MICROBIOLOGY SAMPLING GUIDE - EMSL Analytical, Inc. · LABORATOR Y TESTING SERV ICES SINCE 1981...

L A B O R A T O R Y T E S T I N G S E R V I C E S S I N C E 1 9 8 1

M I C R O B I O L O G Y S A M P L I N G G U I D E

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LAB Services Include:Asbestos, Mold, Bacteria, Industrial Hygiene, Metals, Environmental Chemistry,

Food & Conumer Products, Allergens, PCR-Polymerase Chain Reaction (DNA),

Silica, Volatiles Scan, Formaldehyde by HPLC, Indoor Air Quality, Water,

Radon and Materials Testing.

www.emsl.comemail: [email protected]

UNITED STATES

C A N A D A

Midwest866.736.4824877.435.5227

North East 800.220.3675

South East888.958.8170877.476.5866

South Central866.318.3920

Toronto289.997.4602

West Coast888.455.3675866.798.1089

Headquarters

EMSL Laboratory Location

EMSL Service Center Location

LA Testing Laboratory Location

EMSL Analytical Locations

For more information contact EMSL Analytical, Inc. 1-800-220-3675 • www.emsl.com • Contact EMSL for all your Sampling Supplies and Cassettes

C O N T E N T S Page

Air Sampling for Total Spore Counts Air-O-Cell 4 MoldSnap & MICRO-5 5 Allergenco-D 5

Culturable Air Sampling (Fungi or Bacteria) 6-7

Surface Sampling 8-10 (Tape Lifts, Bulk Samples, Swabs)

ERMI Dust Sampling 11

Real Time PCR Sampling 12

Allergen Sampling 13 Water Sampling for Total Coliforms, E. Coli 14 & Fecal Coliform

Sewage Contamination in Buildings 15

Legionella Sampling 16-17

Methicillin-resistant Staphylococcus aureus 18 (MRSA) Sampling

Endotoxin Sampling 19

Food Sampling 20-21

Limiting Conditions for Pathogen Growth 23

Microbiology Culture Media Information 24-25

EMSL Lab Locations 26

3

For more information contact EMSL Analytical, Inc. 1-800-220-3675 • www.emsl.com • Contact EMSL for all your Sampling Supplies and Cassettes4

Air Sampling for Total Spore Counts

AIR-O-CELL • TheAir-O-Cell™cassette(ProductID8715301B)isasingle-usesamplingdevice designed for the rapid collection and analysis of a wide range of airborne particles. These include fungal spores, pollen, insect parts, skin cell fragments, fibers, and other inorganic particulates. • Thecassetteisdesignedtooperateataflowrateof15LPM.Lowerflowrates may result in a collection loss of some spores and the accumulation of others in a non-uniform manner. Therefore, it is critical to run the sampling pumps at the manufacturer’srecommendedairflowrate.

Benefits • Usefulforinitialsitetesting,especiallyiffungalgrowthisnotvisible. • Quickandsimpleprocedure. • Fastturnaroundtimesavailable. • Lowchanceofsamplecontamination. • Necessaryfordeterminingallergicmoldsporepotential. Moldsporescancauseallergieswhethertheyareviable ornon-viable

Disadvantages • Fungicannotbeidentifiedtospecieswiththismethod. • Duetothesimilaritiesinsporemorphology,somesporeswillbe grouped together, i.e., Aspergillus sp. and Penicillium sp. • Sporeviabilitycannotbeassessed.

Sampling Procedure 1. Priortosampling,calibrateyourpumpto15litersperminuteviaarotameter.Itisalsorecommendedthat therotameterorsamplingpumpbeperiodicallycalibratedtoaNISTprimarystandard.IfusingtheZefon Bio-Pump(ProductID8706002)usethespecificallydesignedAir-O-Cellflowindicator,aspeciallydesigned bubbletubeNISTPrimarycalibrator(ProductID8704200).Itcannotbeusedwithanyothertypeofpump. 2. Removeandretaintapesealcoveringinletandoutletonthecassette. 3. Attachtheoutlet(roundhole)toastandard1/2”PVCtubing(forusewithhighvolumepumpsonly). 4. Start the sampling pump and sample for an appropriate period of time (see recommendations below). 5. RemoveAir-O-Cellfromtubingandresealwiththeoriginalseals.Labelsample. 6. CompleteanEMSLChainofCustody(COC),availableatwww.emsl.com,detailingclientnameand information,projectnameornumber,sample#,descriptionofarea,andvolumeofaircollected. 7. Toreduceshippingdamage,itisrecommendedthattheAir-O-Cellbeplacedinacorrugatedboxwith paddingtoensuresafearrivalatthelaboratory.

Sampling Duration • Thesamplingtimeisdependentonthedensityofparticulateintheenvironment.Itisimportantnotto overloadthesample,otherwiseitwillbeimpossibletoaccuratelycountthetypesofspores,pollenorother particulates that are present. The following list represents typical sample times to attain a sharply defined trace with good dispersion of the spores: 1.Clean“office”oroutdoors(novisibledust)=10minutes 2.Indoorenvironment,highactivity&personnel=5minutes 3.Indoorenvironment,drywallrenovationorheavyindustrialdust=1minute.

Quality Control Recommendations • Aneffectiveinterpretationisbasedonthecomparisonofindoorandoutdoorsamples.Outdoorsampleswill help demonstrate whether spore amplification is occurring indoors. • Obtainacontrolsamplefromanon-complaintareaforcomparison. • Sendingablankcassetteforanalysisperprojectisagoodpractice. • Flowrateiscriticalforaccurateresults.Remembertocalibrateandrecalibratethepumppriortoallsampling. (15 liters/minute) • Neverusecassettesthataredamagedorexpired.

FormoreinformationcontactZefonInternationalat:www.zefon.com. Air-O-Cell™ is a registered trademark of Zefon International.

Air-O-Cell 50 Pack, #8715301B

List Price: $199.00 USD

10 Pack, #8715302 List Price: $55.00USD

Bio-Pump Plus #8706002 List Price: $729.00 USD

For more information contact EMSL Analytical, Inc. 1-800-220-3675 • www.emsl.com • Contact EMSL for all your Sampling Supplies and Cassettes

MoldSnap & MICRO-5• TheZefonMoldSnap(ProductID8715304)andtheMicro5MicroCell (ProductID8715306)weredesignedtooperateataflowrateof5litersper minute for optimal collection efficiency. Cassettes should be stored at 50° - 80° F. • Donotuseintemperaturesbelow37°F!

Sampling Procedures 1. RemovepinfromthebottomoftheMoldSnaporMicro5. 2. Ifusingaconventionalhighvolumepump:Simplyconnectoneendoftubing (5 feet or less) to the bottom of the cassette. 3. IfusingalowvolumepumpsuchasaGilAir5:Simplyattacha1”pieceoftubing to the bottom of the cassette. 4. Connecttheotherendoftubingtoapumppre-calibratedtoaflowrateof5LPM. 5. Removecapfromthetopofthecassette. 6. Turn pump on and take a sample for an appropriate amount of time depending on environment.(See“SamplingDuration”below.) 7. When sampling is completed, replace the pin & cap to the bottom and top of the cassette.

Sampling Duration • Outdoor/IndoorCleanEnvironments=8-10minutes • Indoor(NormalActivity)=5minutes • Indoor(HeavyParticulates)=1-3minutes • InnerwallcavityusingtheMicro5only (withoptionalInnerWallAdapter,ProductID8715901)=1-2minutes

Allergenco-D• TheAllergenco-D(ProductID8715307)wasdesignedtooperateataflowrate of 15 liters per minute for optimal collection efficiency.

Sampling Procedure 1. RemovesealfromthebottomoftheAllergenco-D. 2. Simply connect one end of tubing (5 feet or less) to the bottom of the Allergenco-D. 3. Connecttheotherendoftubingtoapumppre-calibratedtoaflowrateof15LPM. 4. RemovesealfromthetopoftheAllergenco-D. 5. Turnpumponandcollectasampleforupto10minutes,dependingonenvironment (See “Sampling Duration”). 6. When sampling is complete, replace seals to the bottom and top of the cassette.

Sampling Duration • Outdoorsample1-10minutes • Dust-freeenvironment(cleanoffice)5-8minutes • Indoorenvironment(occupiedspace)3-5minutes • Indoorenvironment(excessvisibledust)1-3minutes • Innerwallsample1-5minutes(usingoptionalwalladapter,ProductID8715908)

5

Tech Tip: Basic Mold Cleanup

The key to mold control is moisture control. It is important to dry water damaged areas and items within 24-48hourstopreventmoldgrowth.Ifmoldisaprobleminyourhome,cleanupthemoldandgetridofthe excess water or moisture. Fix leaky plumbing or other sources of water. Wash mold off hard surfaces with detergent and water, and dry completely. Absorbent materials (such as ceiling tiles & carpet) that becomemoldymayhavetobereplaced.(Source:EPAhttp://www.epa.gov/mold/moldresources.html)

Mold Snap 50 Pack, #8715304B


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Bios Defender 510-H HighFlow

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6 For more information contact EMSL Analytical, Inc. 1-800-220-3675 • www.emsl.com • Contact EMSL for all your Sampling Supplies and Cassettes

CulturableAirSampling(FungiorBacteria)

ParticleImpactors(Andersen-typeSamplers) • ThismethodofairsamplinginvolvesdrawingameasuredvolumeofairoverculturemediainPetridishes. The Petri dishes are incubated in the laboratory so the organisms impacted on the plate can grow. The fungi orbacteriaarecountedandidentified.ThismethodcommonlyusesanAndersenN-6typeimpactor(e.g.EMSL VP-400MicrobialSamplerProductID8709001).DifferentagarplatesareavailablefromEMSLAnalytical, Inc., depending on the types of fungi or bacteria to be identified.

Benefits • Fungalculturescandeterminewhetherthefungusisviable(alive),andallowsforgenusandspecies identification. • Bacterialculturesprovideenumerationandidentificationofculturablebacteriapresentintheair.

Disadvantages • Culturestake6-10daysforthemicroorganismstogrowandbeidentified. • Sincemostenvironmentalsamplescontainalargenumberoforganisms,eachhastocompetewithothersto grow on the media. As a result, fungi and bacteria present in the air may not be as well represented in culture. • Somemicrobesdonotgrowwelloratallontheculturemedia. • Somemicroorganismsareunabletobeidentified,astheyfailtoproducekeycharacteristicssuchassporesor they may not be described in the scientific literature.

GeneralMediaRecommendationsFungiandBacteria • Forfungalsampling,ingeneral,werecommendMaltExtractAgar(MEA). • Ifyouaresamplingindryareas,theuseofDG18willhelpselectforthegrowthofdry-lovingfungithat may not grow on MEA agar • SamplingforStachybotryssp.canbeachievedwitheitherCelluloseAgar(CA)orCornmealAgar(CMA). • Forbacterialsampling,ingeneral,werecommendTrypticSoyAgar(TSA)orTSAw5%blood. • ForsamplingGramnegativebacteria,werecommendMacConkeyAgar(MAC) • Forallothersituations,theMicrobiologyDepartmentwillbehappytomakerecommendationsbasedon yourindividualsamplingsituation. • Samplingsuppliesmaybeorderedatwww.emsl.comorbycallingCustomerService:800-220-3675.

HowtoHandleMicrobiologicalMedia(Agarplates) • Agarplatesmustbekeptrefrigeratedoronfreezerpacksuntilreadytouse. • Theplatesmustbeallowedtowarmuptoroomtemperaturebeforetakingasample(approx.15minutes). • Donotremovethelidfromtheplateatanytimeexceptduringsampling. • SealthelidtotheplateaftersamplecollectionwithParafilmortape. • TheplatesmustbeshippedbacktoEMSLwithfreezerpacksbyOVERNIGHTPRIORITY. Refreezeandreusetheoriginalfreezerpack(thistypeoffreezerpackisstablefor24hours). • Adequatepackingmaterialmustbesenttoprotecttheplates. • Platesmustnotcomeintodirectcontactwiththefreezerpack,asthemediamayfreeze,invalidatingthetests. • IfthereisanydelayinsendingtheagarplatestoEMSL,theyshouldberefrigerateduntilreadyfor overnightdelivery.

Recommendations • Wearlatexornitrileglovesduringsampling. • Use70%isopropylalcoholtodisinfectsamplingdevicebetweeneachsample. • PlacePetridishlidinacleanbagduringsamplingtoreduceanycrosscontamination. • Includeoutsidesamplesandafieldblankforcontrol.

For more information contact EMSL Analytical, Inc. 1-800-220-3675 • www.emsl.com • Contact EMSL for all your Sampling Supplies and Cassettes 7

Sampling Procedure 1. Allow agar plates to reach room temperature before use. 2. Attachoneendoftubingtotheintakeofthevacuumpumpandtheotherendtotheinletofthesampler. 3. Calibratetheflowrateofthevacuumpump: a.PlaceanuncoveredPetridishintosampler(Donotsubmitthisdishasasample,discardaftercalibration). b.Turnonpumpandadjustflowuntiltherotameterisat28.3LPM (flowrateisreadfromthemiddleofbearingontherotameter). c.Removestopperandrotameterpriortosampling. 4. Wipeallexposedsurfacesofsamplerwitha70%isopropylalcoholpadandallowtoairdry. 5. Place the agar plate on the sampler base so that the Petri dish rests on the three raised metal pins. 6. RemovethecoverofthePetridishandplaceintoacleansamplebagtominimizecontamination (availableuponrequest). 7. Assemble the jet classification stage on the sampler and secure the inlet cone with the three attached clips. 8. Settimertoappropriatetimedependingonenvironmentalconditions(samplingtimeisusually between 2-5 minutes). 9. Turn on the pump and start the timer simultaneously. 10. When the time is up, turn off the pump and disassemble sampler and placecoverbackontoagarplate. 11. Secure lid onto Petri dish with masking tape or Parafilm (avoidusingelectrical,packing,transparentandducttape). 12. Write the sample number on the bottom of the Petri dish. 13. Record all appropriate information on the Chain of Custody. 14. Return samples with an ice pack to EMSL Analytical for analysis.

Tech Tip: Canmoldcausehealthproblems?

Molds are usually not a problem indoors, unless mold spores land on a wet or damp spot and begin grow-ing.Moldshavethepotentialtocausehealthproblems.Moldsproduceallergens(substancesthatcancauseallergic reactions), irritants, and in some cases, potentially toxic substances (mycotoxins). Inhaling or touch-ingmoldormoldsporesmaycauseallergicreactionsinsensitiveindividuals.Allergicresponsesincludehayfever-typesymptoms,suchassneezing,runnynose,redeyes,andskinrash(dermatitis).Allergicreactionsto mold are common. They can be immediate or delayed. Molds can also cause asthma attacks in people with asthma who are allergic to mold. In addition, mold exposure can irritate the eyes, skin, nose, throat, and lungs of both mold-allergic and non-allergic people. Symptoms other than the allergic and irritant types are not commonly reported as a result of inhaling mold. Research on mold and health effects is ongoing.Thisbrochureprovidesabriefoverview;itdoesnotdescribeallpotentialhealtheffectsrelatedtomoldexposure.Formoredetailedinformationconsultahealthprofessional.Youmayalsowishtoconsultyour state or local health department. (Source EPA Document # 402-K-02-003)

EMSL VP-400 BasicKitwith E-Lite Pump 25 Pack, #8709003 List Price: $459.00 USD

EMSL VP-400 Single Stage MicrobialSampler #8709001 List Price: $375.00 USD


Surface Sampling

DirectExamination(TapeLift,Bulk,Swab) • Adirectexamallowsfortherapiddeterminationofthepresenceoffungal spores as well as identifies the types of fungi. • Directexaminationsshouldonlybeusedtosamplevisiblemoldgrowthin acontaminatedareasincemostsurfaceswillhaveadepositoffungalspores thatarenormallypresentintheenvironment.

Benefits • Thedirectexamisinexpensiveandcanbeperformedquickly. • Ausefultestfordeterminingifthereismoldamplification. • Directsamplingmayrevealindoorreservoirsofsporesthat havenotyetbecomeairborne.

Disadvantages • Areasoffungalgrowthareoftensmallandscattered,sotheymaynotallbepickedup.Choosingmultiple samplinglocationswillhelpovercomethisproblem. • Healthproblemsrelatedtoindoormicrobialgrowtharegenerallycausedbytheinhalationofsubstantial numbersofairbornespores,sometimesoveralongperiodoftime.Thepresenceofbiologicalmaterialson a particular surface may not be a direct indication of what is in the air. • Thismethoddetectsbothviableandnon-viablesporesbutcannotdistinguishbetweenthem.Itisadvisable tocombinedirectexamsampleswithculturemethodsifknowingviabilityisimportanttoyourproject. • Tapeliftsarenotabletobecultured. • Ifadirectexaminationofaswabsampleistaken,afollowupbycultureispossible. • Directexaminationsofdirt/soilanddustsamplescannotbeperformedreliablybecauseofpreparationlimitations. • Fungiusuallycannotbeidentifiedtospeciesandsometimesnoteventogenuswiththismethod.Forexample, Aspergillus sp. and Penicillium sp. are normally reported together due to the similarities in spore morphology, unless fruiting structures are present that allows for a better identification.

Materials For Tape Lift: • WerecommendusingEMSLTapekitsorBio-Tapes(ZefonIntl)otherwiseclear (transparent) Scotch or other brand tape (frosted tape obscures the spores). • Newplasticbagtoholdsample(s)(providedinatapeliftkitavailablefromEMSL). • Onlyusetapeliftsonhard,flatsurfaces For Bulk: • Sterilecontainer,newplasticbagorWhirl-Pak(providedbyEMSL) to hold and transport samples. For Swab: • SterileTransPorterswabtocollectandtransportsamples(providedbyEMSL). For all Matrices: • Latex/nitrilegloves(alsocanbeprovidedatyourrequest).

Sampling Procedure

Tape-lift 1.Takeafewinchesofcleartape.Avoidtouchingthestickyside,especiallythe part to be used to collect the mold. 2.Wearinggloves,applythecentralinchoftapetothesuspectarea(chooseone thatisfreeofextraneousdebris).Applylightpressuretothenon-adhesiveside. 3. Pull tape off surface with slow, steady pressure, holding the tape edges only. 4.Applystickysideoftapetotheinsideoftheplasticbag(ziplock). 5. Ensure there are no folds or creases in the tape. 6. Close bag and label appropriately. (Put only one sample in each bag.)

Bio-Tape 25 Pack, #8708325 List Price: $25.00

USD

Nitrile Gloves

Small #8705400 List Price:

$8.95 USD

Medium #8705401 List Price:

$8.95 USD

Large #8705402 List Price:

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X-Large #8705403 List Price:

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Bulk 1. Wearinglatexglovesremoveasmallpieceofthesuspectmaterial(1x1inchpieceismorethansufficient). 2. Placepieceinsidecleansterilecontainerornewplasticbag(ziplock). 3. Close bag or cap container and label appropriately.

Swab 1. Wearinggloves,removeswabfrompackagingmaterial. 2. Removeplugfrommediatube. 3. Swabthedesiredareathoroughly,rollingtheswablightlybackandforthoversamplingarea. 4. Insert the swab in the tube, firmly close cap, and label appropriately. 5. CompleteanEMSLChainofCustody(COC),availableonourwebsite(www.emsl.com),detailing client name and information, project name or number, sample #, and a description of the area.

Quality Control Recommendations For Tape Lift: • Usecleartape--notfrosted,electrical,duct,orpackingtape. • Donotfoldtapeontoitself. • Sticktapeontheinsideoftheplasticbagonly. • Pleasedonotsendtapeonslidesorcoverslips.Theymayarrivebrokenmakingthesampledifficulttoanalyze. For Bulk: • Sendarepresentativeportionofthesample,iflarge.Thispreventsover-handlingofthesampleandcontamination. Ifanalysisofaspecificportionofsampleisrequired,pleasenotearea(s)ortakeatapeliftofarea. For Swab: • Forquantitativeculturereporting,theareaswabbedneedstobeenteredonthechainofcustody.

CulturableSurfaceSampling(BulkorSwab)

Benefits • Thesamplingmethodisinexpensiveandsurfacescanbequicklysampled. • Ausefultestforinitialsitesampling. • Specieslevelidentificationpossible. • Viabilityoffungiisdetermined.

Disadvantages • Areasoffungalgrowthareoftensmallandscattered,sotheymaynotallbepickedup.Multiplesamplelocations willhelpovercomethisproblem. • Healthproblemsrelatedtoindoormicrobialgrowtharegenerallycausedbytheinhalationofsubstantialnumbers ofairbornespores,sometimesoveralongperiodoftime.Thepresenceofbiologicalmaterialsonaparticular surface may not be a direct indication of what is in the air. • Thismethoddetectsonlyviablesporesandhyphaebutcannotdetectnonviableordifficulttoculturefungi.Itis advisabletocombinedirectexamsampleswithculturemethodsifknowingthepresenceofnon-viablefungiis important to your project. • Culturescannotdistinguishbetweenspores,hyphaeandotherfungalcells;theresultsarereportedascolony forming units.

Materials Required • Latex/nitrilegloves


Sampling Procedure

SwabSampling 1. Obtainsterile1mLButterfield’sSolutionswabtocollectand transportsamples(providedbyEMSL). 2. Wearinggloves,removeswabfrompackagingmaterial. 3. Removeplugfrommediatube. 4. Swab the desired area thoroughly, rolling the swab lightly back andforthoversamplingarea. 5. Insert the swab in the tube, firmly close cap, and label appropriately. 6. Forquantitativeculturereporting,theareaswabbedneedstobe entered on the chain of custody.

BulkSampling 1. Obtainsterilesamplingbags(Ziplock-type)tocollectandtransportsamples (providedatyourrequestbyEMSL). 2. Wearingglovesandusingcleantoolsremovearepresentativeareaofgrowthalongwiththebuilding material(sheetrock,wood,etc).1inchsquareissufficient. 3. Place bulk material into sampling bag and label the outside of the bag with sampling location or description.

Sample Shipment • CompleteanEMSLChainofCustody(COC),availableonourwebsite(www.emsl.com),detailingclient name and information, project name or number, sample #, and a description of the area. • Placesamplesinacoolerwithreusablefreezerpacks • Overnightshippingrecommended

Tech Tip: GeneralMediaRecommendationsForAirSamplingofCulturableFungiandBacteria

Fungal sampling in general, we recommend MEA. Malt Extract Agar is a general isolating media for culturingawide-spectrumoffungi.AgoodmediaformostofyourIAQprojects.Samplingforcellu-lose-degrading microfungi in water damaged buildings (e.g. Stachybotrys sp.) either Cellulose agar or Corn MealAgar.Bacterialsamplingingeneral,werecommendTSAorTSAw5%BloodSamplingforgramnegativebacteria,werecommendMacConkeyAgar.Allothersituations,theMicrobiologyDepartmentwillbehappytomakerecommendationbasedonyourindividualsamplingsituationorproject.

SterileSwab #8708301

FREE to EMSL Customers Call for Details


ERMI Dust Sampling

• TheERMI© is an acronym for EnvironmentalRelativeMoldiness Index. • ItwasdevelopedbyscientistsattheEnvironmentalProtectionAgency(EPA)toprovideastraightforward, objective,andstandardizedwaytoobtainresultsforindoorairqualityinvestigations. • TheEPAhasdevelopedanERMIrankingsystembasedondustsamplescollectedfromhomesacrossthe UnitedStates. • TheERMIhelpspredictthemoldinessofhomes.HomeswithhighERMIvalueshaveagreaterchanceof havingamoldproblemthenhomeswithalowERMI. • 36differentfungimakeuptheERMIandaredesignatedasGroupI(thosefoundinatypical,water damaged homes) and Group II (those commonly found in all homes).

Sampling Locations • Forresidentialsampling,EPArecommendstakingalivingroomandbedroomsampleasacomposite usingthesamevacuumdustcollectorforbothrooms.Otherareasshouldbesampledseparately. • IntheCommonLivingArea(familyroomorlivingroom),selectthesofa.Intheabsenceofasofa,select another commonly used chair. • Usingthetapemeasureandtherolloftape,markthecornersofa3footx6footrectangularsamplingarea onthefloorimmediatelyagainstthesofa.Placethelongsideoftherectangleagainstthelongsideofthe sofa. If the area cannot accommodate the recommended sampling area, adjust the dimensions accordingly Sampleatotalof18squarefeet. • Forthebedroom,selectthemostfrequentlyusedbedroom.Usingthetapemeasureandtherolloftape, markthecornersofa3footx6footrectangularsamplingareaonthefloorimmediatelyagainstthesideof thebedwheretheresidentismostlikelytogetinandoutofthebed.Ifpossibletrytohavetherectangular sampling area extend under the bed by 3 or 4 inches so that part of the sample goes under the bed. Place the long side of the rectangle against the long side of the bed. If the area cannot accommodate the recommendedsamplingarea,adjustthedimensionsaccordinglySampleatotalof18squarefeet. • Recordthefinalsamplingareayoumarkedwithtapeonthelinesnexttothebedroomyousampledonthis data sheet. • AreasotherthantheLivingRoom,FamilyRooms,orBedroomsmaybesampled.Ifyouchoosetosample otherareas,aseparatevacuumdustcollectorshouldbeusedforeacharea.Pleasecallthelabifyouhave anyquestions.

Sampling Procedure 1. Makesurethehoseattachmentisconnectedtoyourvacuumcleanerproperly.Turnonthevacuumcleanerto makesurethehoseattachmentispullingair,andthenturnoffthevacuumcleaner.Sometimesthedust collectiondevicewillnotfitcorrectlyontothehoseattachment.Ifthisisthecase,youmayuseducttapeor electrical tape to tape the dust collector to the hose attachment. 2. Usetheextensioncordasneededtoreachthemarkedareawiththevacuumhose. 3. Removebothcapsfromsamplingdevice.Placethecapsinalocationsoyoucanfindthemafterthetestiscompleted. Attachtheflat,roundendofthesamplerdevicetotheendofthehoseattachmentofyourvacuumcleaner. 4. Usetheslantedendofthesamplingdevicetocollectyoursample.Keeptheslantendofthesamplingdeviceflush with the surface to be sampled. 5. Turnonthevacuumcleanerandstartthewatchortimer.Starttimingthevacuumingprocedureusingthestopwatch. Try not to disturb the tape. Do not exceed the 5 minute sampling period. 6. Vacuumtheareacontainedwithintheductorelectricaltape.Dothisbypassingthesamplingdeviceoverslightly overlapping,imaginaryparallellineswithinthesamplingareaforabout5minutes.Ifnecessary,adjustyourrateof movementsothatatotalof5minutesisusedtovacuumtheentire18squarefootsamplingarea. 7. Movetothesecondroomandrepeatthevacuumingofthetargetarea.Afterthesamplingiscompleted,holdthe samplingdeviceupwardtowardtheceilingandturnoffthevacuumcleaner.Re-captheslantendofthesampling devicesoasnottolosethedustcollected. 8. Avoidvacuumingupanylargedebristhatisnotdust.Ifyouaccidentallysuckupthetape,pointthesamplingdevice towardtheceilingandturnoffthevacuumcleaner.Pickthetapeoutofthesamplingdevice.Turnthevacuumcleaner backonandreturntovacuumingthesamplingarea.Besuretoaccountforlostsampletimewhenyoudothissoyou get a total of 5 minutes of sampling time. 9. Separatethesamplingdevicefromthehoseofthevacuumcleanerandre-captheflatendofthedevice. 10. Afterthesmallcapsaresecuredonthedustsamplingdevice,makesurethereisdustinthesamplingcontainerbefore yousendittothelab.Ifnovisibledustisnoticed,repeatthesamplingprocedureinbothroomsindifferentlocations untilvisibledustispresentinthedevice. 11. If you lose the small caps, seal the openings completely and securely with duct or electrical tape.


Real-Time PCR

• EMSLoffersstate-of-the-artfungaldetectionandenumerationusingUSEPA-licensedPCR technology. Real-Time PCR is an excellent complement to your current sampling strategies. • Ifyoucurrentlyusesporetrapstorevealthegenusnamesoffungiinindoorenvironments,youcanalso submitreal-timePCRsamplestoquicklydeterminethespeciesnamesofanysignificantmoldsthat were found on the spore traps. • Ifyouuseculturetechniquestodeterminetheidentityofviable organisms, you can use real-time PCR to determine the species names of those organisms sooner.

Sampling Procedure

Air Samples 1. Obtaina3-piecePCRair/dust-samplingcassettefromEMSL. 2. Removetheupper(blue)andlower(red)plugsofthecassette. 3. Attachavacuumpumptothecassettethroughtheloweropening. 4. Sample as much air as desired through the upper opening. There is no upper limit to sampling time. 5. RecordtheVOLUMEofairsampledandshipcassettetoEMSL. Norefrigerationisneeded.

Dust Samples 1. Obtaina3-piecePCRair/dust-samplingcassettefromEMSL. 2. Removetheupper(blue)andlower(red)plugsofthecassette. 3. Attach a small piece of tubing to the upper opening. Cut a 45-degree angle at the end of the tubing. 4. Attachavacuumpumptothecassettethroughtheloweropening. 5. Begin collecting dust through the upper tubing. There is no upper limit to sampling time. 6. ShipthecassettetoEMSL.Norefrigerationisneeded.

SwabSamples 1. ObtainasterileTransPorterswab(Withoutbuffersolution, Product ID 8708301) from EMSL to collect sample. 2.Wearinggloves,removeswabfrompackagingmaterial. 3. Swab the desired area thoroughly, rolling the swab lightly back andforthoversamplingarea.Besuretorecordtheareasampled. 4. Insert the swab into the tube and firmly close cap. Ship the swab to EMSL.

BulkSamples 1. Obtainasterileswabandfollowdirectionsasdescribedaboveforswabs. 2. Alternatively,youmaysubmitapieceofthebulksampleforanalysis.

Water/Liquid Samples 1. All water samples should be taken in sterile sampling containers, which our laboratorycanprovideuponrequest. 2. Fill container with 100 mL of water, or as much water as possible and seal securelytopreventleakageduringtransport. 3. Transport sample using ice packs to EMSL

Culture Plates 1. Ifyouareinterestedinfastspeciationofyourculturable(viable)fungiinyoursamples,youmaycombine PCRwithcultureanalysis.Simplyaskthelaboratorytoholdyourculturesforfollow-upanalysisandrequest the desired PCR organism package.

PCR Analysis Individual Cassette

#8715309 List Price: $5.00

USD

E-Lite Pump # 8706004


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Allergen Sampling

Sampling Procedure

DustSamplingwithaDustCollectorKit (Kit available from EMSL, Product ID 8715600) 1. Insert the white filter tube securely into the dust collector, through the opening at the angled end. 2.Attachthedustcollectortothevacuumcleanerhoseortube. 3.Turnonthevacuumcleanerandvacuum4separateareasfor30secondseach,whereeachareaisabout 1/4squaremeter.Totalsamplingtimeis2minutesandtotalareasampledisabout1squaremeter. 4.Aminimumof100mgofdustisrequiredforallergenanalysis. 5.RemovethefiltertubecontainingthedustsampleandplaceitinasmallZiplocbagorequivalent. Placeentiredeviceinthebag. 6. Label the bag with your sample name or code and ship to EMSL Analytical, Inc. for allergen analysis.

DustSamplingwithaFilterCassette (Cassettes available from EMSL, Product ID 8715313) 1. 25mm0.45mMCEfiltercassettescanbeattachedtoavacuumpumpusingPVCtubing. 2.Aflowrateof10LPMissufficienttocollectdustintothecassette. 3.Werecommendusingatemplatetoestablishandstandardizesamplingareas(carpet,furniture,bedding,etc.). 4. Check clear window at inlet end of cassette to determine that an appreciable amount of dust has collected. 5.Aminimumof100mgofdustisrequiredforallergenanalysis.

IndoorAllergenAnalysesAvailable Test Code and Description: • M033LatexProteinbyELISA • M034CatDander(Feld1)byELISA • M035DogDanderDogDander(Canf1)byELISA • M036CockroachCockroach(Blag1)byELISA • M037DustMitesDustMites(Derp1&Derf1)byELISA • M038MouseMouse(Musm1)byELISA • M039RatRat(Ratn1)byELISA • M044IndoorAllergenGroupCat,Dog,Cockroach,andDustMitesbyELISA (EnzymeLinkedImmunosorbentAssay).

Note: Multiple allergens can be analyzed from a single dust sample.

EMSL Carpet Sampling Kit 25mm Kit, #8715314

List Price: $24.99USD

Allergen Sampler #8715600

List Price: $7.00USD


Water Sampling for Total Coliform, E. coli,&FecalColiform

Sample Collection • Allwatersamplesmustbetakeninapproved,sterilesamplingcontainers, whichourlaboratorycanprovideuponrequest.Treated(municipality/ distribution systems) water samples must be de-chlorinated with sodiumthio-sulfatewhichisprovidedinthe100mLlabissuedbottles. • Keepsamplingbottlecloseduntilitistobefilled.

Sampling Procedure

DistributionorSinkFaucetSample 1. Removeaeratorfromfaucet. 2. Rinse faucet with a bleach solution. 3. Flush line by turning tap fully on and letting it run for 2 to 3 minutes, or for a time sufficient to permit clearingtheserviceline. 4. Reducewaterflowtopermitfillingbottlewithoutsplashing. 5. Collect a 100mL sample to submit for analysis.

Well Samples 1. If a sample is taken from a well, fitted with a hand pump, pump the water for about 5 minutes before collecting sample. 2. Ifthewellisequippedwithamechanicalpump,collectsamplefromataponthedischarge. 3. Ifthereisnopumpingmachinery,collectsampledirectlyfromthewellbymeansofasterilizedbottle attached to a rope or stick of appropriate length. Fit a weight to the base of the bottle. Lower the bottle intothewell.Takecaretoavoidcontaminatingsamplebyanysurfacescum. 4. Afterthesamplehasbeencollected,tightenthelidsecurely.Usinganalcoholwipe,cleantheoutsideof the bottle.

Hold Times • Ifthesampleistakenforcompliancepurposesorforadischargepermit,FecalColiforms,Fecal Streptococcus, and Enterococci samples must be run within 6 hours of sample collection. • Ifthesampleistakenforcompliancepurposesorforadischargepermit,TotalColiforms,E. coli and HeterotrophicPlateCountsamplesneedtoberunwithin24hoursofsamplecollection. • Ifclientisawarethatholdtimesaregoingtobeexceededbutwouldlikeanalysisanyway,pleaseindicate this on the Chain of Custody. • Ifholdtimesareexceeded,thismayimpacttheresults.Thiswillbenotedonthereport.

ChainofCustody(COC) • ACOCmustcontainsamplingdate,timesampled,andsamplesource.Thisisneededtodeterminewhether or not samples are within hold times and correct analysis procedure. • IflookingforTotalColiformspleaseinformusifyouwouldlikePresence/Absence(Colilert)orEnumeration (MFT-Membranefiltrationtechnique). – Colilert test code: M017 – MFT test code: M018 • Drinkingwatersamplesforcomplianceorregulatorytestingneedtohavefreechlorinetestedinthefield and included on the chain of custody.

Turn Around Times • TotalColiform-Presence/Absence(Colilert)resultscanbegivenin24-48hours. • TotalColiformenumeration(MFT),FecalColiform,FecalStreptococcusandEnterococciwilltake 6-10 days.

Sample Shipping • Allsamplesmustbeshippedcold,notfrozenandshouldremainupright.Thebestmethodofshipmentfor thesesamplesistousefreezerpacks,notsodabottlesfilledwithfrozenwater,bagsoficeorlooseice. Samples that are not shipped properly can become contaminated or may be rejected by the lab.


SewageContaminationinBuildings

• Methodsavailabletotestforsewagecontaminationinbuildingsinclude: M117 – a culture-based method detecting Total Coliforms, E. coli, and Enterocci (48 hr TAT) M013 – a culture-based method detecting Total Coliforms, Fecal Coliforms, E. coli, and Fecal Streptococcus (6-10 day TAT) M095–anewPCR-basedmethoddetectingTotalBacteroides(24hrTATavailable)

• Theculture-basedmethodshavetheadvantageoflookingforonlylivingbacterialcontaminantswhichmay betheonlyonesofinterestifdiseaseisofconcern.Endotoxintestingcanalsobeofvaluetomeasurethetotal amountofGramnegativebacteriainthebuilding(seepage19).

• TheBacteroidestesthassomeadvantagesoverthetraditionalculture-basedtestsincluding: – Bacteroides is specific for fecal contamination from all sources – animals, birds, and human – Coliforms, E. coli, Fecal Streptococci, and Enterococci can grow in water, soil, sediments and on vegetationinuncontaminatedenvironments(E. coliisfoundinpristinetropicalenvironments). – Bacteroides willnotmultiplyintheenvironment. – Bacteroides outnumber coliforms by 1.000:1 and outnumber E. coli by10,000:1; therefore, the chance of finding Bacteroides is greatly enhanced. – Traditionalculture-basedtestsrelyonthepresenceoflivebacteria. Thesebacteriaoftenwillnotbeviableinindoorenvironments. THISMEANSTHATANEGATIVECOLIFORMorE. COLI RESULT DOESNOTMEANTHEABSENCEOFFECALCONTAMINATION. ThenewBacteroidestestovercomestheseculturelimitations.Thelaboratory candetectlive,nonviable,orviablebutnotculturablebacteria.

SamplingProcedureforallSewageContaminationTests 1. Obtainasterile1mLButterfield’sSolutionswab(EMSLProductID8708935) tocollectandtransportsamples(providedatyourrequestbyEMSL). 2. Withgloveson,removeswabfromsterilepackaging. 3. Carefullyunscrewcapofsamplingdevice–swabisattachedtothelidofthecap. 4. Gently press out excess solution from sampling swab by pressing the swab against the inside wall of the tube with a rolling motion. 5. Holdswabatanapproximate30°anglefromthesamplingsurface,takingcarenot to contaminate any part of the swab or the sampling site. 6. Usingfirm,evenpressuremovetheswabslowlyandthoroughlyoveranentire 4”x4”samplingarea,rewettingtheswabtipwiththeNeutralizingSolutionas needed.Firsthorizontally,thenvertically:

7. Aftersamplingiscomplete,carefullyputswabbackintovialandclosecaptight. 8. Label the sample using a permanent ink marker. 9. Backgroundsamplesinnon-fecallycontaminatedareasshouldbetaken.

Sample Shipping • Samplesthatwillbesubmittedforculture-basedanalysismustbeshippedovernightcold,notfrozen, within 24 hours from the sample collection. The best method of shipment for these samples is to use acoolerwithfreezerpacks,notbagsoficeorlooseice. • TherearenospecialshippingrequirementsorholdtimesforBacteroidessincetheywillnotmultiply outsideoftheintestine.However,donotstoresamplelongerthanamonthbeforeanalysis


Legionella Sampling

• Legionellaceaeareubiquitousinsurfacewater,groundwater,estuarineenvironments.Mostoutbreaksof disease due to Legionella are due to contaminated mist from cooling towers and potable water distribution systems.Exposureoccursfromcontaminated,aerosolizedwater. • Weofferfourdifferentculture-basedmethodsdependinguponthelevelofinformationyouneed.For more information about the different tests please go to www.legionellatesting.com. All culture tests require10-14daysbeforecompletion.

WealsoofferseveraldifferentPCRmethodsdependingupontheinformationyouneed.Moreinformation about these can also be found at www.legionellatesting.com. While the culture method is considered the “Gold Standard”, PCR is useful for ruling out possible Legionella contaminationintime-sensitiveprojects. PCRresultscanberequestedsame-day,24hrsand48hrsdependingonyourneeds.

WheretoLook ThisisalistofwatersystemsthathavebeendemonstratedtoharborLegionellabacteriaandhavebeenassociatedwithoutbreaks.TheirinvestigationshouldbeincludedinanyLegionella Risk Assessment program:

• Potablewatersystems • Coolingtowers • Aerosolgenerationduringthebiologicaltreatmentofsomeindustrial process wastewater streams. For example, pulp and paper manufacturing, foodandbeveragemanufacturingandpharmaceuticalmanufacturing. • Aerosolgenerationduringmunicipalwaterandwastewatertreatment • Raw,utilityorfirewater • Ornamentaloutdoorandindoorwaterfountainsandponds • Heatedswimmingpools • CPAPwater • Pottingsoil • Hottubs • Humidifiers • Metalworkingfluids • Medicaltherapyequipmentlikedialysisunits,nasogastrictubes,respiratoryequipmentandnebulizers, whirlpool baths • Commercialcarwashfacilitiesparticularlythoseusingrecycledwater • Supermarketvegetablemisters • Icemachinesinhotelsandhospitals • Outdoorbodymistersatballparksandamusementparks*

*Has not been associated with any cases of legionellosis.

Sampling Considerations • Watersamplesmustbecollectedinsterilevessels(availablefromEMSL). • A1Lwatersampleisrecommendedforpotablewaters(containingsodiumthiosulfate). • Lessthan1Lisacceptableifhigherlevelsofbacteriaaresuspected. • Chlorinatedwatermustbetreatedwithsodiumthiosulfate. • 100-250mListypicallysufficientfornon-potablewatersamples. • Inadditiontocollectingwatersamplesitisusefultocollectaswabofthebiofilmfoundinthewater supply pipes. • Samplesmustbetransportedtothelaboratorywithin24hrsincoolerswithfreezerpacks.


CoolingTowerSamplingLocations-SampleQuarterly • TowerMakeup • TowerSumpawayfrommakeup*(waterandswab) • InlettoHeatExchanger • OutletfromHeatExchanger • DistributionPack • TowerPack*(waterandswab)

PotableWaterSamplingLocations-SampleSemi-Annually • CityWaterMainEntryPoint • StorageTanks • HotWaterHeaterDrainPoint* • HotWaterReturn • LastPointonColdWater/FirstPointonHotWater • LastPointonHotWater*/FirstPointonColdWater • 10%ofSelectedOutlets(waterandswab)*

*Test routinely

Legionella Routine Sampling Locations • Testalllocationstoestablishabaseline • Testalllocationsduringanoutbreak

Sample Collection 1. Collect water samples from the or from the cooling tower basin water, hot water storage tanks, showers, water taps, etc. 2. If sampling for PCR analysis, collect 100 mL of water and another 1000 mL if potable water or 250 mL if non-potable water is sampled if culture results are also needed.

Sample Shipping • Labelsamplenumberonthebottleandrecordonthechainofcustody. • Useadistinctivenumberforeachsample.Completeallsampleinformationonasampledatasheetfor yourownrecord.Sendacopywiththesamplestothelaboratory.Youcandownloadacopyofthe Chain of Custody form on our websitewww.emsl.com. • Tightlycapthebottles.Makesurethatwaterdoesnotleakduringshippingandtransporting.Taping ofthebottlearoundthecapandneckwithelectricvinyltapeisrecommended.Placetapedbottlesin a clean plastic bag. • Placethesamplesininsulatedboxesoronfreezerpackstoprotectthemfromextremetemperature fluctuationsinthesummermonths.NEVER USE ICE OR DRY ICE. Stuff the box with foam chips tocushion,andsealtheboxsecurelyforshipping.Sendsamplesbyovernightexpresscarrier.Schedule samplingbetweenMondayandFridaysothatsamplescanbedeliveredtothelaboratorynolaterthan Saturday.


Methicillin-resistant Staphylococcus aureus(MRSA)Sampling

• Methicillin-resistantStaphylococcus aureus, (MRSA) is a type of bacteria that is resistant to treatment with antibiotics related to penicillin. This bacterium is generally spread through direct contact with the hands of a health care worker or person who is infected or carrying the organism. There are also community-acquiredinfectionsthathaveoccurredoutsidethehospitalenvironmentinplacessuchas gyms, schools, daycares, cruise ships, and elderly care facilities. • EMSLAnalytical,Inc.providesMRSAAnalysisofEnvironmental Samples (swab, bulk, or water). Please note, we are not able to accept clinical specimens for testing. • Cautionmustbetakenbywearinggloveswhencollectingsamplesfromcontaminateditems;puton cleanglovesimmediatelybeforecollection.Removeglovespromptlyafteruse,beforetouching non-contaminateditemsandenvironmentalsurfacesandwashhandsimmediatelytoavoidtransferof microorganismstootherenvironments.

Sampling Procedure

SwabSampling 1. Obtainasterile1mLButterfield’sSolutionswab(ProductID8708935)fromEMSLtocollectsamples for culture or a TransPorter swab (Product ID 8708301) for PCR testing. 2. Removeswabfrompackagingmaterialandswabthedesiredareathoroughly;rollingtheswablightly backandforthoversamplingarea.A4”x4”areaisrecommended.Inserttheswabintothetubeand firmly close the cap. Ship the swabs to EMSL.

BulkSamples 1. Wearinggloves,removeasmallpieceofthesuspectmaterial.Placepieceinsideacleansterile containerornewplasticbag(Ziplock). 2. Close the bag or cap the container and label appropriately.

Contact Plates 1. Storethecontactplateat2-8°C.Productislightandtemperaturesensitive;protectfromlight, excessiveheat,moisture,andfreezing. 2. Holdtheplatewiththumbandsecondfingeranduseindexfingertopressplatebottomfirmlyagainst theselectedtestsurface.Thesameamountofpressureshouldbeappliedforeverysample. 3. Donottwistormovetheplatelaterally.Lateralmovementspreadscontaminantsovertheagarsurface, thus making resolution of colonies

Sample Shipping • Allsamplesmustbeshippedovernightcold,notfrozen,within24hoursfromthesamplecollection. Thebestmethodofshipmentforthesesamplesistousefreezerpacks,notbagsoficeorlooseice. Samples that are not shipped properly could get contaminated and may be rejected by the lab. • EMSLoffersaculture-basedMRSAtestingmethod(CodeM133)andaPCR-basedmethod (Code M203) for faster turnaround times.


Endotoxin Sampling

• Endotoxinsarecompoundsthatarefoundinthecellwallsofgramnegativebacteria.Endotoxinscancause flu-likesymptomswheninhaledandmayaggravateasthma. • Highlevelshavebeenreportedfromavarietyofenvironments,suchasinrecycledwater,cottonmills, agriculture or wastewater treatment facilities, industrial wastewater mists, and contaminated room humidifiers. HighlevelsofEndotoxinsarefoundinanyrecycledfluid.

Summary of Procedures • Airsamplesarecollectedonendotoxin-freemembranefilters,storeddesiccatedwhereverpossible,orstored at 4°C. Endotoxin analysis can be done on air, water, or bulk samples.

Caution • Therearenoacceptedthresholdvalues,sorelativevaluecomparisonshavetobemadebetweenthesuspect area,andanon-suspectoneundersimilarenvironmentalconditions. • Sinceendotoxinlevelsmaybesignificantinoutsideair,thelattermaynotbesuitableforbackgroundlevels, especiallyinenvironmentsknowntohavehighlevelsofgram-negativebacteria(seeabove,1stparagraph). • Allsamplingutensilsmustbecertifiedendotoxin-free.PVCandpolypropylenematerialscannotbeused due to their affinity and binding with endotoxin.

Materials • FluidsandBulksamples:Collectinendotoxin-free(glassorpolystyrene)containers (EMSL Product # 8715711). • Forairsampling:low-flowpump,rotameterflexibletubingendotoxin-freepolystyrenecassettes(EMSL Product # 8715700) with 37mm diameter 0.4 μm polycarbonate membrane filters and AP40 glass-fiber backingpads.Thesemustbespeciallytreatedtoremoveallendotoxins. • Sterilesurgicalglovesshouldbewornforallsamples.

Sampling Procedure

Air Sampling 1. Cautionmustbetakentoavoidbreathingon,touching,orotherwise exposing the sampling containers to human contamination. 2. For indoor sampling, ensure that all doors and windows are closed. 3. Calibrate pump to 1.5 liters per minute 4. Usingglove,connectcassettetopumpwiththered-cappedendtoward the pump. Collect air samples for 4 hours per sample. 5. Disconnectcassetteandreplacetheprotectivecoveringbackonboththe inlet and outlet of the cassette, and wrap entire cassette in original packing and seal with tape.

LiquidorBulkSampling 1. Liquidsamplesmustbecollectedinendotoxinfreevialsonlywhichcan be purchased from the lab.

Sample Shipping • CompleteanEMSLChainofCustody(COC),availableonthewebsite(www.emsl.com),detailing client name and information, project name or number, sample #, and a description of the area. • ShipsamplestoEMSLAnalytical,Inc.assoonaspossible.Liquidsamplesshouldbeshipped onfreezerpacks.

Quality Control Recommendations • Multiplesamplesneedtobecollectedatasiteinordertocompareapossiblyaffectedareawith an unaffected one.

Endotoxin Individual Cassette

#8715700 List Price: $15.00

USD


SamplingintheFoodIndustry

FoodContactSurfaces Wash hands thoroughly with soap and water before and after sample collection.

Materials • Environmentalswabwith10mLBufferSolution(1persamplingsite) • PermanentInkMarker • Predeterminedsamplingsiteofapproximately 4” x 4” (~ 100 cm2), or for more intricate surface areas, anequivalentareaestimationora“perpart”sampleisacceptable. • Gloves(non-sterile)

Sampling Procedure 1. Withgloveson,removeswabfromsterilepackaging. 2. Carefullyunscrewcapofsamplingdevice–swabisattachedtothelidofthecap. 3. Gently press out excess solution from sampling swab by pressing the swab against the inside wall of the tube with a rolling motion. 4. Holdswabatanapproximate30°anglefromthesamplingsurface,takingcarenottocontaminateanypart of the swab or the sampling site. 5. Usingfirm,evenpressuremovetheswabslowlyandthoroughlyovertheentiresamplingarea,rewettingthe swabtipwiththeNeutralizingSolutionasneeded.Firsthorizontally,thenvertically:

6. Aftersamplingiscomplete,carefullyputswabbackintovialandclosecaptight. 7. Label the sample using a permanent ink marker. 8. Keep sample(s) at a refrigerated temperature (35° – 40° F) until it is submitted to the laboratory for analysis. Submit within 24 hours of sampling.


SamplingNon-FoodContactSurfaces

Wash hands thoroughly with soap and water before and after sample collection.

Materials • Whirl-PakBag,SterileSponge(pre-wetted),SterileGloves(Pre-packaged)1persamplingsite • Predeterminedsamplingsiteofapproximately 10” x 10” (100 in 2), for more intricate surface areas, anequivalentareaestimationora“perpart”sampleisacceptable. • PermanentInkMarker

Sampling Procedure 1. Separatetheglove/spongeportionfromthespongeWhirl-Pakattheperforation 2. Tear off the clear, perforated strip at the top of the Whirl-Pak bag 3. Putonthesterileglove. a. Removethesterileglovefromthepouchbythetopedgewithoutcontaminating (touching,breathingon,contactingetc.)theglove b.Removeaglovebyholdingitfromthewrist-sideopening.Avoidanycontactwiththeouter surfaceoftheglove.Insertcleanhandintoglove,takingcarenottopuncturetheglove. 4. Openthebagcontainingthesponge,wearingsterilegloves,beingcarefulnottotouch spongetoanythingbuttheglovedhand. 5. Withglovedhand,removespongefromthebag. 6. Usingfirm,evenpressuremovespongeslowlyandthoroughlyoversamplingarea. Firsthorizontally,thenvertically:

7. Return sponge back to Whirl-Pak bag taking care not to contaminate the spongeorthebagwiththeunglovedhand. 8. Close the bag by folding the top down three times and bending the wire ends overontothebag. 9. Label the sample using a permanent ink marker. 10. Keep sample(s) at a refrigerated temperature (35° - 40° F) until it is submitted to the laboratory for analysis.

Whirl-PakBags

2ozWrite-OnBags 500qty,#8708908


4ozWrite-OnBags 500qty,#8708909



Pathogens Min. aw Min. Max Max Min. Max. Oxygen pH pH %salt Temp.(°C) Temp. Requirement

Campylobacter 0.99 4.9-5.5 8.0 1.5-2.0 30-32 42-45 Micro-aerophilic jejuni

Clostridium 0.93-0.96 4.7 9.0 10.0 10.0 48-50 Anaerobe botulinum type A, and proteolytic B and F

Clostridium 0.93-0.96 4.7-4.8 9.0 4.5-6.0 3.0 45 Anaerobe botulinum type E, and nonproteolytic B and F

E. coli 0.93-0.95 3.6-4.7 9.5 7.5-8.0 0.6-3.0 45 Facultativeanaerobe

Listeria 0.92-0.95 4.8 9.6 8-12 0-2.0 45 Facultativeanaerobe monocytogenes

Salmonella spp. 0.92 4.0 9.0 8.0 5.0 46-47 Facultativeanaerobe

Shigella spp. 0.96 6.0 10.0 6.0 7.0 46 Facultativeanaerobe

Staphylococcus 0.85-0.86 4.0 10.0 18-20 5-6* 45-45 Facultativeanaerobe aureus

Vibrio cholerae 0.95 3.6-6.0 9.6 6-8 8.0 42-46 Facultativeanaerobe

Vibrio 0.94 4.8-5.0 9.6 8-10 5.0 43 Facultativeanaerobe parahaemolyticus

Vibrio vulnificus 0.95 6.3 9.0 6.0 13.0 44 Facultativeanaerobe

Yersinia 0.95-0.96 4.1-4.4 9.0 6-7 -1.0–1 44 Facultativeanaerobe enterocolitica

*Minimumtemperaturefortoxinformationis10°C Micro-aerophilic:requireslimitedlevelsofoxygen Anaerobe:requirestheabsenceofoxygen Facultativeanaerobe:growseitherwithorwithoutoxygen

LimitingConditionsforPathogenGrowth GuidanceprovidedbyEMSLAnalytical,Inc.


MicrobiologyCultureMediaInformation

EMSLAnalytical,Inc.isabletoprovidealargevarietyofsamplingmediatoourcustomers. Thefollowinginformationisdesignedtohelpaidintheselectionofculturemedia,providingthe mostrepresentativesampleoftheenvironmentpossible.Themedialistedbelowarenotexhaustive; ifyouhavespecificmediarequirements,pleaseletusknow.

Bacterial Media • TSA(TrypticSoyAgar) • TSAw/5%Blood • MAC(MacConkey) • HEAgar(HektonEnteric) • BCYEagar(BufferedCharcoalYeastAgar) • MSA(MannitolSaltAgar) • EMB(EosinMethyleneBlue) • BE(BileEsculinAgar) • ChocolateAgar • BHI(BrainHeartInfusion) • Xyloselysinedeoxycholateagar(XLDagar)

FungalMedia • MEA(MaltExtractAgar) • PDA(PotatoDextroseAgar) • CelluloseAgar • SAB(SabouraudDextroseAgar) • CornMealAgar • DG18(Dichloran-GlycerolAgar18) • BirdSeedAgar • MycobioticAgar

GeneralMediaRecommendationsforAirSamplingofCulturableBacteriaandFungi • Forfungalsampling,inmostcases,werecommendMEA. • ForfungalspeciesidentificationuseMEAandDG18plates • Forbacterialsampling,inmostcases,werecommendTSAorTSAw5%Blood. • WhensamplingspecificallyforGramnegativebacteria,werecommendMacConkeyAgar. • TheMicrobiologyDepartmentwillbehappytomakerecommendationsbasedonyourindividual sampling situation.


FungalCultureMedia

• MEA(MaltExtractAgar) GoldStandardmediaforthespeciationoffungi.

• PDA(PotatoDextroseAgar) General-purposemediausedfortheisolationoffungiandmolds.

• CelluloseAgar Multi-purposemicrobialmediausedtoisolateorforisolating hydrophilicfungimicrobesabletoutilizecomplexcarbohydrates. MostoftenrecommendedinIAQinvestigationsof Stachybotrys sp. contamination.

• CornMealAgar Generalpurposemediaforthecultivationoffungifromthe environment,commonlyusedinIAQinvestigationof Stachybotrys chartarum contamination.

• DG18(Dichloran-GlycerolAgar18) Mediacontaining18%glycerol,usedfortherecoveryofXerophilic yeastandmolds.Recommendedforrecoveryindrylocationssuch as dry powder storage and manufacturing areas.

• BirdSeedAgar FortheisolationofCryptococcus neoformans.

• MycobioticAgar FortheisolationofHistoplasmacapsulatum

Bacterial Culture Media

• TSA(TrypticSoyAgar) Generalpurposemediaforisolationandcultureofbacteria (grampositiveandgramnegative).

• TSAw/5%Blood Generalpurposemediaforcultivation,isolationanddetermination ofhemolyticactivityofbacteria.

• MAC(MacConkey) Forquantitativeproceduresfortheisolationofgramnegativebacteria and for the differentiation of those microorganisms based on fermentation of lactose.

• HEAgar(HektonEnteric) Fortheisolationanddifferentiationofgramnegativeenteric microorganisms.

• BCYEagar Fortheisolation,selectionanddifferentiationofLegionella sp. (BufferedCharcoalYeastAgar) formenvironmentalsamples.

• MSA(ManitolSaltAgar) FortheselectiveisolationanddifferentiationofStaphylococcus sp..

• EMB(EosinMethyleneBlue) Fortheisolationanddifferentiationofgramnegativebacteria.

• BE(BileEsculinAgar) FortheisolationandidentificationofGroupDStreptococcus sp.

• Xyloselysinedeoxycholateagar FortheisolationanddifferentiationofSalmonella sp.. (XLD agar)

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NewJersey – Piscataway (1,5) AsbestosandMoldTesting 1056SteltonRd.,Piscataway,NJ08854 Tel: 732-981-0550

NewJersey– Cinnaminson (1,2,3,4,5,6) Corporate Headquarters Asbestos,Lead,Mold,Bacteria,EnvironmentalChemistry, Industrial Hygiene, TO-15, Radiochemistry,PCR,FoodMicro,FoodChemistry,Materials Science and Pharmaceutical Testing 200Route130North,Cinnaminson,NJ08077 Tel: 800-220-3675

NewYork – Buffalo (1,5) AsbestosandMoldTesting 490RowleyRd.,Depew,NY14043 Tel: 888-260-5887

NewYork – Carle Place (1,3,4,5) Asbestos,LeadandMoldTesting 528MineolaAve.,CarlePlace,NY11514 Tel: 516-997-7251

NewYork – New York City (1,2,3,4,5) Asbestos,FoodMicro,Lead,MoldandBacteriaTesting 307West38thSt.,NewYork,NY10018 Tel: 866-448-3675

North Carolina – Charlotte (1,2) AsbestosandMoldTesting 376CromptonSt.,Unit71,Charlotte,NC28273 Tel: 704-525-2205

North Carolina – Kernersville (1,4) Asbestos,LeadandMoldTesting 706GralinSt.,Kernersville,NC27284 Tel: 336-992-1025

North Carolina – Raleigh (1,2,6) Asbestos,FoodMicroandMoldTesting 2500GatewayCentreBlvd.,Ste.600 Morrisville,NC27560 Tel: 919-465-3900

Ohio - Cincinnati (1,2,3) AsbestosandMoldTesting 4623WesleyAve.,SteF,Cincinnati,OH45212 Tel: 513-208-2226

Pennsylvania – Plymouth Meeting (1,6) AsbestosandMoldTesting 521 Plymouth Rd., Ste 107 Plymouth Meeting, PA 19462 Tel: 610-828-3102

South Carolina – Charleston Service Center 975 Morrison Dr., Bldg. C, Ste. A Charleston, SC 29403 Tel: 888-958-8170

Texas – Houston (1,2,5,6) Asbestos,FoodMicro,MoldandBacteriaTesting 8700 Jameel Road, Ste 190 Houston,TX77040 Tel: 713-686-3635

EMSL CANADA, Inc. - Ontario (1,6) Asbestos,Radon,LeadandMouldTesting 10FalconerDrive,Unit3 Mississauga,Ontario,L5N3L8 Tel: 289-997-4602

1 NVLAP Accredited* 2 AIHA EMLAPAccredited* 3 AIHAIHLAPAccredited* 4 AIHA ELLAP Accredited* 5 NELACAccredited* 6 A2LAAccredited*

*Certifications/Accreditations may be for specific Field of Testing and/or Lab Division; verify specific accreditation parameters and up-to-date status with your account representative.

26

LaboratoryLocations

Effective Date January 1, 2014©2014 by EMSL Analytical, Inc. All rights reserved.

LAB Services Include:Asbestos, Mold, Bacteria, Industrial Hygiene, Metals, Environmental Chemistry,

Food & Conumer Products, Allergens, PCR-Polymerase Chain Reaction (DNA),

Silica, Volatiles Scan, Formaldehyde by HPLC, Indoor Air Quality, Water,

Radon and Materials Testing.

www.emsl.comemail: [email protected]

UNITED STATES

C A N A D A

Midwest866.736.4824877.435.5227

North East 800.220.3675

South East888.958.8170877.476.5866

South Central866.318.3920

Toronto289.997.4602

West Coast888.455.3675866.798.1089

Headquarters

EMSL Laboratory Location

EMSL Service Center Location

LA Testing Laboratory Location

EMSL Analytical Locations

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