broch_microbiological_testing_sm-4017-e.pdf

8/10/2019 Broch_Microbiological_Testing_SM-4017-e.pdf

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IntroductionThe consumers steadily growing requirements for the quality and the longer shelf lifeof foods and beverages must be met by the manu facturer. Quality assurance cant belimited to inspection of the final product alone, such as a bottled beverage or a prepared

food product. Instead, continuous inspection of incoming raw materials and in-processquality control tests must be performed throughout production. Microbiological andaseptic testing play a significant role in such quality assurance.

In the soft drink industry the microbiological and hygienic quality including thebiological stability of the products are important criteria for their assessment. The reaso just a few microbes are often all it takes to spoil large quantities of a beverage.

Although the explosive technological development has reduced the risk of contaminatioby spoiling microbes, the issue of shelf life has taken on new dimensions as a resultof the enormous production output possibilities of today. Quality control of bottling and

filling, in terms of chemical and, above all, biological stability, must be adapted to thisdevelopment by state-of-the-art test methods.

The requirements for a practical microbiological test method are that it permitsquantitative and reproducible detection of trace contamination and that it canbe performed efficiently and economically under routine conditions.These requirements are fulfilled optimally by the membrane filter method.

The principle of this method is based on the concentration of microorganisms fromrelatively large samples on the surface of the membrane filter, and on culturingthese microbes on a nutrient pad or an agar culture medium.


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Contents4 The Membrane Filter Method

6 Nutrient Pad Sets6 User Benefits

7 How to Use Nutrient Pad Sets

8 General Directions

9 Description and Typical GrowthEvaluation Results

9 1. Total Colony Count

11 2. E. coli and Coliforms,Enterobacteria

13 3. Faecal Bacteria

14 4. Non-faecal, Pathogenic Bacteria14 5. Yeasts and Molds

16 6. Product-spoiling Microorganisms

19 Troubleshooting Guide

19 Membrane Filters for Use onAgar Plates or on Aborbent Pads

21 Membrane Filters for Use withMicrosart e.motion Dispenser

22 Typical Application Examples

23 Growth comparison

24 Accessories

28 Technical Data and Application GuNutrient Pad Sets

32 Test Strains

34 Reference Guide

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Standard MF methodThe membrane filter is rinsedand then placed on a culturemedium a, b, or c and incubated.

Membrane Filter MethodThe test sample is filtered througha membrane filter

Direct MethodThe test sample is pipetted intoa petri dish

then mixed with the culturemedium and incubated

Monitor MF method

The nutrient media is given fromthe top after filtration. After a shortvacuum (


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User Benefits

Sartorius Stedim Biotech Nutrient PadSets have been used successfully in themembrane filter method for over 20 years.Practical and easy to handle, they reducelabor and simplify many microbiologicaltesting procedures.

Nutrient pads are sterile, dehydratedculturemedia. Once they are moistened with3.03.5 ml of sterile and demineralized(or distilled) water they are ready to useimmediately.

The level of moisture is optimal when anexcess ring of water surrounding the padis visible.

All Nutrient Pad Set types are suppliedwith the appropriate membranefilters, which are also presterilized andindividually packaged or dispenser-readypackaged on a band for the use withthe Microsart e.motion dispenser.

The membrane filters tailored to meet thespecial requirements of microbial detectionare available with 47 mm or 50 mmdiameters.

Nutrient pad sets (NPS) are continuouslyenhanced as part of our development pro-gram to adapt our products to changingapplication requirements. Besides thenew NPS types, we have also updated ourpackaging design. The standard NPS boxcontains 100 sterile nutrient pads, each ofwhich is individually inserted in a petridish and sterilized. Ten each of these petridishes are sealed in an aluminum bag.This special packaging in bags protectsthe sensitive formula constituents of thenutrient pads during transport and storagefrom fluctuations in humidity andtemperature. As a result, it guaranteesthe high quality of our NPS throughouttheir entire shelf life of up to 24 months.

Nutrient Pad Sets

EconomicalEliminates time-consuming and labor-intensivepreparation of culture media (sterilization and cleaning, among others).

After wetting with 3.5 mldestilled water NPS are readyto use: NPS and go

Simple to UseNutrient Pad Sets can also be used in laboratorieswhich do not have extensive microbiologicalequipment. Sterile water for moistening the pads canbe added easily with a Sartorius Stedim BiotechDosing Syringe and an attached Syringe Filter Holder(0.2 m) or with an ampoule with sterile water.

Everyone can use NPS

Consistently QualityDuring manufacture, each type of Nutrient

Pad Set is compared with the correspondingagar medium with respect to their growth- promoting properties. This QA procedure ensuresconsistent quality and reproducible results.

NPS are validated. In com-

parison to agar which is donewithin different deviationsof amount and height NPSalways give constant results

Trouble-free StorageNutrient Pad Sets have a shelf life ofup to 24 months at room temperature.

No waste or overproductionof prepared agar media

Highly VersatileNutrient Pad Sets can be modified by additives in thesolution used to wet them; for example, Wort orOrange Serum Nutrient Pads when wetted with 5 8%ethanol promote the growth of acetic-acid bacteria

Advanced system

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The forceps should alwaysbe stored in a small glasswith alcohol.

Take it out of it and flame it.

Let it cool off for a fewseconds before use.

Let the Microsart e.motionDispenser release the mem-brane filter automatically byapproaching tweezers or bypress button.

Alternatively you peel backthe transparent plastic layerof the membrane filter pack-aging manually.

Use tweezers to remove thecontent out of the packaging.

The membrane filter is placedby the tweezers onto the filtersupport of the filter holder.

The protective paper or gridshould face upwards. If thereis a protective disc make sureto discard it before assem-bling the funnel or the toppart of the filter holder.

Place the funnel on thefilter support and close it

with the clamp.For long time filtrationcover the funnel withthe lid.

Open the valve andfilter the sample.(6 oclock1)Rinse with a few mlsof sterile water toremove all productresidues or inhibitorsthat might be con-tained in the sample.

Close the valveagain.(9 oclock2).

Remove the funnel and takethe filter with the sterileforceps.

Its so easy to use Nutrient Pad Sets: NPS and go

1 Before starting with the tests remove everythingthat is not essentially needed for this work.

2 Carefully clean and disinfect your working area.3 For simple microbiological tests a laminar flowbox is not needed. When used unprofessionally, alaminar flow box increases the risk of secondarycontamination instead of protecting from it. Agood protection against airborne contamination,however, is to work close to the flame of a Bunsenburner.Instruments like forceps should be placedinto a glass with alcohol.

Label the needed amountof Nutrient Pads.

Wet the Nutrient Pad with3.5 ml of sterile, deionizedor distilled water.

Use a dosing syringe with aMinisart or a sterile pipette.

Open the lid of the Petridish only slightly to avoidairborne contamination.

Open the vacuumvalve.

(6 oclock1)Carefully flame the

filter support for~10 sec.

Close the vacuumvalve again.(9 oclock2)

Take the funnel at bothsides of the clamp andflame it from its lower sidefor ~10 sec.

Then place it on the filtersupport.

Open the vacuumvalve again.(6 oclock1)Flame the inside ofthe funnel.

Close the valveagain.(9 oclock2)

To cool it off faster,rinse with a few mlof sterile water.

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How to Use Nutrient Pad Sets

Place the filter on theNutrient Pad, avoid toentrap air bubbles under

the filter.Open the lid of the Petridish only slightly to avoidairborne contamination.

Place the Petri dish into theincubator, lid above.

Incubate strictly accordingto the recommendations.

Evaluate immediately afterthe end of theincubation time.

3 oclock After the Filtration RunThe residual vacuum between thepump and valve is released via theventing filter.

12 oclock For AutoclavingFor reliable sterilization, the steam flowsfreely through all openings.

6 oclock For FiltrationThe tap is open. The full vacuum iseffective at the filter support| membranefilter. The venting filter is off-line.

9 oclock After FiltrationThe tap is closed. The vacuum betweenthe valve and membrane filter is releasedunder sterile conditions. Secondarycontamination of the bottom of thefilter is ruled out entirely.

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2)

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Do not touch bacterial matter withyour hands.

Before and after use, inoculating loops andwires must be sterilized by flaming untilthey glow red-hot.

All laboratory equipment which has comein contact with bacteria must be sterilized.

To protect people and animals fromcontagious diseases or poisoning, livingcultures have to be destroyed beforecleansing or disposing of the containers.One method is to coat them thoroughlywith disinfectants or to autoclave themin suitable containers.

Sartorius Stedim Biotech Nutrient Pads areparticipating regularly at official inter-lab-oratory tests for the microbiological inves-tigation of drinking water according to theNew European Drinking Water Guideline.This certificate of the NiederschsischenLandesgesundheitsamt in Aurich (publichealth agency, Lower Saxony) quotea reference for the passed tests with goodsuccess.

General ProcedureTo obtain reliable results for microbio-logical tests, it is necessary to work underconditions that rule out contamination bymicroorganisms which distort such results.

That is why it is recommended to work nearthe flame of a Bunsen burner in a roomprotected from drafts. Before beginningwith the actual procedure, spraying orwashing down the working area with a dis-infectant is mandatory (e.g., 70% alcohol).

Before use, filter holders, tweezers andscissors should be sterilized by one ofthe standard methods, such as flamingfor routine tests.

How to Handle MicroorganismsMicroorganism cultures must always behandled as carefully as if they containedpathogens.

Working with microorganisms is notdangerous if the following safety rulesare observed:

Wash your hands thoroughly before andafter working in a laboratory.

Do not eat or drink in a laboratory.

General Directions

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Bacillus subtilis

Mixed culture from well water

Staphylococcus aureus

Mixed culture from process water

Description and Typical Growth Evaluation Results1. Total Colony Count

Standard TTC (I mod.) NPSType 14055

Meat extract-peptone medium for deter-mining the total CfU count based onthe APHA (water) and modified by theaddition of TTC.Dehydrated culture medium for cultivatimicroorganisms in raw materials, water(general quality), waste water, beveragesbeer, foods and other products.

References:APHA (water), ISO 7704, VLB

Incubation Conditions:< 5 days at 3035C

Evaluation and Typical Results:Predominantly bacteria grow on thismedium. The majority of their coloniesare stained red by TTC reduction.

Caso NPSType 14063

Soybean-Casein Digest medium for isolatingmicroorganisms and for determining thetotal CfU count. Dehydrated culturemedium for cultivating microorganisms inpharmaceuticals, cosmetics, raw materials,water (general quality), waste water, foodsand other products.

References:APHA (dairy), APHA (food), APHA (water),AOAC, DAB, EG 98/83, EP, FDA, IDF, ISO 7704,ISO 8199, ISO 9308-1 [1990], ISO 9308-1[2001], USDA, USP

Incubation Conditions:Bacteria:< 3 days at 3035C Yeasts and molds:< 5 days at 3035C

Evaluation and Typical Results:Predominantly bacteria of different sizes,shapes and colors. Remarks: Depending on themicrobes to be detected, this medium can beconverted into a selective one. When 10%serum is added to the wetting liquid a numberof fastidious pathogenic bacteria like the gen-era Pneumococcus, Neisseria, Streptococcus,Corynebacterium, Erysipelothrix and Brucellaare able to grow on the medium.

R2A NPSType 14084

Low nutrient medium for the enumerationof heterophilic organisms in treatedpotable water and highly purified water.Growth medium for microorganismswhich have adapted to the particular livingconditions of water low in nutrients.Dehydrated culture medium for cultivatingmicroorganisms in water for pharmaceuti-cal purpose, water (general quality), wastewater and other products.

References:APHA (water), EP, ISO 7704

Incubation Conditions:> 5 days at 3035C

Evaluation and Typical Results:Predominantly bacteria grow on this medi-um. Their colonies are of different size andcolor, most of them are white or colorless.Remarks: Stressed and chlorine-tolerantbacteria are stimulated by this mediumin combination with lower incubationtemperatures and longer incubation time.

Escherichia coli

Mixed culture from water

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2. E. Coli and Coliforms, Enterobacteria

Escherichia coli

Mixed culture from water

CHROMOCULT* NPSType 14087

For the detection of total coliforms andEscherichia coli. Dehydrated culturemedium for cultivating microorganismsin raw materials, water (general quality),waste water, beverages, foods andother products.

References:ISO 7704, Journal Food Protection,ZenHyg (journal of hygiene)

Incubation Conditions:2028 hours at 36 2C

Evaluation and Typical Results:E. coli develops dark-blue to violet colonies,other coliforms red to pink colonies. Othergram-negative colonies are colorless, a fewwith -Glucuronidase activity are light blueto turquoise. Remarks: To confirm E. coligive one drop of Kovacs indole reagent oneach dark blue colony. Cherry red colorafter a few seconds is a positive reaction.

* Trade mark owner and manufactureris Merck KGaA

Endo NPSType 14053

Selective medium for detecting andenumerating E. coli and coliform bacteriDehydrated culture medium for cultivatimicroorganisms in raw materials, water(general quality), natural water, wastewater, beverages, soft drinks, concentratfruit juice, sugar, sugar products, foodsand other products.

References:APHA (dairy), APHA (food), APHA (wDGHM, ISO 7704, ISO 9308-1 [1990], MUSDA


Evaluation and Typical Results:E. coli form red colonies with a metallicsheen and a red dot at the underside ofthe membrane. Other coliforms grow asdark to light red colonies without metallisheen. Colorless colonies of lactose-negative bacteria are not counted.

Escherichia coli

E. coli and coliforms from river water

ECD NPSType 14082

Selective culture medium for detectingand identifying Escherichia coli. Bile saltinhibits the accompanying flora of microbesnot living in the intestine. Dehydratedculture medium for cultivating micro-organisms in raw materials, water(general quality), waste water, beverages,foods and other products.

References:APHA (water), DIN 10110, EG 98/83,ISO 7704, ISO 8199, ISO 9308-1 [2001],LMBG, USDA

Incubation Conditions:1618 hours at 442C

Evaluation and Typical Results:Colonies that show light blue fluorescenceunder UV light indicate E. coli; confirma-tion with a drop of KOVCS indole reagentis required, a positive reaction is shownby a cherry color after a few seconds.Remarks: This medium can be used for therapid detection of Escherichia coli basedon the ISO 9308-1.

Escherichia coli

E. coli colonies fluoresence in UV light

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Escherichia coli

E. coli and coliforms from waste water

Teepol NPSType 14067

Lauryl Sulphate medium for the detectionof E. coli and faecal coliform bacteriaaccording to Burman, N.P. (1967).Dehydrated culture medium for cultivatingmicroorganisms in water (general quality),waste water, beverages, foods and otherproducts.

References:AFNOR, APHA (water), BS, FDA, ISO 7704,ISO 9308-1 [1990], USDA


Evaluation and Typical Results:E. coli and coliform bacteria form 12 mmdiameter yellow colonies surroundedby a yellow zone. Non-lactose fermentingbacteria develop red or colorless colonieswithout yellow zone.

Escherichia coli

E. coli and coliforms from river water

MacConkey NPSType 14097

For the isolation and differentiation of coli-form bacteria and other enterobacteriaceae.Dehydrated culture medium for cultivatingmicroorganisms in pharmaceuticals,cosmetics, raw materials, water (generalquality), natural water, waste water,beverages, soft drinks, concentrates, fruit juice, foods and other products.

References:APHA (dairy), APHA (food), APHA (water),AOAC, DAB, DIN 38411, DGHM, EP, ISO7704, LMBG, MNO, USDA, USP


Evaluation and Typical Results:Escherichia coli forms large red or reddishcolonies, coliform microbes form large pink,sometimes slimy colonies, lactose-negativeenterobacteria form colorless colonies.Gram-positive microbes are inhibited.

m FC NPSType 14068

For the detection of E. coli and faecal coli-form bacteria according to Geldreich et al.Dehydrated culture medium for cultivatingmicroorganisms in raw materials, water(general quality), waste water, beverages,foods and other products.

References:APHA (food), APHA (water), AOAC, EPA,FDA, ISO 7704, ISO 9308-1 [1990], USDA


Evaluation and Typical Results:E. coli and coliform bacteria form bluecolonies with a blue surrounding. This coloris dark blue at faecal coliforms with stronglactose fermentation and lighter blue fornon-faecal coliforms with weaker lactosefermentation. Lactose-negative bacteriagrow with different colors and are notevaluated. Remarks: Higher incubationtemperatures largely suppress the non-faecal coliforms.

Escherichia coli


2. E. Coli and Coliforms, Enterobacteria

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Tergitol TTC NPSType 14056

Selective and differential medium forthe detection and enumeration of coliformbacteria and E. coli according to Pollard;modified acc. to Chapman. Dehydratedculture medium for cultivating micro-organisms in raw materials, water (generalquality), waste water, beverages, foodsand other products.

References:APHA (food), EG 98/83, ISO 7704, ISO 8199,ISO 9308-1 [1990], ISO 9308-1 [2001]

Incubation Conditions:

1824 hours at 36 2CEvaluation and Typical Results:Lactose-positive microorganisms formyellow-orange colonies with a yellowsurrounding and have a yellow dot underthe membrane filter. According toISO 9308-1 all colonies that show yellowcolor under the membrane filter arecounted as positive. Remarks: Tergitol 7inhibits Gram positive colonies and mini-mizes the swarming of Proteus. Furtherdifferentiations of E.coli and coliforms

with Oxidase- and Indol-Tests are required.

Escherichia coli


3. Other Faecal Bacteria

Azide NPSType 14051

For the detection and enumeration ofintestinal enterococci according to Slanetzand Bartley. Dehydrated culture mediumfor cultivating microorganisms in rawmaterials, water (general quality), naturalwater, waste water, beverages, foods andother products.

References:APHA (food), APHA (water), EG 98/83,HMSO, ISO 7704, ISO 7899-2, ISO 8199,LMBG, MNO


4048 hours at 362CEvaluation and Typical Results:Enterococci form red, pink or reddishbrown colonies with a diameter of0.52 mm. Remarks: Enterococci areconsidered to be indicator organismsof faecal contamination. They are lesssensitive to chemical effects than areE. coli organisms and are therefore longerdetectable, for instance in waste waterand in chlorinated water.

Enterococcus faecalis

Enterococci from waste water

Salmonella typhosa, streak

Salmonellae from waste water

Bismuth Sulfite NPSType 14057

Selective culture medium according toWilson and Blair for isolating Salmonelltyphi and other salmonellae. Dehydratedculture medium for cultivating microorgisms in pharmaceuticals, cosmetics, rawmaterials, water (general quality), wastewater, foods and other products.

References:AFNOR, APHA (dairy), APHA (food), ADGHM, FDA, HMSO, ISO 6579 [1981]ISO 7704, USDA, USP


4048 hours at 36 2CEvaluation and Typical Results:Most salmonellae form light colored colowith brown to black centers surrounded bblack zone with a metallic sheen (fish eySome Salmonella species develop uniformdark brown to black colonies which may the typical zone. Remarks: If a very slighcontamination with salmonellae is suspecprepare a selective enrichment culture ansubsequently streak the sample with aninoculation loop on a membrane filter tha

has been placed on the pre-wetted NPS.

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4. Non-faecal, Pathogenic Bacteria

Pseudomonas aeruginosa

Mixed culture with Pseudomonas aeruginosa

Cetrimide NPSType 14075

For the detection and enumeration ofPseudomonas aeruginosa according toLowbury. Dehydrated culture medium forcultivating microorganisms in cosmetics,raw materials, water (general quality),waste water, foods and other products.

References:APHA (water), AOAC, ASM, DIN 38411,EG 98/83, FDA, ISO 7704, ISO 8199,EN 12780, EN ISO 16266


Evaluation and Typical Results:Pseudomonas aeruginosa forms blue,blue-green or yellow-green colonieswith 12 mm diameter and blue zones.The colonies produce pyocyanin andfluorescein and show fluorescence inUV-light. Other Pseudomonads developcolonies with different colors. Remarks:Further tests are necessary for definitiveidentification of Ps. aeruginosa.

Staphylococcus aureus

Mixed culture of staphylococci

Chapman NPSType 14074

Mannitol salt medium according toChapman, modified for detecting andisolating pathogenic Staphylococci.Dehydrated culture medium for cultivatingmicroorganisms in pharmaceuticals,cosmetics, raw materials, water (generalquality), waste water, foods and otherproducts.

References:APHA (food), AOAC, DGHM, FDA, HMSO,ISO 7704, USP


Evaluation and Typical Results:Staphylococcus aureus forms yellowcolonies with a yellow surrounding(mannitol-positive). Other Staphylococcigrow without zones of color change.Most other bacteria are inhibited.

5. Yeasts and Molds

Lysine NPSType 14061

Selective medium for isolating andenumerating wild yeasts in breweries acc.to Morris and Eddy. Dehydrated culturemedium for cultivating microorganisms inbeer and other products.

References:Journal Institute of Brewing, VLB

Incubation Conditions:35 days at 3035C

Evaluation and Typical Results:Only wild yeasts (not belonging to thegenus Saccharomyces) which utilizelysine as sole source of nitrogen grow onthis medium, they form white or creamcolored colonies; brewery culture yeastsgrow not at all or very poorly.

Torulopsis spec.

Wild yeasts from lager beer

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6. Product-spoilingMicroorganisms

Glucose Tryptone NPSType 14066

For the enumeration of mesophilic andthermophilic bacteria, especially flat-sourmicroorganisms in canned foods.

References:APHA (dairy), APHA (food), AOAC, ICUMSIFU, ISO 7704, NCA

Incubation Conditions:1872 hours at 3035C for mesophilicbacteria; 4872 hours at 552C forthermophilic sporulating microorganisms

Evaluation and Typical Results:Microorganisms that ferment glucoseand produce acid grow as yellowish greencolonies. Flat-sour colonies have adiameter of 2-5 mm, a yellowish-greencolor and are surrounded by a yellow zone.Remarks: For the incubation at 552C thepetri dishes must be placed into a moistchamber.

Bacillus coagulans, the flat sour colony

Mixed culture from canned vegetables

Wort NPSType 14058, 14092

For the detection and determination ofyeasts and molds. Dehydrated culturemedium for cultivating microorganismsin raw materials, beverages, beer, wine,soft drinks, concentrates, foods and otherproducts.

References: VLB

Incubation Conditions:35 days at 2025C or at 3035Cdepending on the target of theinvestigation

Evaluation and Typical Results: Yeasts usually develop smooth white orcolored colonies. Molds generally formvelvety or fluffy cotton-like colonies thatlook white in the early growth phase andmay take various colors after conidiosporeproduction.

Saccharomyces cerevisiae

Yeasts and molds from spoiled beer

Wallerstein (WL Nutrient) NPSType 14089

For the detection and enumeration of the

microbiological flora of brewing and fer-mentation processes acc. to Green and Gray(1950). Dehydrated culture medium forcultivating microorganisms in beverages,beer, wine, soft drinks, concentrates, fruit

juice and other products.

References:ISO 7704

Incubation Conditions:25 days at 3035C aerobic or anaerobicdepending on the target of the investigation

Evaluation and Typical Results: Yeasts usually grow as yellowish greencolonies. Molds generally form velvety orfluffy cotton-like colonies that look whitein the early growth phase and may takevarious colors after conidiospore produc-tion. Bacteria grow slowly and their coloniesare of different size and color. Remarks:The addition of 0.004 g/l cycloheximide tothe wetting solution makes the mediumselective for lactic acid bacteria, the growthof yeasts and molds is suppressed.

Saccharomyces cerevisiae

Lactobacillus plantarum

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Jus de Tomate (Tomato Juice) NPSType 14079

For the detection of product spoiling lacticacid bacteria especially Oenococcus oeniacc. to Dubois, Bindan and Lafon-Lafour-cade. Tight-fitting, special petri dishes formicroaerophilic incubation. Dehydrated cul-ture medium for cultivating microorganismsin wine, fruit juice and other products.

References:ISO 7704, Lanaridris& Lafon-Lafourcade

Incubation Conditions:57 days at 3035C anaerobic(microaerophil); control for slowlygrowing micro-organisms after10 days is recommended

Evaluation and Typical Results:Lactobacilli form compact, whitish toslightly yellowish colonies with 13 mmdiameter. Pediococci develop somewhatsmaller colonies with approx. 1 mm dia-meter that later get a whitish to slightlybrownish color. Oenococcus oeni growsas colorless to whitish colonies with adiameter smaller than 1 mm. Remarks:This medium must be incubated underanaerobic to microaerophilic conditions.

Lactic-acid bacteria, streak

Oenococcus oeni from wine


Orange Serum NPSType 14062; 14096

For the isolation and enumeration ofacid-tolerant microorganisms. Dehydratedculture medium for cultivating micro-organisms in raw materials, water (generalquality), waste water, wine, soft drinks,concentrates, fruit juice, foods and otherproducts.

References:APHA (water), IFU, ISO 7704, MPP(packaging staff)

Incubation Conditions:35 days at 3035C aerobic oranaerobic depending on the target ofthe investigation

Evaluation and Typical Results:Only acid-tolerant microorganisms cangrow on this medium such as lactic acidbacteria (Lactobacillus, Pediococcus etc.),acetic acid bacteria, yeasts and molds.Remarks: This medium is available withpH 5.5 and with pH 3.2.

Rhodotorula spec.

Mixed culture from a soft drink

VLB-S7-S NPSType 14059

For the detection of pediococci and lactobacilli according to Emeis; modified accto Rinck and Wackerbauer. Dehydratedculture medium for cultivating micro-organisms in beer and other products.

References:EBC, ISO 7704, MEBAC, VLB

Incubation Conditions:35 days at 3035C anaerobic(microaerophil)

Evaluation and Typical Results:Pediococci (Sarcina) develop round pagreen colonies with smooth peripheriesand approx. 1 mm in diameter. Lactobacgrow as slightly rounded, irregularly lobcolonies with approx. 2 mm in diameterwhich are initially light green and laterdark green. Remarks: This medium mustbe incubated under anaerobic to micro-aerophilic conditions.

Lactobacillus brevis

Lactobacilli and pediococci from sediment, streak

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Weman NPSType 14065

For the detection and determination ofslime-forming mesophilic bacteria accord-ing to Weman, modified acc. to Lorenz.Dehydrated culture medium for cultivatingmicroorganisms in soft drinks, concen-trates, sugar, sugar products and otherproducts.

References:ICUMSA, ISO 7704

Incubation Conditions:35 days at 3035C

Evaluation and Typical Results:The colonies of slime-forming mesophilicbacteria are smooth, round, usuallycolorless and transparent or translucent.Some have a diameter greater than 5 mm.

MRS NPSType 14077

For the detection of different Lactobacillusspecies and other lactic acid bacteriaaccording to DE MAN, ROGOSA andSHARPE. Dehydrated culture mediumfor the isolation and cultivation of Lacto-bacillus from dairy and food products.

References:APHA (food, dairy products)

Incubation Conditions:35 days at 30C anaerobic conditions

Evaluation and Typical Results:The MRS NPS are used for the detectionof a variety of Lactobacilli. The Lactobacillispecies grow as slightly rounded whitishcolonies with approximately 12 mm indiameter. Other microorganisms, whichhave not this typical growth, can bedefined by confirmation tests. Lactobacillispecies are gram positive, Katalase negativeand negative in the production of Indoleand Hydrogen sulfide.

Leuconostoc mesenteroides

Mixed culture from sugar syrup

Lactobacillus brevis

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Troubleshooting GuideIf the directions for use are not beeingfollowed this may lead to unsatisfactoryresults listed below:

1. Inhibited Growth, Tiny Colonies pad too dry: not enough water used

2. Colonies Run

pad too wet, water film on themembrane filter: too much water used.

Colonies of motile microbes (such asBacillus or Proteus) tend to run eventhough the water dosage is correct. Toprevent this, add NaCI or an emulsifier.

3. Contamination from Underneath Inhibited colony growth, excess ring

of liquid cloudy, often includingdiscoloration of the pad:

membrane placed with grid facedownon the pad instead of faceup

contamination during rehydration(by airborne microbes, by contact orby contaminated water)

If agar plates or absorbent pads to bewetted with liquid culture medium areused instead of Nutrient Pad Sets, werecommend Sartorius Stedim Biotechcellulose nitrate (cellulose ester) mem-

brane filters. These membranes areoffered in a choice of three differentcolors to suit your specific test applica-tion, and provide a high-contrastbackground.For simple evaluation ofthe results, a grid divides the filtrationarea into 130 squares, each measuring3.1 + 3.1 mm.

Membrane Filters for Use onAgar Plates or on Adsorbent Pads

contamination during preparation

microbes rinsed off the membrane filby incomplete vacuum filtration ofthe sample or rinse liquid or by tiltingthe prepared petri dish

contaminated filter support

contaminated forceps

4. Growth on One Side Only

petri dish slanted in the incubator

5. Too Profuse or too Sparse Growth(optimum microbial number between20 and 200 per filter)

wrong dilution selected or sampleinadequately mixed with the diluent.

6. Non-uniform Growth

sample volume less than 5 ml filteredwithout adding sterile NaCl-buffer-solution as a diluent or sample voluminadequately mixed with the diluent.

The membrane filters are available indiually packaged and sterilized or packagin a special designed individual packagon a band for the use with the Microsar e.motion membrane filter dispenser.

The certificate included in every packagdocuments the quality assurance testsas well as the compliance of the 0.45 mmembrane filters with ISO 7704.

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White Membrane with Black Grid

Pore Size d Pckg. Size Order No.

0.2 m 47 100 11407-47-ACN*47 1,000 11407-47-ACR*50 100 11407-50-ACN*50 1,000 11407-50-ACR

0.45 m 47 100 11406-47-ACN*47 1,000 11406-47-ACR*50 100 11406-50-ACN*50 1,000 11406-50-ACR*

HighFlow 0.45 m 47 100 114H6-47-ACN

47 1,000 114H6-47-ACR50 100 114H6-50-ACN50 1,000 114H6-50-ACR

0.65 m 47 100 11405-47-ACN*50 100 11405-50-ACN

0.8 m 47 100 11404-47-ACN*47 1,000 11404-47-ACR50 100 11404-50-ACN*

1.2 m 47 100 11403-47ACN*47 1,000 11403-47ACR50 100 11403-50ACN*50 1,000 11403-50ACR

White Membrane with Green Grid

Pore Size d Pckg. Size Order No.0.45 m 47 100 13906-47-ACN*

47 1,000 13906-47-ACR*50 100 13906-50-ACN*50 1,000 13906-50-ACR*

HighFlow 0.45 m 47 100 139H6-47-ACN

47 1,000 139H6-47-ACR50 100 139H6-50-ACN

0.65 m 47 100 13905-47-ACN1.2 m 47 100 13903-47-ACNOther types on request

Green Membrane with Dark Green Grid


0.45 m 47 100 13806-47-ACN*47 1,000 13806-47-ACR*50 100 13806-50-ACN*50 1,000 13806-50-ACR*

Grey Membrane with White Grid


0.45 m 47 100 13006-47-ACN*47 1,000 13006-47-ACR*50 100 13006-50-ACN*50 1,000 13006-50-ACR

0.65 m 47 100 13005-47-ACN*50 100 13005-50-ACN*50 1,000 13005-50-ACR

0.8 m 47 100 13004-47-ACN*47 1,000 13004-47-ACR50 100 13004-50-ACN*

8 m 47 100 13001-47-N(non-sterile)

50 100 13001-50-N(non-sterile)

Prefilters, White Without Grid11301, a white membrane filter with apore size of 8 m is used as a prefilter ina special prefilter attachment (16807) forbacteriological analyses. It retains coarsesuspended particles, whereas it allowsmicroorganisms to pass through. Thesemicrobes are trapped on the surface of theunderlying bacteria-retentive membrane

filter. Order no.: 11301--47----ACN and11301--50----ACN.

* Also available as a non-sterile version.To order boxes with 100 pcs. replace ACN with Nand for boxes of 1,000 pcs. replace ACR with R.

For Detection of Bacteriain Dyed Media.

Providing Optimal Contrast to Light-colored or Transparent Bacteria Colonies.

For Detection of Yeastsand Molds.

Membrane Filters for Use on Agar Plates or on Absorbent Pads

The special pore structure of the0.45 m HighFlow membranefilters allow shorter filtrationtimes due to higher flow ratesand throughputs. Especially E. colishows best growth promotion onHighFlow membranes.As every Sartorius Stedim Biotech0.45 m membrane filter lot thesemembranes are also tested andreleased according to ISO 7704.

HighFlow

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White Membrane with Black Grid

Pore d Pckg. Order No.

Size Size0.2 m 47 3+ 100 11407Z-47----SCM

50 3+ 100 11407Z-50----SCMHighFlow 0.45 m 47 3+ 100 114H6Z-47----SCM

50 3+ 100 114H6Z-47----SCM0.45 m 47 3+ 100 11406Z-47----SCM

50 3+ 100 11406Z-47----SCM0.8 m 47 3+ 100 11404Z-47----SCM1.2 m 47 3+ 100 11403Z-47----SCM

50 3+ 100 11403Z-50----SCM

White Membrane with Green Grid

Pore d Pckg. Order No.Size Size0.45 m 47 3+ 100 13906Z-47----SCM

50 3+ 100 13906Z-50----SCMHighFlow 0.45 m 47 3+ 100 114H6Z-47----SCM

50 3+ 100 114H6Z-47----SCM

Green Filter with Dark Green Grid


Size Size0.2 m 47 3+ 100 15407Z-47----SCM0.45 m 47 3+ 100 13806Z-47----SCM

50 3+ 100 13806Z-50----SCM

Grey Filter with White Grid


Size SizeHighFlow 0.45 m 47 3+ 100 130H6Z-47----SCM0.45 m 47 3+ 100 13006Z-47----SCM

50 3+ 100 13006Z-50----SCM0.65 m 47 3+ 100 13005Z-47----SCM

50 3+ 100 13005Z-50----SCM0.8 m 47 3+ 100 13004Z-47----SCM

50 3+ 100 13004Z-50----SCM

For Detection of Bacteriain Dyed Media.

Providing Optimal Contrast to Light-colored or Transparent Bacteria Colonies.

For Detection of Yeastsand Molds.

Membrane Filters for Use with Microsart e.motion Dispenser

The special pore structure of the0.45 m HighFlow membranefilters allow shorter filtrationtimes due to higher flow ratesand throughputs. Especially E. colishows best growth promotion onHighFlow membranes.As every Sartorius Stedim Biotech0.45 m membrane filter lot thesemembranes are also tested andreleased according to ISO 7704.

HighFlow

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Typical Application ExamplesProduct Detection and Enumeration of... Nutrient Pad TypeBeer Lactobacilli and Pediococci and other beer spoiling organisms VLB-S7-S, MRS Total colony count Standard, Standard TTC Wild yeasts Lysine

Yeasts and molds Malt Extract*, Wallerstein Nutrient, WortDairy Latic Acid Bacteria | Lactobacillaceae MRSFoods Acid-tolerant microorganisms Orange Serum

Enterobacteria, E. coli and coliforms CHROMOCULT**, ECD, Endo, (MacConkey), m FC, Teepol |Lauryl Sulphate, Tergitol TTC

Enterococci, Enterococcus faecalis Azide | KF Strep Lactobacilli MRS Pseudomonas aeruginosa Cetrimide

Salmonellae Bismuth SulfiteStaphylococci, Staphylococcus aureus ChapmanThermophilic spore formers and mesophilic bacteria Glucose TryptoneTotal colony count Caso, Standard, Standard TTC, TGE | Tryptone Glucose Extract

Yeasts and molds Malt Extract, WortFood & beverage Lactobacilli MRSFruit juice Enterobacteria, E. coli and coliforms Endo, (MacConkey), Tergitol TTC* Lactobacilli MRS Oenococcus and other product spoiling organisms Jus de Tomate | Tomato Juice, Orange Serum Yeasts and molds Malt Extract, Schaufus Pottinger | m Green yeast and mold,

Wallerstein Nutrient, WortMilk E. coli and coliforms Endo Enterococci, Enterococcus faecalis Azide | KF Strep Salmonellae Bismuth SulfitePharmaceuticals, WFI, Enterobacteria, E. coli MacConkeyraw materials and cosmetics Enterococci, Enterococcus faecalis Azide | KF Strep Pseudomonas aeruginosa Cetrimide (cosmetics only) Staphylococci, Staphylococcus aureus Chapman

Total colony count Caso, R2A

Yeasts and molds, Candida albicans SabouraudSoft drinks, concentrates Acid-tolerant microorganisms, Lactic-acid bacteria Orange Serum, VLB-S-7-S Enterobacteria, E. coli and coliforms Endo, MacConkey Lactobacilli MRS Mesophilic slime-forming bacteria, Leuconostoc Weman Total colony count Standard*, Standard TTC*, TGE | Tryptone Glucose Extract Yeasts and molds Malt Extract, Schaufus Pottinger | m Green yeast and mold,

Wallerstein Nutrient, WortSugar, sugar products E. coli and coliforms Endo Mesophilic slime-forming bacteria, Leuconostoc Weman Thermophilic spore formers and mesophilic bacteria Glucose Tryptone

Yeasts and molds Malt Extract*, Schaufus Pottinger | m Green yeast and mold, Wort*Water (general quality), Acid-tolerant microorganisms, Lactic-acid bacteria Orange Serummineral water, natural water,

Enterobacteria, E. coli and coliforms CHROMOCULT**

, ECD, Endo, (MacConkey), m FC, Teepol |waste water Lauryl Sulphate, Tergitol TTC Enterococci, Enterococcus faecalis Azide | KF Strep Pseudomonas aeruginosa Cetrimide

Salmonellae Bismuth SulfiteStaphylococci, Staphylococcus aureus ChapmanTotal colony count Caso, R2A, Standard, Standard TTC, TGE | Tryptone Glucose Extract,

Yeast Extract Yeasts and molds, Candida albicans SabouraudWine Acetobacter Orange Serum, Wort (both wetted with 5-8% ethanol)

Acid-tolerant microorganisms, Lactic-acid bacteria Orange Serum Lactobacilli MRS Oenococcus and other wine spoiling organ. Jus de Tomate | Tomato Juice Yeasts and molds Malt Extract, Schaufus Pottinger | m Green yeast and mold,

Wallerstein Nutrient, Wort * These NPS types are suitable for the determination of the mentioned microorganisms, although the media are not explicit declared

in the references described in this publication. ** Trade mark owner and manufacturer is Merck KGaA22


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Growth of E. Colion Endo NPS

E. coli forms red colonies with a metallicsheen. Other coliforms would grow asdark to light red colonies without metallicsheen.

E. coli shows no metallic sheen on thismixed esters membrane. Therefore it is verydifficult to differenciate between E. coliand coliforms without any further test. Aquantitative statement is difficult due tothe fact of running colonies on the mixedesters membrane surface.

Growth of Pseudomonas Aeruginosaon Cetrimide NPS

Pseudomonas aeruginosa forms blue,blue-green or yellow-green colonies with12 mm diameter and blue zones. The colo-nies produce pyocyanin and fluoresceinand show fluorescence in UV-light. OtherPseudomonads would develop colonieswith different colors.

On this mixed esters membrane grow lesscolonies and without the blue zone. Due tothe variance in the allocation of the pores,here the mixed esters membrane did notguarantee a sufficient nutrient supply. Thismay cause in false negative results.

Mixed esters membrane Mixed esters membrane

Sartorius Stedim Biotech cellulose nitrate membraneSartorius Stedim Biotech cellulose nitrate membrane

Growth ComparisonThe principle of the membrane filtermethod is based on the concentrationof microorganisms from relatively largesamples on the surface of a membranefilter. Nutrients and metabolites areexchanged through the pore system ofthe membrane filter. The pore size aloneis not a meaningful criterion. Due to the

variance in allocation of the pores, not allmembranes guarantee sufficient nutrientsupply. A comparison of Sartorius StedimBiotech cellulose nitrate (cellulose mixedester) membranes with other mixed estermembranes reveals significant differencesin growth promotion results.

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Accessories

Combisart 3-Branch Manifold PlusBiosart 250 FunnelsThe Biosart 250 Funnel has been designedfor microbiological quality assurance inindustry. The sterile 250 ml (50 ml gradua-tions) plastic funnel guarantees fast filtra-tion and high sample throughputs duringroutine testing. Its large inner diameterallows high flow rates, and the taperedinner wall permit thorough flushing ofthe funnel, after filtration.

16407-25-ALK Biosart 250 Funnels,50 units, sterile-packaged

16407-25-ACK Biosart 250 Funnels,50 units, individuallysterile-packaged

For further infomation about ourCombisart manifolds and accessories,please consider our Combisart brochure.

Combisart 3-Branch Manifold PlusBiosart 100 MonitorsBiosart 100 Monitors are sterile disposableswith an incorporated membrane filter andcellulose pad. They are ready-to-use andafter filtration, the funnel will be removed,so the lid and the base fit to a petri dish.Each box contains 48 units with 47 mm,gridded membrane filters.

16401-47-07-ACK Biosart 100 Monitor,individually sterile-packaged, 0.2 mwhite | black grid

16401-47-06-ACK Biosart 100 Monitor,individually sterile-packaged, 0.45 mwhite | black grid

16402-47-06-ACK Biosart 100 Monitor,individually sterile-packaged, 0.45 mgreen | dark green grid

16403-47-06-ACK Biosart 100 Monitor,individually sterile-packaged, 0.45 mgrey | white grid

16414 Biosart 100 Adapter(altern. 16424)

Combi.jet Manifold plus Microsart Funnel 100 and Microsart e.jetTransfer PumpMicrosart Funnel 100 | 250 are sterile100 ml & 250 ml funnels. The optimalsealing is guaranteed by a click-fit closure.The large inner diameter ensures a highflow rate and the optimized shape allows athorough rinsing of the system subsequentto the filtration. No liquid is retained inthe funnel.

16A07--10------N Microsart Funnel 100,sterile in 5 sealed bags

16A07--25------N Microsart funnel 250,sterile in 6 sealed bags

166MP-4 Microsart e.jetTransfer pump

16848-CJ Microsart Combi.jet2-fold manifold

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Combisart Individual Systems andFilter HoldersFor low number of samples to test, theindividual system is ideal to use. In thisequipment set-up, you simply use a siliconestopper and a single base to fit your choiceof funnel type on a suction flask.

16841 Stainless steel single base6981065 Stainless steel funnel, 100 ml6981002 Stainless steel funnel, 500 ml17575-ACK Minisart SRP 25, 50 sterile

venting filters17173 Silicone stopper16672 Suction flask

Alternatively to position 13 you canuse 16219-CS as 100 ml filter holder or16201-CS as 500 ml filter holder.

Vacuum Pumps, Water Traps and Vacuum HoseThe vacuum pumps are neoprenemembrane pumps with low noise level,oil- and maintenance-free, reliable sourcesof vacuum. The water traps are preventingan overflow of filtrate into the vacuumpump.

16694-2-50-22 Microsart maxi.vac formultiple filtration runs,230 V, 50 Hz

16694-1-60-22 Microsart maxi.vac,115 V, 60 Hz

16694-2-50-06 Microsart mini.vacfor single filtration run,230 V, 50 Hz

16694-1-60-06 Microsart mini.vac,115 V, 60 Hz

17804-M Vacusart, 3 individuallysterile-packaged PTFEfilter

166MP-4 Microsart e.jet TransferPump

16610 Woulffs bottle, 500 ml,with stop cock

16623 Rubber vacuum hose, 1 m

Stainless Steel Prefilter AttachmentFor removal of coarse particulate sub-stances from samples in a single stepalong with bacteria-retentive filtrationfor subsequent microbiological testing.Clips between a filter support (16840 or16841) and a stainless steel funnel (asshow at the photo) or Biosart 250 FunnelAutoclavable and can be flamed.

16807 Prefilter attachment

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Dosing Syringe | Colony CounterThe most convenient way to moist theNPS with water is to use a dosing syringewith an adapted Minisart syringe filter.Simultaneous sterilization and dosing ofdemineralized water in 3.5 ml steps is easydone by dropping the sinker at the endof the suction tubing into the water,and the dosing syringe filled and dosedby operating the twigger automatically. Compact battery operated colony counter,is as simple to use as a ball-point pen, andhas a 4-digit LCD-display.

16685-2 Dosing syringe17597k Minisart, 0.2 m, individually

sterile-packaged17649 Colony Counter

Microsart e.motion Dispenser Membrane Filters on DemandThe completely new membrane filterdispenser meets all requirements placedon advanced laboratory equipment.The membrane filters are released fromtheir sterile packaging fully automaticallyat the touch of a button or hands-free a dispensing operation is triggered whenthe optical sensor detects approachingtweezers.

16712 Microsarte.motion Dispenser

Microsart e.motion Membrane FiltersThe cellulose nitrate (cellulose mixed ester)membranes suitable for the use in dispens-ers are sterile-sealed, without protectivepaper on top of each filter, in a speciallydesigned individual package on a band.The special pleating of the band of mem-brane filter units ensures that they areperfectly flat when dispensed. The shapeof the sealed band guarantees uniformdispensing of the individual membranefilters.

11407Z-47----SCM white | black, 0.2 m114H6Z-47----SCM white | black, 0.45 m

High Flow11406Z-47----SCM white | black, 0.45 m139H6Z-47----SCM white | green, 0.45 m

High Flow13906Z-47----SCM white | green, 0.45 m13806Z-47----SCM green | dark green,

0.45 m13006Z-47----SCM gray | white, 0.45 m130H6Z-47----SCM gray | white, 0.45 m

High Flow13005Z-47----SCM gray | white, 0.65 m15407Z-47----SCM green | dark green,

0.2 m

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AirPort MD8AirPort MD8 uses the gelatin membranefilter method guaranteeing reliable andexact measurement results. It is battery-powered and portable for universal use.

16757 AirPort MD8, 100-240 V,47-63 Hz, completewith holder and batterycharger

17528-80-ACD Gelatin membranes,individually sterile-packaged, each in 1 bag

17528-80-BZD Gelatin membranes,individually sterile-packaged, each in 3 bags

arium Laboratory Water SystemsA choice of more than 70 arium versions isavailable to meet all your requirements owater quality and to cover any applicatioWhether for standard applications, routinanalysis or critical applications wherereagent-grade water is required, the arium series consistently supplies highest watequality for your application.

proDI Standard applications, e.g. buffpreparation

proUV Low TOC applications e.g. HPproUF Low endotoxin applicationsproVF Low TOC and low endotoxin

applications

Further laboratory water systems onrequest.

Absorbent PadsThe 1.4 mm thick absorbent pads arewetted with the appropriate liquid culturemedium before a membrane filter is placedon. Each box contains 1,000 absorbentpads in 10 tubes, each with 100 pads,and with manual dispensing device, allpresterilized.

15410-47-ALR Absorbent pads, 47 mm,each approx. 3 mlabsorbent capacity

15410-50-ALR Absorbent pads, 50 mm,each approx. 3.5 mlabsorbent capacity

13906-47-APR Absorbent pads, 47 mm,including membranefilters 0.45 m, white |green grid, individuallysterile-packaged

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Detection Target and Reference 1) Test Sample Materials Media Type (pH)Order No. (Filter Type) 2), 3)

Recom.Incubtion Conditions 4)

Counting of Total Colony Forming Units

Total countAPHA (dairy), APHA (food), APHA (water),AOAC, DAB, EG 98/83, EP, FDA, IDF,ISO 7704, ISO 8199, ISO 9308-1 [1990],ISO 9308-1 [2001], USDA, USP.

Pharmaceuticals, cosmetics, raw materials,water (general quality), waste water, foods,other products.

Caso (pH 7.3)14063--47------N (1)

Bacteria:< 3 d at 3035C;

Yeasts and molds:< 5 d at 3035C

Total countAPHA (water), EP, ISO 7704. Water for pharma purpose, water

(general quality), waste water,other products.

R2A (pH 7.2)14084--47------N (1)14084--47----RDN

> 5 d at 3035C

Total countAPHA (water), ISO 7704, VLB. Raw materials, water (general quality),

waste water, beverages, beer, foods,other products.

Standard (pH 7.2)14064--47------N (1)

< 5 d at 3035C


natural water, waste water, beverages, beer,foods, other products.

Standard TTC (pH 7.2)14055--47------N (1)

< 5 d at 3035C


natural water, waste water, beverages, beer,foods, other products.

Standard TTC I mod. (pH 7.2)14085--47------N (1)14085--47----RDN

< 5 d at 3035C

Total countAPHA (dairy), APHA (food), APHA (water),API, ISO 7704.

Raw materials, water (general quality),natural water, waste water, beverages, softdrinks, concentrates, foods, other products.

TGE| Tryptone GlucoseExtract (pH 7.0)14076--47------N (1)14076--47----RDN

< 5 d at 3035C

Total count

EG 98/83, HMSO, ISO 6222, ISO 7704,ISO 8199. Water (general quality), natural water,other products. Yeast Extract (pH 7.2)14090--47------N (1) 44 4 h at 36 2C;68 4 h at 22 2C

E. Coli and Coliforms, EnterobacteriaE. coli and coliformsISO 7704, Journal Food Protection,ZenHyg (journal of hygiene).

Raw materials, water (general quality),waste water, beverages, foods,other products.

CHROMOCULT* (pH 7.0)14087--47------N (7)14087--47----RDN

2028 h at 36 2C

E. coliAPHA (water), DIN 10110, EG 98/83,ISO 7704, ISO 8199, ISO 9308-1 [2001],LMBG, USDA.


ECD (pH 7.0)14082--47------N (2)

1618 h at 44 2C

E. coli and coliformsAPHA (dairy), APHA (food), APHA (water),

DGHM, ISO 7704, ISO 9308-1 [1990],MNO, USDA.

Raw materials, water (general quality),

natural water, waste water, beverages,soft drinks, concentrates, fruit juice, sugar,sugar products, foods, other products.

Endo (pH 7.4)14053--47------N (9)14053--47----RDN

1824 h at 36 2C

E. coli and coliformsAPHA (food), APHA (water), AOAC, EPA,FDA, ISO 7704, ISO 9308-1 [1990], USDA.


m FC (pH 7.4)14068--47------N (2)14068--50----PDN(closed petri dishes) (2)

1824 h at 36 2C

Enterobacteria, E. coliAPHA (dairy), APHA (food), APHA (water),AOAC, DAB, DIN 38411, DGHM,EP, ISO 7704, LMBG, MNO, USDA, USP.

Pharmaceuticals, cosmetics, raw materials,water (general quality), natural water, wastewater, beverages, soft drinks, concentrates,fruit juice, foods, other products.

MacConkey (pH 7.1)14097--47------N (2)

1872 h at 3035C

Technical Data and Application Guide Nutrient Pad Sets

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Shelf Life Test Strains 5)

24 01, 03, 05, 09, 18, 22, 25, 26

24 01, 03, 05, 09, 18, 22, 26

24 03, 07, 09, 18, 26

24 03, 07, 09, 18, 26

24 03, 07, 09, 18, 26

24 09, 18, 26

24 03, 07, 09, 18, 26

24 06, 09, 11, 21, 25

24 06, 09, 19, 21, 22

24 06, 07, 09, 21, 25, 28

24 06, 07, 09, 11, 21

24 02, 06, 09, 21, 25, 26

1) Reference Guide on page 34.

2) A Set contains 100 Nutrient Pads and100 membrane lters, both individually, sterilepackaged. The membrane lters are selectedfor optimum growth together with the

corresponding nutrient media. The suppliedmembrane lter type is listed within brackets:

(1) = green with dark green grid,0.45 m pore size

(2) = white with green grid, 0.45 m pore size (3) = gray (after wetting black) with white grid,

0.65 m pore size (4) = white with green grid, 0.65 m pore size (5) = white with green grid, 1.2 m pore size (6) = gray (after wetting black) with white grid,

0.8 m pore size (7) = white with black grid, 0.45 m pore size (8) = gray (after wetting black) with white grid,

0.45 m pore size (9) = white with green grid, 0.45 m pore size,

High Flow (ideal for E. coli)

(10) = gray (after wetting black) with white grid,0.45 m pore size, High Flow

3) Diameter of the membrane lter, 47 mm.Order number for Nutrient Pad Setswith 50 mm membrane lter as above, but--47------N replaced by --50------N.

Most of the NPS types are also available withMicrosart e.motion Membrane Filters:Order number as above, but ---N replaced by-RDN.

Other NPS types and NPS withMicrosart e.motion Membrane Filters on request.

4) The incubation conditions are recommendedby Sartorius Stedim Biotech. They may be variedaccording to the type of samples in compliancewith the reference standard or customersrequirements.

5) Test strains on page 32.

* Trade mark owner and manufactureris Merck KGaA

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E. coli and coliformsAFNOR, APHA (water), BS, FDA, ISO 7704,ISO 9308-1 [1990], USDA.

Water (general quality), waste water,beverages, foods, other products.

Teepol|Lauryl Sulphate (pH 7.2)

14067--47------N (2)14067--47----RDN

1824 h at 36 2C

E. coli and coliformsAPHA (food), EG 98/83, ISO 7704,ISO 8199, ISO 9308-1 [1990],ISO 9308-1 [2001].


Tergitol TTC (pH 8.0)14056--47------N (2)14056--47----RDN

1824 h at 362C

Other Faecal BacteriaEnterococciAPHA (food), APHA (water), EG 98/83,HMSO, ISO 7704, ISO 7899-2, ISO 8199,LMBG, MNO.

Raw materials, water (general quality),natural water, waste water, beverages,foods, other products.

Azide| KF Strep (pH 7.2 0.1)14051--47------N (1)14051--47----RDN

4048 h at 36 2C

SalmonellaeAFNOR, APHA (dairy), APHA (food), AOAC,DGHM, FDA, HMSO, IDF, ISO 6579 [1981],ISO 7704, USDA.


Bismuth Sulfite (pH 7.6)14057--47------N (1)

4048 h at 36 2C

Non-faecal, Pathogenic BacteriaPseudomonas aeruginosaAPHA (water), AOAC, ASM, DIN 38411,EG 98/83, FDA, ISO 7704, ISO 8199,ISO 16266.

Cosmetics, raw materials, water(general quality), waste water, foods,other products.

Cetrimide (pH 7.1)14075--47------N (2)14075--47----RDN

4048 h at 362C

Staphylococci, Staph. aureusAPHA (food), AOAC, DGHM, FDA, HMSO,ISO 7704, USP.


Chapman (pH 7.4)14074--47------N (2)

1872 h at 3035C

Yeasts and MoldsWild yeastsJournal Institute of Brewing, VLB. Beer, other products. Lysine (pH 5.0)

14061--47------N (3)35 d at 3035C

Yeasts and moldsAPHA (food), AOAC, IFU. Beverages, wine, soft drinks, concentrates,

fruit juice, foods, other products.Malt Extract (pH 4.5)14086--47------N (6)14086--47----CCN (8)

35 d at 2025C or at3035C depending on thetarget of the investigation

Yeasts and moldsAPHA (food), AOAC, EP, USP. Pharmaceuticals, cosmetics, raw materials,

water (general quality), waste water,other products.

Sabouraud (pH 5.6)14069--47------N (10)

< 5 d at 2025C

Yeasts and molds

Wine, soft drinks, concentrates, sugar,sugar products, other products. Schaufus Pottinger |m Green yeast and mold (pH 4.3)14070--47------N (4)14072--47------N (5)14080--47------N (6)14080--47----RDN14083--47------N (3)14091--47------N (8)14091--47----RDN

25 d at 2025C or at3035C depending on thetarget of the investigation

Yeasts and molds and bacteriaISO 7704. Beverages, beer, wine, soft drinks,

concentrates, fruit juice, other products.Wallerstein | WL Nutrient (pH 5.5)14089--47------N (2)

25 d at 3035C aerobic oranaerobic depending on thetarget of the investigation

Yeasts and molds VLB. Raw materials, beverages, beer,wine, soft drinks, concentrates, foods,other products.

Wort (pH 4.4)14058--47------N (3)14092--47----RDN (8)

35 d at 2025C or at3035C depending on thetarget of the investigation30


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24 06, 07, 09, 11, 21

24 06, 07, 09, 11, 21

24 07, 08, 09, 22, 26

24 03, 09, 21, 25, 26

24 04, 09, 21, 22, 26, 30

24 07, 09, 21, 26, 27

24 05, 20, 23, 24

24 05, 20, 23, 24

24 01, 05, 20, 23, 24

24 03, 05, 20, 23, 24

24 05, 12, 19, 20, 23

24 05, 20, 23, 24
















31


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Product-spoiling MicroorganismsThermophilic spore formers andmesophilic bacteriaAPHA (dairy), APHA (food), AOAC,ICUMSA, IFU, ISO 7704, NCA.

Fruit juice, sugar, sugar products, foods,other products.

Glucose Tryptone (pH 6.8)14066--47------N (2)

1872 h at 3035C formesophilic bacteria;4872 h at 552C forthermophilic sporulatingmicroorganisms

Leuconostoc oenos and other winespoiling organ.ISO 7704, Lanaridris& Lafon-Lafourcade.

Wine, fruit juice, other products. Jus de Tomate |Tomato Juice (pH 5.0)14079--47------N (1)

57 d at 3035C anaerobic(microaerophil); controlfor slowly growing micro-organisms after 10 d isrecommended

Acid-tolerant microorganismsAPHA (water), IFU, ISO 7704,MPP (packaging staff).

Raw materials, water (general quality),waste water, wine, soft drinks, concentrates,

fruit juice, foods, other products.

Orange Serum (pH 5.5)14062--47------N (1)

14062--47----RDN

35 d at 3035C aerobic oranaerobic depending on the

target of the investigationAcid-tolerant microorganismsAPHA (water), IFU, ISO 7704,MPP (packaging staff).

Raw materials, water (general quality),waste water, wine, soft drinks, concentrates,fruit juice, foods, other products.

Orange Serum (pH 3.2)14096--47------N (6)14096--47----RDN

35 d at 3035C aerobic oranaerobic depending on thetarget of the investigation

Lactobacilli and Pediococci andother beer spoiling organismsEBC, ISO 7704, MEBAC, VLB.

Beer, other products. VLB-S7-S (pH 5.5)14059--47------N (2)

35 d at 3035C anaerobic(microaerophil)

LactobacilliAPHA, ISO Fruit juice, beer, diary, foods, soft drinks,

other materials.MRS (pH 6.1)1407747------N (1)

35 d at 30C under anaerobconditions (microaerophil)

Test Strains [ATCC No.], [DSM No.]

01. Aspergillus brasiliensis 16404, 198802. Bacillus cereus 11778, 34503. Bacillus subtilis subsp. spizizenii 6633, 34704. Brevundimonas diminuta 19146, 163505. Candida albicans 10231, 138606. Enterobacter aerogenes 13048, 30053

07. Enterococcus faecalis 29212, 257008. Enterococcus faecium 19434, 2047709. Escherichia coli 8739, 157610. Geobacillus stearothermophilus

7953, 593411. Klebsiella pneumoniae 13883, 3010412. Lactobacillus lindneri DSM 2069013. Lactobacillus plantarum subsp. plantarum

14917, 2017414. Leuconostoc mesenteroides subsp.

mesenteroides 8293, 2034315. Oenococcus oeni 23279, 20252

17. Raw cane sugar solution (10%)18. Tap water19. Pediococcus damnosus 29358, 2033120. Penicillium commune 10428, 221121. Proteus mirabilis 29906, 447922. Pseudomonas aeruginosa 9027, 1128

23. Rhodotorula mucilaginosa DSM 7040324. Saccharomyces cerevisiae 9763, 133325. Salmonella enterica subsp. enterica

serotype typhimurium 14028, 1958726. Staphylococcus aureus subsp. aureus

6538, 79927. Staphylococcus epidermidis 12228, 179828. Escherichia coli 25922, 110329. Lactobacillus brevis 14869, 2005430. Pseudomonas aeruginosa 27853, 1117

Mesophilic slime-forming bacteriaesp. Leu. MesenteroidesICUMSA, ISO 7704.

Soft drinks, concentrates, sugar,sugar products, other products.

Weman (pH 5.5)14065--47------N (1)

35 d at 3035C

31. Lactobacillus acetotolerans 43578, 20749

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24 02, 09, 10, 17, 26

24 12, 14, 15, 24

24 02, 05, 13, 14, 20, 23, 24

24 02, 05, 13, 14, 20, 23, 24

24 06, 12, 13, 19, 24, 29

24 13, 19, 22, 29, 31
















24 14

RemarksThe incubation conditions are recommended by Sartorius Stedim Biotech. They may be variedaccording to the type of samples in compliance with the reference standard or customersrequirements.The description of the typical results or any pictures show typical appearance of the mentionedmicroorganisms. In particular cases, color and shape of the colonies could vary from the expectedhabitus. Further tests may be necessary to validate the result.Sartorius Stedim Biotech shall not be liable for consequential and|or incidental damage sustainedby any customer from the use of its products.Nutrient Pad Sets (NPS) are subject to continuous product improvement as part of ourproduct development program to align our products with changing application requirements.For current specifications and lot release criteria please visit our homepage under:www.sartorius-stedim.com/NPSSearch.

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Reference GuideThe compositions of the pads are based on the recommendations of numerous different standards and regulations. Abbreviation TitleAFNOR Association Franchaise de Normalisation

APHA (dairy) American Public Health Association: Standard Methods for the examination of dairy productsAPHA (food) American Public Health Association: Compendium of methods for the microbiological examination of foodAPHA (water) American Public Health Association, American Water Works Association (AWWA) and Water Environmen

Federation (WEF): Standard Methods for the Examination of Water and Waste WaterAOAC Association of Official Analytical ChemistsAPI American Petroleum Institute: Recommended practice for biological Analysis of Subsurface Injection watersASM American Society for MicrobiologyBS British StandardsDAB Deutsches Arzneimittelbuch (German Pharmacopoeia, replaced by EP)DIN 10110 Deutsches Institut fr Normung: Mikrobiologische Fleischuntersuchung. Bestimmung der E. coli.

(Microbial detection of E. coli on meat)DIN 38411 Deutsches Institut fr Normung: Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlamm-

untersuchung (German standard for water, waste water and sludge analysis)DGHM Deutsche Gesellschaft fr Hygiene und Mikrobiologie (German Association of Hygiene and Microbiology)EBC European Brewery ConventionEG 98/83 European Guideline 98/83: Water Quality for human purposeEP European PharmacopoeiaEPA U.S. Environmental Protection Agency: Laboratory standards for equipment and materialsFDA U.S. Federal Drug AdministrationHMSO Her Majestys Stationery Office: Department of Health and Social Security (1982) The Bacteriological

Examination of Drinking Water Supplies. Report 71, HMSO, LondonICUMSA International Commission for Uniform Methods of Sugar AnalysisIDF International Dairy FederationIFU International Federation of Fruit Juice ProducersISO 6222 International Organization for Standardization: Water Quality - Enumeration of culturable micro-organismsISO 6579-1981 International Organization for Standardization: Microbiology. General guidance on methods for the

detection of Salmonella. Reference methodISO 7704 International Organization for Standardization:

Water Quality, Evaluation of membrane filters used for microbiological analysisISO 7899-2 International Organization for Standardization:

Water Quality Detection and enumeration of intestinal enterococciISO 8199 International Organization for Standardization:Water Quality General Guide to the enumeration of micro-organisms by culture

ISO 9308-1 International Organization for Standardization:Water Quality Detection and enumeration of E. coli and coliform bacteria

EN ISO 16266 European |International Organization for Standardization:Water Quality Detection and enumeration of Ps. aeruginosa

JFoodP Journal of Food ProtectionJIBrew The Journal of the Institute of BrewingLLL Method described by Lanaridris& Lafon-LafourcadeLMBG Amtliche Sammlung von Untersuchungsverfahren nach dem 35 des Lebensmittel- und Bedarfs-

gegenstndegesetzes des BGA (testing procedures for food stuffs and articles of daily use)

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Abbreviation TitleMEBAK Methodensammlung der Mitteleuropischen Brauereitechnischen Analysenkommission

(methods of the Central European brewery commission)MNO Verordnung ber natrliches Mineralwasser, Quellwasser und Tafelwasser (Mineral/Table Water Guidel

MPP Merkbltter fr die Prfung von Packmitteln (Testing procedures for packaging stuff)NCA National Canners Association: A Laboratory manual of the canning industryUSDA U.S. Department of AgricultureUSP United States Pharmacopoeia VLB Versuchs- und Lehranstalt fr Brauerei in Berlin (institute of brewery)ZenHyg Zentralblatt fr Hygiene (Journal of Hygiene)

DIN standards and the Amtliche Sammlung von Untersuchungsverfahren nach dem35 des Lebensmittel- und Bedarfsgegenstndegesetzes des BGA are available through the Germanpublisher Beuth-Verlag, Burggrafenstr. 6, 10787 Berlin

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Sales and Service ContactsFor further contacts, visit www.sartorius.com

EuropeGermanySartorius Stedim Biotech GmbHAugust-Spindler-Strasse 1137079 GoettingenPhone +49.551.308.0Fax +49.551.308.3289

Sartorius Stedim Systems GmbHRobert-Bosch-Strasse 5 734302 GuxhagenPhone +49.5665.407.0Fax +49.5665.407.2200

FranceSartorius Stedim FMT S.A.S.ZI Les PaludsAvenue de Jouques CS 9105113781 Aubagne CedexPhone +33.442.845600Fax +33.442.845619

Sartorius Stedim France SASZI Les PaludsAvenue de Jouques CS 7105813781 Aubagne CedexPhone +33.442.845600Fax +33.442.846545

AustriaSartorius Stedim Austria GmbHFranzosengraben 121030 ViennaPhone +43.1.7965763.18Fax +43.1.796576344

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1800 VilvoordePhone +32.2.756.06.80Fax +32.2.756.06.81

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NetherlandsSartorius Netherlands B.V.Phone +31.30.60.53.001Fax [email protected]

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SwitzerlandSartorius Stedim Switzerland AGRingstrasse 24 a8317 TagelswangenPhone +41.52.354.36.36Fax +41.52.354.36.46

U.K.Sartorius Stedim UK Ltd.Longmead Business CentreBlenheim Road, EpsomSurrey KT19 9 QQPhone +44.1372.737159Fax +44.1372.726171

AmericaUSASartorius Stedim North America Inc.5 Orville Drive, Suite 200Bohemia, NY 11716Toll-Free +1.800.368.7178Fax +1.631.254.4253

ArgentinaSartorius Argentina S.A.Int. A. valos 4251B1605ECS MunroBuenos AiresPhone +54.11.4721.0505Fax +54.11.4762.2333

BrazilSartorius do Brasil LtdaAvenida Senador Vergueiro 2962So Bernardo do CampoCEP 09600-000 - SP- BrasilPhone +55.11.4362.8900Fax + 55.11.4362.8901

MexicoSartorius de Mxico S.A. de C.V.Circuito Circunvalacin PonienteNo. 149Ciudad Satlite53100, Estado de MxicoMxicoPhone +52.5555.62.1102Fax +52.5555.62.2942

Asia|PacificAustraliaSartorius Stedim Australia Pty. Ltd.

Unit 5, 7-11 Rodeo DriveDandenong South Vic 3175Phone +61.3.8762.1800Fax +61.3.8762.1828

ChinaSartorius Stedim Biotech (Beijing) Co. Ltd.No. 33 Yuan RoadAirport Industrial Park Zone BShunyi District, Beijing 101300Phone +86.10.80426516Fax +86.10.80426580

Sartorius Stedim Biotech (Beijing) Co. Ltd.Shanghai Branch Office3rd Floor, North Wing, Tower 1No. 4560 Jin Ke RoadPudong District, Shanghai 201210Phone +86.21.68782300Fax +86.21.68782332 | 68782882

Sartorius Stedim Biotech (Beijing) Co. Ltd.Guangzhou Representative OfficeUnit K, Building 23Huihua Commerce & Trade BuildingNo. 80 Xianlie Middle RoadGuangzhou 510070Phone +86.20.37618687 | 37618651Fax +86.20.37619051

IndiaSartorius Stedim India Pvt. Ltd.#69/2-69/3, NH 48, JakkasandraNelamangala Tq562 123 Bangalore, IndiaPhone +91.80.4350.5250Fax +91.80.4350.5253

JapanSartorius Stedim Japan K.K.4th Fl., Daiwa Shinagawa North Bldg.8-11, Kita-Shinagawa 1-chomeShinagawa-ku, Tokyo, 140-0001 JapanPhone +81.3.4331.4300Fax +81.3.4331.4301

MalaysiaSartorius Stedim Malaysia Sdn. Bhd.Lot L3-E-3B, Enterprise 4Technology Park MalaysiaBukit Jalil57000 Kuala Lumpur, Malaysia

Phone +60.3.8996.0622Fax +60.3.8996.0755

SingaporeSartorius Stedim Singapore Pte. Ltd.1 Science Park Road,The Capricorn, #05-08A,Singapore Science Park IISingapore 117528Phone +65.6872.3966Fax +65.6778.2494

South KoreaSartorius Korea Biotech Co., Ltd.8th Floor, Solid Space B/D,PanGyoYeok-Ro 220, BunDang-GuSeongNam-Si, GyeongGi-Do, 463-400

Phone +82.31.622.5700Fax +82.31.622.5799

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