Dairy Products Microbiology TestingProven solutions and services for every step of your microbial testing workflow.

The life science business of Merck operates as MilliporeSigma in the U.S. and Canada.

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We provide products and services to support every step of your dairy product microbio-logy testing process. Our microbial testing products and kits comply with international standards; our portfolio includes dehydrated and granulated culture media, ready to-use agar plates, liquid media, as well as molecular (PCR) or immunological solutions for the specific detection of pathogens such as Listeria, Salmonella, Campylobacter or E. coli or the detection of quality indicator organisms. We also have a comprehensive range of services and high-quality environmental monitoring solutions to help keep your facilities safe for convenient checking of surfaces, equipment, and air quality for contaminants – for rapid, reliable results and international standard compliance.

Our expertise in the food industry As a worldwide leading provider of life science solutions, we address the needs of customers in industries where consumer safety is a major concern. Our experts are dedicated to ensuring reliable safety testing results.

Bringing wholesome and safe products to market fast We provide dependable, connected and rapid solutions specifically tailored to the needs of the food and beverage industry.

What makes us different? We offer a complete portfolio with proven solutions & technologies that are compliant to international standards throughout the entire safety testing workflow.

We are setting standards in the food industry

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Dairy Safety & Testing Solutions

SigmaAldrich.com/Food-Microbiology

The Importance of Dairy Testing

Throughout your dairy production process, specific tests should be carried out to ensure the quality and safety of your final dairy products from milk, yogurt, cheese, ice cream and dried milk to infant formula.

Milk and dairy products provide a favorable environment for a large range of bacteria, yeasts and molds; it is therefore important to monitor these organisms to help prevent spoilage or potential pathogenic contamination.

Our range of solutions cover the entire production process through to the QA/QC analysis of your final products while ensuring compliance with regulatory standards.

How We Test

Dairy processing, manufacturing and quality control requires testing of specific microorganisms. Common pathogenic contaminants in dairy include Salmonella, Campylobacter, Listeria, E. coli, S. aureus, and Cronobacter amongst other spoilage organisms.

Your microbiological testing program needs to take a number of points into consideration, such as the frequency of testing, which stages in the production process to carry out testing, the matrix or product composition, which target organisms to test for, and which test methods to use.

How We Can Support Your Testing Program

Our comprehensive range of QA/QC microbiological solutions:

• Environmental monitoring for microbiological analysis of air, surfaces and personnel

• Bioburden testing

• Pathogen testing

• Indicator organism testing

• Spoilage organism testing

• Allergen testing

• HACCP management and ATP testing

• Services - from method development to maintenance

Tests range from quantitative and qualitative depending on your needs. If you are following specific regulatory methods, we can provide the right solutions for your protocol, along with a selection of rapid methods.

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Focus: Infant FormulaInfant formula and their ingredients are foods that have specific testing needs. It is critical to verify the absence of Cronobacter spp. and Salmonella in powdered infant formula (PIF). While Salmonella has the capability to infect all humans, Cronobacter spp., formerly Enterobacter sakazakii, is a pathogenic bacterium that can cause severe disease, primarily in infants under 6 months old and immunocompromised adults. This organism can survive in low moisture foods, such as PIF, for up to two years.

In the EU, Commission Regulation (EC) No. 2073/2005 requires the absence of Cronobacter, Enterobacteriaceae, and Salmonella in dried infant formula and dried dietary foods for special medical purposes intended for infants below six months of age. This food must also be tested for presumptive presence of Bacillus cereus. In the US, all finished dried product infant formula samples (domestic or import) designated for microbiological analysis, need to be tested for Salmonella and Cronobacter. For Salmonella analysis, each sample should consist of 25 grams or larger, for Cronobacter analysis it is 10 grams or larger.

Most countries have standards for quality control (QC) of PIFs and their ingredients. QC labs of food companies exporting PIF need to be aware of the food testing regulations in the countries of destination. Culture media-based methods are described in all countries, but a careful check of the compliance of culture media products used in quality control is important. The culture media should be of high quality and should be accompanied by clear documentation about the compliance to the given standards. This compliance information should be visible in both the product and batch documentation provided by the media supplier. Such documentation also supports QC managers in audit situations and lab inspections.

As described above, a rapid method can only be applied if permitted in the country of destination. These methods are only acceptable if validated according to ISO 16140, AOAC International standards or a specific local standard if needed. Molecular methods for detection of Cronobacter and Salmonella are preferred as they can detect these organisms in low numbers. A molecular system needs to provide excellent performance data of the individual system with PIF as food matrix. A sample preparation procedure that efficiently extracts the target nucleic acid is important, e.g. by magnetic separation which leads to higher test reliability.

Read page 18 for more information.

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ProductRaw milk & derivatives

Pasteurized milk

Dairy products

Powder milk

Infant formula

Total count (aerobic mesophilic bacteria)

Somatic cells

E. coli

Coliforms

Clostridium perfringens

Enterobacteriaceae

Cronobacter

Salmonella

Listeria monocytogenes

Thermotolerant Campylobacter spp.

Bacillus cereus

Coagulase positive Staphylococci

Staphylococcal enterotoxin

Molds

Probiotics

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Which are the most important microorganisms in dairy products?Milk and dairy products need to be examined for diverse set of organisms depending on the product type and the country. The typical testing specifications per product type are listed below.

Total aerobic, mesophilic and somatic cell counts give information about the quality and freshness of milk along with hygiene and animal health. E. coli, coliforms, C. perfringens and Enterobacteriaceae are important indicators for fecal contamination. Cronobacter, Salmonella, Listeria monocytogenes, thermotolerant Campylobacter, Bacillus cereus, and coagulase-positive staphylococci are possible pathogens in milk and

dairy products. Most of them can produce toxins like staphylococci and Bacillus cereus. Molds can also produce dangerous toxins, but the regulations often only state that there should be no molds visible by eye. Probiotics are beneficial organisms for the human body and are commonly used in dairy products and infant formula.

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Enterobacteriaceae

Enterobacteriaceae are intestinal bacteria which can be found in dairy products from contamination of food, insufficient pasteurization and sterilization or recontamination due to inadequate hygiene standards. We offer diverse media and tests, like oxidase tests, for the detection and enumeration of Enterobacteriaceae.

Cronobacter (Enterobacter sakazakii)

Cronobacter is a Gram-negative, facultative anaerobic rod-shaped and motile bacterium that belongs to the Enterobacteriaceae family. The microorganism can cause severe neonatal infections: necrotizing enterocolitis, septicaemia and meningitis. The fatality rate following meningitis and other infections is 50%, with the survivors being neurologically damaged for life. Fortunately, infections are rare in infants, but they can occur in all age groups, admittedly with less severe clinical outcomes. Cronobacter spp. can grow over a wide temperature range, between 5.5 °C and 8 °C the multiplication begins. Average generation times of Cronobacter is about five hours at 10 °C and only 40 minutes at 23 °C. The organism is very tolerant of drying steps and can survive for two years desiccated in infant formula and then rapidly grow on reconstitution.

Escherichia coli & Coliforms

Coliforms are intestinal bacteria that are often present as a result of contamination by feces. They are defined as rod-shaped Gram-negative facultative anaerobic non-spore forming organisms that ferment lactose with the production of acid and gas when incubated at 35–37 °C. In most cases coliforms do not cause illness, but they are used as indicators for other pathogenic organisms of fecal origin that may be present in foods and water. Escherichia coli (E. coli): Is the most famous member of the coliform group bacteria and is an important indicator for fecal contamination, as it is almost exclusively of fecal origin.

We offer a broad range of tests and culture media, also chromogenic and fluorogenic media, for detection of E. coli and coliforms.

SigmaAldrich.com/Ecoli

Clostridium perfringens

C. perfringens is also a fecal indicator organism but is much more resistant than E. coli and, due to the ability

to produce spores, can be found even after treatments like pasteurization. We offer our unique chromogenic CP ChromoSelect agar but also diverse other media and tests to detect this organism.

SigmaAldrich.com/Clostridium-Article

Salmonella

Salmonella contamination is the second leading cause of foodborne illness worldwide. Controlling outbreaks of Salmonella is an important task for the food industry. The Salmonella family includes over 2,300 serotypes of bacteria, but two types, Salmonella Enteritidis and Salmonella Typhimurium, are responsible for about half of all human infections. Most outbreaks of Salmonella are traced back to dairy, poultry and meat products. Salmonella have the necessary tools and mechanisms to spread in the body.

Microbiological control in the food industry plays a critical role in preventing Salmonella outbreaks and is probably an important reason for decreasing incidences in recent years. Tests and media used for identification of Salmonella take advantage of unique aspects of Salmonella physiology or biochemistry relative to other genera within the Enterobacteriaceae family. For example, bacteria from the genus Salmonella are mostly facultative anaerobes, oxidase-negative, catalase-positive and Gram-negative rods. Most strains are motile and ferment glucose with production of both acid and gas.

Learn more about Salmonella contamination: SigmaAldrich.com/Salmonella-Article

Listeria monocytogenes

In recent years, it has been recognized that Listeria monocytogenes is an important public health problem. Listeriosis primarily affects people of advanced age, pregnant women, newborns, and adults with weakened immune systems. The infection increased over the last few years, most likely because of an increasing variety of food products, the trend for “ready-to-eat” and “ready-to-cook” products plus the demand for longer shelf life. Therefore, it is even more important that the risk foods, like dairy products, are thoroughly checked for contamination.

Read our article about Listeria monocytogenes: SigmaAldrich.com/Listeria-survivor

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Yoghurt sample cultured on BSM Agar. Bifidobacteria appears as purple-brown colonies.

Thermotolerant Campylobacter

Campylobacteriosis is currently one of the most common bacterial infections in humans and is often a foodborne illness. The two species, Campylobacter jejuni and C. coli, are responsible for the majority of human foodborne campylobacteriosis. There are other pathogenic species—C. lari, C. fetus (infect reproductive tract), C. upsaliensis and very seldom C. helveticus—which can also cause diarrheal illness. Campylobacter can occasionally be found in raw milk. They are microaerophilic organisms that require oxygen levels between 2 and 10% for growth. These two species are classed as thermophilic campylobacters as they have an optimum temperature for growth of 42 °C. The current method used for detection of campylobacters from foodstuffs is the EN/ISO 10272-1:2017 method.

SigmaAldrich.com/Campylobacter-Testing

Bacillus cereus

Bacillus cereus is a typically motile soil bacterium which is able to build resistant centrally located endospores. The colonies that grow on most media types are dull and frosted. It is a Gram-positive and a facultative anaerobic organism, but older cells may become Gram-negative as they age. The Bacillus cereus group comprises Bacillus anthracis, Bacillus thuringiensis, Bacillus cytotoxicus and some other species, which share about 3100 genes. Some strains can cause foodborne illness similar to those caused by Clostridium perfringens or Staphylococcus aureus. The minimum infective dose is about 106-107 bacteria/g, at that range of cell concentration they start to build different toxins which cause both diarrheal and emetic human gastrointestinal syndromes. One of the infection sources for the diarrhea type of B. cereus infection are UHT milk products. Contamination of raw milk occurs by simple transfer during milking when hygiene conditions are not fully observed. If present in raw milk, spores can survive the pasteurization process and contaminate the finished milk product.

In milk products, B. cereus and other bacilli can degrade the casein to peptides which may be further degraded to amino acids which are responsible for alkaline reaction and a bitter taste in milk. The ISO recommends a colony count technique ISO 7932:2004 and an MPN method ISO 21871:2006 to detect and enumerate B. cereus. Coagulase-positive Staphylococci

Coagulase-positive staphylococci comprise Staphylococcus aureus, Staphylococcus hyicus and Staphylococcus intermedius. S. aureus is reported to be the most pathogenic species and is responsible for food poisoning due to the production of several relatively heat stable exotoxins. Enterotoxins are produced if the colony forming units per g sample exceeds 105 cfu/g. While in most cases for dairy products for S. aureus cells, a concentration of up to 100 cfu/g is acceptable, the toxin should not be detectable.

Read more: SigmaAldrich.com/Staph-Aureus

Probiotics

Probiotics are defined as live bacteria with beneficial effects to the health of the host organism. Probiotics take on a key role in the functional food industry. Today, lactic acid bacteria (LAB) and Bifidobacteria are primarily used as probiotics, however certain yeasts and bacilli are also known to have positive effects. In most cases, probiotics are produced directly by fermentation in foods such as yogurt or are supplied through dietary supplements.

Bifidobacterium is one of the most important probiotic bacteria used in the dairy industry. Bifidobacteria have a positive effect on the immune system and control intestinal pH. Additionally, they produce bacteriocins and bacteriocin-like inhibitory compounds which inhibit the growth of other bacteria. ISO 22981:2010 describes the enumeration of presumptive Bifidobacteria by the TOS-MUP agar. Our 100043 TOS-MUP agar is in accordance with this ISO standard and is highly selective for Bifidobacteria as it suppresses the growth of Lactobacilli. Our chromogenic Bifidobacteria Selective Media allows for easy and fast quality control of yogurt: SigmaAldrich.com/Bifidobacteria-Media

Lactobacilli belong to the lactic acid bacteria and comprise the major part of this group. As their name implies, they produce lactic acid. The acid-producing mechanism inhibits growth of other organisms and favors the growth of Lactobacilli that thrive in low pH environments. Some strains of Lactobacilli were shown to produce, like Bifidobacteria, a bacteriocin-like substance and can inhibit a broad range of pathogens.

Probiotic Species Product Name Company

Bifidobacterium breve Yakult Bifiene Yakult

Lactobacillus casei DN114-001 Actimel/DanActive Danone

Lactobacillus casei Shirota Yakult Yakult

Lactobacillus johnsonii La1 LC1 Nestlé

Table 1: Some famous probiotic products

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SigmaAldrich.com/Microbial-SamplePrep

Food Sample PreparationAccurate microbiology results are highly reliant upon the quality of your sample preparation.

Sample DilutionOur DiluCult™ automated gravimetric dilutor instruments with two integrated pumps are designed to facilitate work in a microbiological safety cabinet for samples of up to 5 kg. They are low in height and have a removable drip tray. Three modes of dispensing are available: a fast flow with minimum accuracy, a standard flow with standard accuracy, and a slow flow with optimal accuracy. Our DiluCult™ and DiluCult™ 2 instruments can be easily calibrated with a certified weight.

Food Sample Homogenizer (Blender)The Enrichment Sample Homogenizer (ESH) is used for the automated homogenization of food samples with enrichment media, the ESH has two removable and autoclavable massaging paddles for straightforward cleaning.

• Multi-function digital display & control panel• Variable speed and time• Adjustable blending power• Side-by-side paddle stop• Silent, brushless motor • Security drip tray• Removable autoclavable paddles

SigmaAldrich.com/DCM

Watch our video

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Enrichment and Cultivation

Dehydrated Culture MediaThe superior granulation technology of our traditional dehydrated culture media meets the highest industry performance standards, while guaranteeing that our products provide maximum convenience and safety.

GranuCult® ISO 11133: 2014 Compliant Granulated Culture Media

Our GranuCult® low-dust dehydrated culture media granules are compliant with EN ISO 11133:2014 standards, as well as other ISO standards and FDA-BAM and USDA-FSIS methods for food and water sample testing. Our GranuCult® media are released through an ISO 17025 QC lab. Reagulytory compliance is clearly visible on the product label and all test strains are listed in the certificate of analysis.

Ready-to-Use Culture MediaOur prepared and ready-to-use culture media products can save you time and resources, while ensuring that your media is fully compliant with the regulations relevant to the dairy industry. Whether you need carry out pathogen testing or indicator organism testing in food or beverage samples, we have a full range of liquid media and pre-filled agar plates suitable for your needs.

ReadyTube® Liquid Media

Compliant with EN ISO 11133:2014, our ready-to-use media in tubes and bottles for isolation, enumeration or enrichment of microorganisms. The clear naming convention where the number indicates the volume in mL of media contained in each bottle or tube makes it simple (e.g. ReadyTube® 200—each bottle in the pack contains 200 mL of media).

Certified Reference Microorganisms

Vitroids™ and LENTICULE® Discs Take the hassle out of your culture media performance testing and quality control. Our range of Vitroids™ and LENTICULE® Discs are certified reference materials that can help make your quality control tests quick, convenient and consistent. Provided as ready-to-use discs in convenient CFU levels, there is virtually no prep needed: simply place the disc onto or into the media of your choice, rehydrate for 10 minutes, and process as normal. It’s as easy as that.

Streamline your media performance testing and daily quality control:

• Ready-to-use at convenient CFU ranges• No dilution required • Quick rehydration in 10 minutes• 1+ year shelf life in standard -20 °C freezer• Complete with comprehensive certificate of analysis • Double accreditation to ISO 17034 and 17025

Use for media performance testing, method development and validation, process quality control and for employee training and proficiency testing.

Cultivation of Anaerobic BacteriaMany microorganisms involved in food and beverage contamination are anaerobes, microaerophiles or capnophiles such as Clostridium, Campylobacter or Lactobacillus. Our Anaerocult® systems produce the anaerobic, microaerobic and capneic growth conditions that are required for cultivation of these microorganisms.

Sample AnalysisPathogen detection

SigmaAldrich.com/GDS

SigmaAldrich.com/DCM

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Traditional MethodsWe provide a broad selection of high-quality culture media to detect pathogenic bacteria in your dairy products. From dehydrated culture media to ready-to-use media, we have a solution to match your needs.

SigmaAldrich.com/Transia

ReadyBag® Ready-to-Use Media

Our pre-weighed and gamma-irradiated Readybag® granulated media pouches speed up and simplify your food pathogen testing routines. With our single-use Readybag® Buffered Peptone Water and Half FRASER broth pouches, there is no weighing and no autoclaving. Readybag® Half FRASER Broth even eliminates the need to prepare and sterile filter the supplements because they are already incorporated into the medium, saving you even more time and expense.

Rapid MethodsRapid methods, if not described in specific USDA, FDA, ISO, or any other standard are considered as alternative methods that are preferred if short time to result is needed to release food samples for human consumption. Alternative methods are applicable if validated according ISO 16140 and/or AOAC International guidelines. The acceptance of which can be dependent on your geographical region.

PCR Detection

Assurance® GDS combines the latest advancements in molecular detection technology and food microbiology to provide faster results with the increased accuracy required to meet today’s food and environmental testing challenges.

Our newest assay, Assurance® GDS Cronobacter Tq II has been developed to meet the needs of laboratories testing for Cronobacter spp. in infant formula, infant cereals and environmental samples. The system uses the targeted isolation capabilities of PickPen™ Immunomagnetic Separation (IMS) together with the specificity of DNA-based detection, to provide reliable results in as little as 22 hours.

In addition, the Assurance® GDS pathogen detection system includes validated assays (AOAC / MicroVal / Health Canada / AFNOR) for Salmonella, E. coli O157:H7, Non-O157 Top STEC, Shiga Toxin Genes, Listeria spp., and Listeria monocytogenes.

Enzyme-linked Immunosorbent Assay (ELISA) Method

TRANSIA® AG and TRANSIA® PLATE incorporate proven antibody-antigen “sandwich” assay technology for the detection of food borne pathogens in both product and environmental samples. The TRANSIA® Salmonella Gold and TRANSIA® PLATE Listeria are fully ISO 16140-2 validated.

How it works: microtiter plate wells coated with highly specific antibodies capture and bind the target antigen if present. A detection antibody linked to a conjugate enzyme is then introduced to form an antibody/antigen/antibody sandwich and finally, a sub-strate is added and converted by the conjugate enzyme to produce a color change indicating the presence of the target pathogen. The Gemini instrument offers the option of a complete automation, from pipetting, washing and incubating to reading and interpretation.

SigmaAldrich.com/Lateral-Flow

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SigmaAldrich.com/IDMembranes

Lateral Flow Tests

Lateral Flow Tests are immunoassays for detecting food pathogens with the ultimate convenience.

Singlepath® & Duopath® Lateral Flow Tests Covering the major pathogens, these tests act as mini-laboratories and include a built-in control reaction for optimized pathogen testing. Definitive results are delivered in as little as 20 minutes. Use in combination with our GranuCult® dehydrated media to ensure optimal test performance.

Benefits:

• Reliable: same accuracy standards as classical detection methods

• Comprehensive: cover the most relevant pathogens in food – Campylobacter, E. coli 0157, STEC, Salmonella (all AOAC-RI approved), Listeria monocytogenes and Bacillus cereus Enterotoxins.

• Fast: Definitive results within 20-30 minutes

• Easy-to-use: Clear yes/no results after simple sample application

• Safe: Additional positive control and specially adapted enrichment media for reliable results

VIP® Gold Lateral Flow Tests VIP® Gold (Visual Immuno Precipitate) is a lateral flow immunoassay for the detection of pathogens in food and environmental surfaces. Each VIP® Gold device is a self-contained test that requires only the addition of sample, making it an extremely simple and user-friendly method ideal for low to medium volume labs. VIP® Gold tests are AOAC and Health Canada approved and available for testing:

• Salmonella • E. coli O157:H7• Listeria spp.

ID Membranes for Rapid Identification or Confirmation

Agar plates are often used for the detection or enumeration of microorganisms. Our smart, inexpensive and simple ID membranes can help to identify or confirm organisms within 1 to 4 hours, directly from your plates. After routine inoculation and isolation, these membranes enable the direct identification based on chromogenic and fluorogenic substrates. Simply The place the membrane on the colonies grown on the plate, then after a short incubation time, the colors or fluorescence develop in the presence of specific microorganisms.

We offer a broad range of different ID membranes to identify diverse organisms from water, food, environmental and clinical samples.

Microbiological Identification and Confirmation

Indicator organism testing

Contamination prevention is the key to keeping our food supply safe. It is therefore necessary to continuously monitor food production facilities and final products for indicator organisms. These organisms indicate potential hazards in the finished products, which can lead to costly product recall.

Traditional MethodsThere are different ways to test for quality indicators and / or index organisms, including traditional methods using our gold standard GranuCult® dehydrated media, or our ready-to-use solutions, for example ReadyPlate™ agar plates or ReadyTube® liquid media.

ReadyPlate™ Agar Plates

For isolation and enumeration of indicator organisms, our ReadyPlate™ prepared agar plates are certified to be fully compliant with EN ISO 11133:2014, as well as with individual standards, and are quality controlled by ISO/IEC 17025:2005 accredited laboratories. Each plate is provided with a label including a data matrix code for paperless plate identification.

Our range includes:

• ReadyPlate™ (90 mm plates) e.g. XLD agar, MYP, BPA, PEMBA etc.• ReadyPlate™ CHROM (90 mm plates) Chromogenic media e.g. CCA, TBX and Listeria Agar

Ready-to-use Liquid Media: ReadyTube® Media

Discover ready-to-use media in tubes and bottles for isolation, enumeration or enrichment of microorganisms in food. Our ReadyTube® bottles and tubes are fully compliant with EN ISO 11133:2014 and as well as with individual standards, and are quality controlled by ISO/IEC 17025:2005 accredited laboratories to ensure compliance.

Our clear naming convention where the number indicates the volume in mL of media contained in each bottle or tube (e.g. ReadyTube® 200 media where each bottle in the pack contains 200 mL of media) makes it simple.

• Easy visibility of compliance on product label and certificate of analysis• Safe testing with maximum convenience• No time-consuming preparation• High reproducibility with a low error rate• Long shelf life of 6-12 months• Released through ISO 17025 accredited QC lab for reduced incoming quality control tests

SigmaAldrich.com/RTU-Media

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SimPlate® Tests

When working with complex matrices, our SimPlate® device can help make testing simple. This ISO and AOAC validated assays provides rapid, easy to interpret results even with food matrices that are challenging for other rapid methods. The SimPlate® system with Binary Detection Technology™ represents the latest technological advancement in counting techniques for the food industry. Results are presented as a clear visual positive or negative readout without the need for subjective interpretation, which provides consistency across users, generating more reproducible results.

Accurate results in four steps

Alternative methodsOur range of rapid culture media plates are an efficient method for the detection and enumeration of microorganisms including quality indicators, pathogens and spoilage organisms. They are also suitable for testing of difficult matrices.

MC-Media Pad®

Use fewer handling steps for your indicator organism testing, with our MC-Media Pad® ready to use method. This convenient method complies with international food and beverage standards (AOAC-PTM & MicroVal, ISO 16140), and it can be used for convenient microbiological testing of coliforms and E. coli, yeast and mold, or aerobic microbial contamination. The clear color coding means that you will always pick the right one at a glance. The MC-Media Pad® improves your workflow and reduces required storage, incubation and waste capacity. Simply inoculate your sample, incubate, and count your results!

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Open the cover film diagonally

2. Inoculate your

sample, it will diffuse automatically

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Close the cover film and incubate

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Count your results

SigmaAldrich.com/Alternative-Media

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SigmaAldrich.com/Microbial-Air-Monitoring

SigmaAldrich.com/Surface-Monitoring

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Hygiene Monitoring

Effective cleaning and disinfection, the foundations for good hygiene, have been a product focus and passion of ours for many years. Whether it is the environment, the equipment, the surfaces, air, water or wastewater, we have the solutions.

Our hygiene monitoring range has been developed and optimized in close cooperation with industrial users and operators, as well as with opinion leaders of supervising authorities.

Active and Passive Air MonitoringHygiene standards in a production environment are directly linked to the microbiological safety of the finished products. We therefore make sure that our active microbial air sampling solutions generate highly precise, reliable and reproducible results. We offer a comprehensive range of active and passive microbial air monitoring solutions for effective use in the food and beverage industry where regulations are on the rise, and standards become increasingly stringent.

Our air samplers are based on different principles, enabling selection of the technology to suit your needs. Our range includes MAS-100® air samplers which are based on the Anderson impaction principle that use settle plates or contact plates, as well as RCS® (Reuter Centrifugal Sampler) that use dedicated agar strips. There are product variants specifically designed for economical use in the food and beverage industries, for explosion hazard areas and for monitoring of compressed gases.

MAS-100® ECO Air Sampler

The MAS-100® ECO microbial air sampling system is designed for reliable and easy microbial monitoring of ambient air in food and beverage manufacturing facilities.

• Accurate, reproducible measurements and outstanding user flexibility• Easy yes /no button operation• Built-in airflow compensation and fully automatic calibration• Robust and designed to operate in food manufacturing environments

Passive Air Monitoring: LI (Long Incubation) Settle Plates

LI Settle Plates are single-bagged prepared media ideal for use as settling plates. These plates contain either Tryptic Soy agar (TSA) general purpose media, or Sabouraud Dextrose agar (SDA), which is ideally suited for the growth of yeast and molds.

• Higher than standard filling volume of 30 mL to compensate for water loss• Barcoded to enable full traceability

Traditional Surface and Liquid MonitoringWe provide a wide range of solutions for surface, personnel and liquid testing, which are critical to ensure that your production plants are clean, and the contamination risk of products and processes is minimized. For monitoring the efficiency of disinfection routines, proven methods and easy-to-use testing solutions are available for standard flat and curved surfaces as well as difficult to access areas. We offer a wide range of traditional agar media solutions such as ready-to-use contact plates, contact slides as well as swabs, each being suited to different surface types.

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Contact Slides, Dip & Swabs

Our Dip and Swab testers are designed for convenient microbial counting in a broad range of applications. Using the swab, you can reach even difficult to access areas. These ready-to-use dehydrated dip testers are available for heterotrophic plate count (HPC), yeast & mold, and coliforms. They are convenient and easy to use, suitable for liquid and swab samples.

Our Envirocheck® Dip Slides for semi-quantitative detection of microorganisms on surfaces and in liquids are flexible paddles that are coated with different nutrient media. After sampling and incubation, two parameters can be determined in one step. Envirocheck® Dip Slides are available for the following parameters: Disinfection control, Total Colony Count, Detection of yeasts and molds, Detection of Enterobacteriaceae and Detection of total coliforms / E. coli.

Rapid Surface MonitoringWe offer a range of solutions for rapid environmental monitoring disinfection control and HACCP management including convenient media, ATP testing and allergen and protein detection.

MVP ICON® Hygiene Monitoring and HACCP Management System

The MVP ICON® instrument and software platform combines HACCP and hygiene monitoring with powerful program management capabilities. The MVP ICON® system allows you to monitor key HACCP parameters, reducing the need for multiple instruments. Available measurements include:

• ATP (Adenosine Triphosphate)• Chemical Concentration (ppm)• Conductivity (µS)• pH• Temperature

Rapid results allow for real-time decision making and corrective action. The patented design of the sampling device ensures a high degree of accuracy—even in the presence of sanitizers. The MVP ICON® instrument provides the most complete HACCP and hygiene monitoring system with dashboard software that displays key performance metrics, so you can effectively manage your quality assurance program.

FLASH® Rapid Allergen Indicator Protein Detection Test

Protein swab tests are used to help quickly verify cleaning effectiveness by detecting protein residues, including allergens left on food contact surfaces after cleaning. The FLASH® total protein test gives a visual readout with results within 10 minutes, allowing corrective action to be taken immediately. No instrumentation is required, and the test can be used by all personnel without the need for extensive training.

• Single swab providing flexible test method based on desired level of sensitivity: – Room temperature readings detect down to 20 μg – Achieve higher sensitivity detection down to 3 μg when incubated at 70 ⁰C

• Detects total protein, including samples comprising the “Big 8” food allergens: gluten flour, soy flour, egg powder, milk powder, roasted almonds, peanut butter, raw shrimp, and raw fish (cod)

SigmaAldrich.com/Rapid-Surface-Monitoring

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Microbiology Services

Are you ready to start testing? Our Services support our customers in the food industry throughout the entire workflow of food safety testing in order to ensure optimum performance and regulatory compliance. Regardless of product, we offer a variety of services* including:

• Application services: feasibility studies using your specific matrices, method development and method verification

• Validation services: instrument qualification, PQ support, regulatory consultancy

• Maintenance services: calibration, preventive maintenance, repair, extended warranty on-site or in our local repair center

• Instrument service and preventative maintenance plans (Total Coverage, Total Service, Verification Service)

* Individual service offerings may vary per product and geography, ask your local representative for details

SigmaAldrich.com/Food-Services

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Let’s create regulatory compliance. Together.In the food industry, where consumer safety is a major concern, stringent standards regulate the production environments. The quality control of food products and their ingredients is regulated by international standards. Countries that don’t follow ISO, FDA, or USDA methods may have developed their own methods that are different, making it challenging to stay updated.

Visit our website to find regulatory resources that help you to stay compliant. We offer articles, whitepapers and webinars about international regulatory standards for free.

To find out more, visit SigmaAldrich.com/Food-Methods

What if safety were an open book?

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In ISO following countries, QC labs can choose to follow either culture media-based ISO standards or alternatively ISO 16140 validated rapid methods. ISO, the International Organization for Standardization, published revised culture media-based standards in 2017 for Salmonella (ISO 6579-1: 2017), Cronobacter spp. (ISO 22964:2017), and for Enterobacteriaceae (ISO 21528:2017), which are all relevant for testing of powdered infant formula and its ingredients.

The revised EN ISO 22964:2017 standard specifies a horizontal method for the detection of Cronobacter spp. in food, animal feed and environmental samples. It is applicable to food products and ingredients intended for human consumption and the feeding of animals, and environmental samples in the area of food production and food handling. In the new EN ISO 22964:2017, the scope has been extended to detect Cronobacter spp. according to new taxonomy from 2014. Cronobacter spp. are those microorganisms which form typical colonies on chromogenic Cronobacter isolation (CCI) agar and which display the described biochemical characteristics when tests are carried out in accordance with this new standard. To enable customers to follow EN ISO 22964:2017, we have released the newly described Cronobacter Selective Broth (CSB) and CCI agar products.

The revised EN ISO 6579-1 standard, which specifies a horizontal method for the detection of Salmonella spp. in the food production chain. Like the preceding ISO 6579:2002+Amd 1:2007, the new EN ISO 6579-1 covers the testing of products intended for human consumption, animal feeding and environmental samples in food production and handling. In addition, EN ISO 6579-1 now includes testing of milk and milk products as well as samples from the primary production stage, such as animal feces, dust and boot socks.

The revised EN ISO 21528 consists of 2 parts. In Part 1, a horizontal method for the detection of Enterobacteriaceae is specified, and in Part 2, the enumeration of these organisms, both in food and animal feed samples is stated. The revised standards are also applicable to environmental samples in the area of primary production, food production and food handling. Part 1 is applicable when the microorganisms sought are expected to need resuscitation by enrichment, and when the number sought is expected to be below 100 colony forming units (cfu) /mL or gram of test sample. Part 2 is applicable when the number of colonies sought is expected to be more than 100 cfu / mL or gram of the test sample. The new Part 1 edition cancels and replaces the first edition EN ISO 21528-1:2004. Part 2 cancels and replaces the first edition of EN ISO 21528-2:2004.

Food categoryMicro-

organisms

Sampling plan (20) Limits (21)

Analytical reference method (22)

Stage where the criterion

applies

Action in case of unsatisfactory

resultsm Mn c

2.2.1. Pasteurized milk and other pasteurized liquid dairy products (32)

Entero- bacteriaceae 5 2 <1 cfu/mL 5 cfu/mL ISO 21528-1

End of the manufacturing

process

Check on the efficiency of heat-treatment and prevention of reconta- mination as well as the quality of raw materials

2.2.2. Cheeses made from milk or whey that has undergone heat treatment

E. coli (33) 5 2 100 cfu/g 1 000 cfu/g ISO 16649-1 or 2

At the time during the manufacturing process when the

E. coli count is expected to be highest (34)

Improvements in slaughter hygiene and

review of process controls

2.2.3. Cheeses made from raw milk

Coagulase-positive

staphylococci5 2 104 cfu/g 105 cfu/g EN/ISO 6888-2

At the time during the manufacturing

process when the number of staphylococci is expected to be

highest

Improvements in production hygiene and selection of raw

materials. If values >105 cfu/g are detected, the cheese batch has to be

tested for staphylococcal enterotoxins

2.2.4. Cheeses made from milk that has undergone a lower heat treat- ment than pasteu-rization (35) and ripened cheeses made from milk or whey that has undergone pasteu- rization or a stronger heat treatment (35)

Coagulase-positive

staphylococci5 2 100 cfu/g 1 000 cfu/g EN/ISO 6888-1 or 2

Regulatory ComplianceISO standards and compliance

19

Food categoryMicro-

organisms

Sampling plan (20) Limits (21)

Analytical reference method (22)

Stage where the criterion

applies

Action in case of unsatisfactory

resultsm Mn c

2.2.5. Unripened soft cheeses (fresh cheeses) made from milk or whey that has undergone pasteuri-zation or a stronger heat treatment (35)

Coagulase-positive

staphylococci 5 2 10 cfu/g 100 cfu/g EN/ISO 6888-1 or 2

End of the manufacturing

process

Improvements in production hygiene. If values > 105 cfu/g are detected, the cheese

batch has to be tested for staphylococcal

enterotoxins.

2.2.6. Butter and cream made from raw milk or milk that has undergone a lower heat treatment than pasteurization

E. coli (33) 5 2 10 cfu/g 100 cfu/g ISO 16649-1 or 2End of the

manufacturing process

Improvements in production hygiene and

selection of raw materials

2.2.7. Milk powder and whey powder (32)

Entero- bacteriaceae 5 0 10 cfu/g ISO 21528-1

End of the manufacturing

process

Check on the efficiency of heat treatment and prevention of recontamination

Coagulase-positive

staphylococci5 2 10 cfu/g 100 cfu/g EN/ISO 6888-1 or 2

End of the manufacturing

process

Improvements in production hygiene. If values > 105 cfu/g

are detected, the batch has to be tested

for staphylococcal enterotoxins.

2.2.8. Ice cream (36) and frozen dairy desserts

Entero- bacteriaceae 5 2 10 cfu/g 100 cfu/g ISO 21528-2

End of the manufacturing

process

Improvements in production hygiene

2.2.9. Dried infant formulae and dried dietary foods for special medical purposes intended for infants below six months of age

Entero- bacteriaceae 10 0 Absence in 10 g ISO 21528-1

End of the manufacturing

process

Improvements in production hygiene to

minimise contamination. If Enterobacteriaceae are detected in any of the sample units, the batch has to be tested for Cronobacter and

Salmonella

1.11. Cheeses, butter and cream made from raw milk or milk that has undergone a lower heat treatment than pasteurization (10)

Salmonella 5 0 Absence in 25 g EN/ISO 6579Products placed on the

market during their shelf-life

1.12. Milk powder and whey powder (10) Salmonella 5 0 Absence in 25 g EN/ISO 6579

Products placed on the market during their

shelf-life

1.13. Ice cream (11), excluding products where the manufac-turing process or the composition of the product will eliminate the salmonella risk

Salmonella 5 0 Absence in 25 g EN/ISO 6579Products placed on the

market during their shelf-life

1.21. Cheeses, milk powder and whey powder, as referred to in the coagulase-positive staphylococci criteria in Chapter 2.2 of this Annex

Staphylo- coccal

enterotoxins5 0 Not detected in 25 g

European screening

method of the CRL for Milk

(13)

Products placed on the market during their

shelf-life

1.22. Dried infant formulae and dried dietary foods for special medical purposes intended for infants below six months of age, as referred to in the Enterobacteriaceae criterion in Chapter 2.2 of this Annex

Salmonella 30 0 Absence in 25 g EN/ISO 6579

Products placed on the market during their

shelf-life

1.23. Dried infant formulae and dried dietary foods for special medical purposes intended for infants below six months of age, as referred to in the Enterobacteriaceae criterion in Chapter 2.2 of this Annex

Cronobacter 30 0 Absence in 10 g ISO/DTS 22964

Products placed on the market during their

shelf-life

Our dairy testing products are AOAC-OMA, AOAC-PTM, and Health Canada approved.

Product AOAC-PTM AOAC-OMAMicroVal,

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FDA-BAM and AOAC standardsThe Bacteriological Analytical Manual Chapter 29 Cronobacter published by the FDA-BAM specifies both a real-time PCR method for rapid screening and a cultural method for the detection/isolation of Cronobacter spp. (3). Chromogenic agars are used to isolate the culture for confirmation. A pre-enrichment step is used to grow the bacteria to an amount (≥ 103 CFU/ mL) detectable by PCR and chromogenic agars. The cultural portion of

this method is a complete detection/isolation method, it can be used as a stand-alone method if PCR technology is unavailable. The cultural method is completely different from the ISO method. The PCR portion of the method is a screening method, whose positive results should always be confirmed with the cultural method. The PCR method may be used to confirm pure cultures as Cronobacter spp.

21

GB StandardsIn China, GB standards need to be followed: GB 4789.40-2016 is the national food safety standard for food examination of Cronobacter while GB 4789.4-2016 is for examination of Salmonella. Both standards were issued in 2016 by the China Food and Drug Administration. For Salmonella, the method is different from both ISO and FDA-BAM in the usage of culture media. For Cronobacter spp., the current GB method is very similar to the previous ISO TS 22964: 2006. Both GB standards do not describe a rapid method for testing of these pathogens.

Dairy food, according to different product categories, have their corresponding microbiological criteria in terms of testing parameters, detection limits, sampling plans & detection methods in the Chinese national

standards. GB methods are the most commonly used foodborne testing methods in which the GB 4789 series is mainly used for Microbiological testing. The GB 4789 series is based on traditional methods by adapting culture-media based, plate count methods for both routine and pathogen tests. Some rapid and instrument-based methods are used and available in China, but the relative volume of tests performed is far lower than the traditional GB methods for release testing. There has been a tendency for increased rapid method usage especially in environmental as well as in process testing in China while for some specific dairy products, rapid methods can be used as an alternative to traditional methods by enterprise guidelines.

Trends of China Food Safety Market

In recent years, the public concerns about Food Safety has been rapidly raised in China, Following the 2008 melamine event, where 50,000 infants and children were hospitalized and 6 died because the chemical melamine was added to milk and infant formula, the first “Food Safety Law” was published. Subsequent revisions were published including the latest “Food Safety Law” enacted on October 1, 2015. The Chinese government has been putting a lot of effort into improving food safety for each five-year government plan and encouraging the growth of third-party labs to improve food safety in China.

In February 2017, China issued the “13th Five-Year Plan for National Food Safety.” The “13th Five” Food Safety Plan sets out specific goals for development of China’s food safety infrastructure by 2020. The plan sets as goals, among others, to enhance sample testing to cover all types of food, develop effective governance of resource contamination, reinforce on-site inspections: establish a professional inspection team and standardize enforcement procedures and documentation, and align Chinese food safety standards with international standards.

22

Chinese GB standards for microbial dairy testing

Food category Microorganisms

Sampling plan Limits (CFU/g)

Analytical reference methodm Mn c

Cheese / Processed Cheese

Total Coliforms 5 2 100 1000 GB 4789.3 Pour Plate

S. aureus 5 2 100 1000 GB 4789.10 Pour Plate

Salmonella 5 0 0/25g -- GB 4789.4

L. monocytogenes 5 0 0/25g -- GB 4789.30

Yeast =< 50GB 4789.15

Mold =< 50

Infant formula

TVC 5 2 1000 10000 GB 4789.2

Total Coliforms 5 2 10 100 GB 4789.3 Pour Plate

Staph. aureus 5 2 10 100 GB 4789.10 Pour Plate

Cronobacter sakazakii 3 0 0/100g -- GB 4789.40 enumeration

Salmonella 5 0 0/25g -- GB 4789.4

Older infants and young children formula/ Cereal-based complementary foods for infants and young children

TVC 5 2 1000 10000 GB 4789.2

Total Coliforms 5 2 10 100 GB 4789.3 Pour Plate

Salmonella 5 0 0/25g -- GB 4789.4

Raw milk TVC =< 2E6 GB 4789.2

Pasteurized milk/Modified milk

TVC 5 2 50000 100000 GB 4789.2

Total Coliforms 5 2 1 5 GB 4789.3 Pour Plate

Staph. aureus 5 0 0/25g -- GB 4789.10 Detection

Salmonella 5 0 0/25g -- GB 4789.4

Sterilized milk Commercial Sterility GB/T 4789.26

Fermented milk

Total Coliforms 5 2 1 5 GB 4789.3 Pour Plate

Staph. aureus 5 0 0/25g -- GB 4789.10 Detection

Salmonella 5 0 0/25g -- GB 4789.4

Yeast =< 100GB 4789.15

Mold =< 30

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Food category Microorganisms

Sampling plan Limits (CFU/g)

Analytical reference methodm Mn c

Evaporated milk sweetened condensed milk and formulated condensed milk

TVC 5 2 30000 100000 GB 4789.2

Total Coliforms 5 1 10 100 GB 4789.3 Pour Plate

Staph. aureus 5 0 0/25g -- GB 4789.10 Detection

Salmonella 5 0 0/25g -- GB 4789.4

Milk powder

TVC 5 2 50000 200000 GB 4789.2

Total Coliforms 5 1 10 100 GB 4789.3 Pour Plate

Staph. aureus 5 2 10 100 GB 4789.10 Pour Plate

Salmonella 5 0 0/25g -- GB 4789.4

Whey powder and whey protein powder

Staph. aureus 5 2 10 100 GB 4789.10 Pour Plate

Salmonella 5 0 0/25g -- GB 4789.4

Cream, butter and anhydrous milkfat

TVC 5 2 10000 100000 GB 4789.2

Total Coliforms 5 2 10 100 GB 4789.3 Pour Plate

Staph. aureus 5 1 10 100 GB 4789.10 Pour Plate

Salmonella 5 0 0/25g -- GB 4789.4

Mold =< 90 GB 4789.15

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