer samples from the various steps of production – with only little equipment required! The conduction and evalu-ation requires almost no coaching.

All necessary expendable materials are included in the kit (package size: 25 analyses):

VIT-Profiling

Detection kits Services Consulting

The complete VIT®-System:

The VIT®-System enables the highly specific, innovative and future-proof analysis of microorganisms in all sectors of industrial micro-biology. Consisting of VIT®-Kits, VIT®-Microscope, VIT®-Cam and VAS (vermicon analysis software) it enables analysis, visualization, docu-mentation, image processing and archiving of microbiological results. Get to know the VIT®-System now!

VIT-Bier plus L. brevis, # 01210006

parallel detection of beer spoilage lactic acid bacteria and identification of Lactobacillus brevis

saves between 4 to 5 days compared to conventional methods

all relevant steps of the brewing process can be analyzed by VIT-Bier plus L. brevis

VIT-Bier Megasphaera/Pectinatus, # 01210007

parallel detection of Pectinatus spp. and Megasphaera cerevisiae

saves between 5 to 8 days compared to conventional methods

an ideal addition to VIT-Bier plus L. brevis

VIT-spoilage yeasts, # 01210016

detection and differentiation of obligate and potentially fermentative yeasts

enables to verify the absence of living fermentative yeasts in beer and beer-based drinks

phase contrast picture with VIT®: beer spoilage lactic acid bacteria and in parallel: Lactobacillus brevis

Unequivocal evaluation:

specificity

rapidity

only living microorganisms

reliability

robustness

visualization

one-stop solution

VIT® makes the difference! The advantages:

Quality assurance with vermicon:

One-stop solution.

With the VIT®-Microscope bacteria specifically are made visible.

Each of the 3 images shows the identical microscopic picture.

VIT-Leuconostoc/Lactococcus lactis, # 01210028

specific identification of Leuconostoc spp. and of Lactococcus lactis

usable as early warning system for micro-biological contamination with beer spoilage bacteria

an ideal addition to VIT-Bier plus L. brevis and VIT-Bier Megasphaera/Pectinatus

MIC

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Unspoiled pleasureThe detection of spoiling microorganisms in beer and beer-based drinks

Microbiological quality control on its highest level.

Detection kits, services, consulting: the full package for effective microbiological monitoring. All from one source.

Our competence

The patent protected VIT® gene probe technology is a highly efficient detection system for microorganisms. The method is robust and reliable. By the application of so-called VIT® gene probes viable bacteria, yeasts and moulds are marked with high specificity and identified subsequently. Whether or not the organisms can be cultivated makes as well as the environment in which they are present no difference.

During the various steps of production – from raw material to in-process samples and to the final product – microorganisms can be detected and analysed directly. As both testkit and as service – VIT® meets the highest standards.

The VIT® gene probe technology – simple handling, safe results








vermicon AG


T: +49 89 1 58 82-0 F: +49 89 1 58 82-100


Since 1997 a reliable partner for breweriesYears of practical experience of bacteria detection in the brewing industry make us your ideal partner. We developed our testing systems for the beer sector (beer spoilage bacteria, spoilage yeasts) in close cooperation with the renowned Chair of Brewing and Beverage Technology (Technische Universität München, Weihenstephan, Prof. Back). Satisfied customers from all over the world rely on our technology.

Studies prove: the VIT® gene probe technology is an ideal tool for breweries! Find out yourself and request publications of your interest from us, e. g.:

„Specific rapid detection test for beer-spoilage bacteria using fluorescently labelled gene probes“by: K. Thelen, C. Beimfohr, I. Bohak und W. Back, Freising-Weihenstephan, published in: Brauwelt International, no. 3 (2002).

„Evaluation Study of the Frequency of Different Beer-Spoiling Bacteria using the VIT Analysis“, by: K. Thelen, C. Beimfohr, J. Snaidr, published in: MBAA TQ, vol. 43, no. 1 (2006).

„Description of Lactobacillus backi sp. nov., an Obligate Beer-Spoiling Bacterium“, by: I. Bohak, K. Thelen, C. Beimfohr, published in: Monatsschrift für die Brauwissenschaft, March/April 2006.

www.vermicon.com,

Support Publications

MIC

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JuanRamon

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18 · BBII 2/2006

MICROBIOLOGICAL QUALITY CONTROL

Reliable analysis of E. coli, coliform bacteria and fermentative yeasts

The analysis of E.coli and coliforms as well as fermentative yeasts

is an integral part of the microbiological quality control in the

production of beverages. The aim of the quality control is to quickly

and reliably confirm the hygienic purity of the products and to confirm

that there is no risk of product spoilage due to the presence of

microorganisms. The rapid detection system VIT was integrated

into the microbiological quality control process at Adelholzener

Alpenquellen GmbH, Germany, in order to achieve this aim.

The fast and unequivocal results delivered by the system contribute

significantly to the daily product release decision process.

Fig. 1: Schematic representation of the analysis of E.coli and coliform bacteria in mineral water as per German regulations in the quality control process at Adelholzener Alpenquellen.

Franz Stettner

Project engineer for microbiology and technology,AdelholzenerAlpenquellen.

AlbertReischenboeck

Quality AssuranceManager,AdelholzenerAlpenquellen.

The history of the Adelholzenerhealing spring can be traced

back to the year 300 A.D. Sincethen, the spring has been used to provide people with preciouswater. Today, AdelholzenerAlpenquellen GmbH is one of the most modern and ecologicallyactive beverage manufacturers in Europe. The company producesa wide range of beverages includingmineral water, carbonated softdrinks, fruit juices, tea and wellnessdrinks. The yearly output is currentlyapproximately 400 million fillings.

Highest quality is the omnipresentcriterion for the production of thesebeverages. The wide range ofdifferent products produced posesa challenge for the microbiologicalquality control as they are alternatelyproduced on the 8 production lines.Therefore, a reliable detectionsystem is indispensable for the dailyproduct release decisions for theproduction batches. AdelholzenerAlpenquellen permanently makesefforts to maintain the consistenthigh quality of the products byexpanding the quality control proces-ses in place. This is demonstrated,

for example, by the establishment of a new laboratory that meets the most modernrequirements and standards.

Routine microbiological analysis

The microbiological quality control processat Adelholzener Alpenquellen is characte-rised by a strict control of the finishedbeverages. Samples are taken, in definedintervals, from every production batch and then analysed. Accompanying this, a representative control process is carried outincluding the areas of raw materials, waterquality, beverage preparation and packagingdisinfection. The philosophy of the qualitycontrol process at Adelholzener Alpen-quellen includes, in addition to the analysisof finished products, established hygienemanagement processes and adaptedproduction management. Product safety and quality is assured due to the useof exact hygiene process cataloguesand product-alternation-matrices for allproduction lines, whereby every procedureand scenario is standardised (as far aspossible) and communicated within theplant. In this way, Adelholzener Alpenquellenachieves a product quality that goes farbeyond the mere analysis of the finishedproducts.

Microbiological quality control at Adel-holzener is split into two categories: the analysis of mineral water and theanalysis of sweetened drinks. Mineral wateris tested according to German regulations,in particular for the presence of E. coli andcoliform bacteria. Sweetened drinks aretested for the presence of beverage-spoilingmicroorganisms such as fermentativeyeasts, which represent the highest risk.

The analysis of E. coli and coliforms inmineral water takes up to 3 days (see Fig. 1).An important parameter for product releaseis the analysis of the lactose broth samples,which contributes significantly to the requirement of a 30 hour long quarantineperiod. The detection of fermentative yeaststakes between 3 days (for carbonateddrinks) and 5 days (for still drinks) (see Fig. 4), whereby certain quarantineperiods result.

VIT as an important part ofa reliable microbiological qualitycontrol process

In order to optimise the microbiologicalquality control process, the rapid detectionsystem VIT (vermicon identificationtechnology; vermicon AG, Munich) wasintegrated into the routine analysis for thedetection of E. coli and coliforms, as well as fermentative yeasts. The VIT method isbased on so-called gene probe technology.

20 · BBII 2/2006

A) Phase contrast of the sample B) Detection of living coliform bacteria incl. E.coli (red cells)

C) Detection of living E.coli cells(green cells)

Fig. 2: Analysis of the results of a VIT-E.coli/Coliforms test using a fluorescence microscope. Only some of the cells visible in the phase contrast are specifically identified as coliforms (including E.coli) – light up in red. Some of the coliforms are furtherspecifically identified as E.coli cells – light up in green.

A) Phase contrast of the sample B) Detection of living highlyfermentative spoilage yeasts (red cells)

C) Detection of living opportunisticspoilage yeasts (green cells)

Fig. 3: Analysis of the results of a VIT-Spoilage Yeasts test using a fluorescence microscope. Only some of the cells visible inthe phase contrast are specifically identified as highly fermentative spoilage yeasts – light up in red. Other cells are specificallyidentified as opportunistic spoilage yeasts – light up in green.

Gene probes marked with afluorescent dye penetrate the cellsin the sample and bind to specificsignatures. These signatures areonly present in living, active cells.Dead cells do not contain thesignatures. Therefore, the VITmethod reliably differentiatesbetween dead and viable microo-rganisms. The VIT test takes 3hours to be carried out, wherebythe hands-on time involved amounts to only a few minutes. The resultsare analysed using a fluorescencemicroscope. The target cells lightup in red or green whilst all othercells present in the sample do not light up (see Fig. 2 and 3). VITcan be used to analyse colonies or,for quicker results, the detection of microorganisms in a brothenrichment. Due to the simpleprocedure and the objectiveanalysis of the results (if cells lightup the result is positive; in not, the result is negative) VIT can beeasily integrated into the routineanalysis process.

E.coli/Coliforms:

The test VIT-E. coli/Coliforms is used to quickly and reliablyanalyse mineral water for the

presence of E. coli and coliformbacteria. If the lactose brothundergoes a suspicious colourchange after 24 hours of cultivation,5 bottles of the same filling timeare given a complete microbiologicalcheck. Should one or more samplesshow suspicious characteristics,the sample will be tested using VIT.Parallel to this, the results of theAPI test carried out on the originalsample are analysed. Only when a negative result is confirmed byboth API and VIT, the product isreleased. The fact that the resultsare available after only 24 hoursleads to a time-saving of 2 days in comparison with conventionalanalysis methods. All VIT analysesto date have shown 100 %compatibility with the results of the conventional method.

Spoilage Yeasts:

VIT-Spoilage Yeasts allows for thegeneral detection of living yeastsbut also the specific detection of both highly fermentative andopportunistic spoilage yeasts.Highly fermentative spoilageyeasts (genus Saccharomyces)are very likely to damage the taste of the product and in the

worst case may cause the productpackaging to burst due to the build up of CO2 gas. Opportunisticspoilage yeasts (several othergenera) can also damage products,whereby this heavily depends onthe individual characteristics of theproduct. Still drinks are generallymore at risk of product damage dueto the presence of opportunisticspoilage yeasts than carbonateddrinks.

The test for fermentative yeasts,VIT-Spoilage yeasts, is used toanalyse finished products as wellas samples out of the productionprocess. In the case of growth of a suspicious colonies, the VITtest delivers specific and reliableresults within 3 hours. The abilityto distinguish between highlyfermentative and opportunisticspoilage yeasts is particularlyadvantageous. In all of the trials carried out before the testwas implemented in the routineprocess, it was observed thatopportunistic spoilage yeasts didnot cause spoilage in carbonatedbeverages or beverages containingO2. Therefore, the products couldbe released. Thus, the reliabledifferentiation between the two

BBII 2/2006 · 21

Fig 4: Schematic representation of the analysis of spoilage yeasts in sweeteneddrinks as a part of the quality control process at Adelholzener Alpenquellen.

types of spoilage yeasts providedby the VIT method contributessignificantly to making decisionsregarding the release of products.

The practical relevance

A real example out of the routineanalysis shows how significant the differentiation between highlyfermentative and opportunisticspoilage yeasts can be: the basicmicroscopic analysis of a preparedstreak showed a strong similarityto Saccharomyces. The analysis of the colony using VIT-SpoilageYeasts revealed, within only a few hours, that it was not a highlyfermentative spoilage yeast of thegenus Saccharomyces, but ratheran opportunistic spoilage yeast.Observation of the yeast afterspiking beverage samples showedthat the yeast was not able to grow in the carbonated beverage.This confirmed that the yeast inquestion was definitely not a highlyfermentative spoilage yeast of thegenus Saccharomyces, exactly ashad already been revealed by theVIT test. Due to the unequivocaldifferentiation between highlyfermentative and opportunisticspoilage yeasts, the VIT testcloses a critical hole in the currentanalysis methods and increasesthe reliability of product releasedecisions.

Conclusion

Adelholzener Alpenquellen havethe clear aim, also with regards

to quality control, to further buildupon their leading position in thebeverage industry. The establish-ment of a new laboratory that meetsthe most modern requirements andstandards can be regarded as amilestone. The orientation towardsmodern analysis methods such as VIT also serves to achieve this aim.

Adelholzener Alpenquellen GmbHdecided to implement VIT as animportant part of the company’squality control process because of the faster analysis times, butalso because of the reliability of the results obtained using thedetection system. It was importantto have a detection system that provided timely results but could also provide reliableinformation about the actualbeverage-spoiling risk present.

It was also important that thedetection method could bestandardised and was notdependant on the expertise ofindividual employees. In addition,the method had to be able to beeasily integrated into the existingmicrobiological quality controlroutine. The VIT method fulfilledthese requirements and wastherefore, after an internalvalidation phase, implemented as a part of the routine analysis. In practice it has been shown that the VIT method contributessignificantly to the quality controlprocess due to the reliable andunequivocal results that it delivers.

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Rapid detection systems for microorganisms

Drinks


Drinks

VIT-Alicyclobacillus Art. No. 01210013

VIT-Alicyclobacillus detects all bacteria of the genus Alicyclobacillus and all bacteria of the

species Alicyclobacillus acidoterrestris at the same time in one test. Only living bacteria are

detected. Even the spores of these bacteria can be detected in all stages of the production

process for fruit juices and fruit juice concentrates. The test is carried out after a short pre-

cultivation of the sample. The hands-on time required per test is only a few minutes and the

results are available within 3 hours. The results are analysed using a fluorescence

microscope. A time saving of up to ten days is realised in comparison to conventional

methods.

VIT-Spoilage Yeasts Art. No. 01210016

VIT-Spoilage Yeasts reliably detects all living fermentative yeasts in still and carbonated

drinks. Additionally highly fermentative spoilage yeasts can be separately identified from

opportunistic spoilage yeasts. Highly fermentative spoilage yeasts quickly lead to a

deterioration of the product. The possibility of deterioration due to opportunistic spoilage

yeasts depends on the characteristics of the product. The test is carried out after a short pre-

cultivation of the sample in a suitable medium. Suspicious colonies can also be tested. The

hands-on time required per test is only a few minutes and the results are available within 3

hours. The results are analysed using a fluorescence microscope. A time saving of up to four

days is realised in comparison to conventional methods.

VIT-Lactic Acid Bacteria Art. No. 01210018

VIT-Lactic Acid Bacteria detects viable lactic acid bacteria belonging to Lactobacillus,

Pediococcus and Leuconostoc in beverage samples. The test is carried out after a short pre-

cultivation of the sample or grown colonies can be analysed. The hands-on time required per

test is only a few minutes and the results are available within 3 hours. The results are

analysed using a fluorescence microscope. A time saving of up to 2 days is realised in

comparison to conventional methods.


Drinks

VIT-Bier plus L. brevis Art. No. 01210006

VIT-Bier plus L. brevis detects all beer-spoiling lactic acid bacteria and the most frequently

occurring beer pest Lactobacillus brevis at the same time in one test. Only living bacteria are

detected. The test is carried out after a short pre-cultivation of the sample. The hands-on

time required per test is only a few minutes and the results are available within 3 hours. The

results are analysed using a fluorescence microscope. A time saving of approximately four to

five days is realised in comparison to conventional methods. The test can be carried out in all

steps of the brewing process – filtered area, unfiltered area, yeast store.

VIT-Bier Megasphaera/Pectinatus Art. No. 01210007

VIT-Bier Megasphaera/Pectinatus detects all bacteria belonging to the genus Pectinatus and

all bacteria belonging to the species Megasphaera cerevisiae at the same time in one test.

Only living bacteria are detected. The test is carried out after a short pre-cultivation of the

sample. The hands-on time required per test is only a few minutes and the results are

available within 3 hours. The results are analysed using a fluorescence microscope. A time

saving of approximately five to eight days is realised in comparison to conventional methods.

VIT-E. coli/Coliforms Art. No. 01210008

VIT-E. coli/Coliforms detects all coliforms and all E. coli strains at the same time in one test.

Even lactose- and indol-negative E. coli strains are detected. Only living bacteria are

detected. The test is carried out after a short pre-cultivation of the sample. The hands-on

time required per test is only a few minutes and the results are available within 3 hours. The

results are analysed using a fluorescence microscope. A time saving of approximately one to

two days is realised in comparison to conventional methods.

safety for beverage production -...

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