Environmental Microbiological Monitoring Bacteria, Fungi, Virus (Part I) Roberto Ligugnana 2010 International pbi S.p.A.
EnvironmentalMicrobiological Monitoring
Bacteria, Fungi, Virus(Part I)
Roberto Ligugnana 2010
International pbi S.p.A.
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The Company
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The people
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The Production
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The Logistic
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The Museum
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The Cross CountrySki field
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The literature:Produrre Bene e IgienicamenteThe pbi laboratoryGazetteThe EuropeLaboratory Times
Content
1-4 Introduction5-16 Why Hygiene Monitoring?17-20 Air, Surface, Hands21-26 What do you need?27-43 Air Sampling
44-47 Surface Sampling48-50 Hands Sampling51-54 Recording, Poster55-60 EU GMP in Clean Room
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The SAS air sampler in space
The SAS air sampler is on board of the space laboratory to monitor the Indoor Air Quality for scientists good health
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Air, Surface, Hands
PharmaceuticalAgro-FoodDairyCateringVeterinaryCosmeticsBeveragesDisinfectants
HospitalsVaccinsPublic HealthIndoor Air QualityWasteWater PlantsHeating Ventilation Air ConditioningBiotechnology
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Why hygiene monitoring for Public Health?
Improve hygienic conditions of public environment (schools, railways, etc.)Assure good hygiene conditions in restaurants, refectories, snack bars, etc.Check the effectiveness of disinfection programsAvoid biohazards to staff involved in high risk jobsImprove hygiene education of staff
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Why hygiene monitoring in the agro-food field?
Extend shelf-life of foodImprove Hygiene education of staffCheck the effectiveness of disinfection programsImprove image of the company for end-users, customers, suppliersImplementation of HACCP System
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Why Hygiene monitoring in the pharmaceutical field?
Regular and programmed control of clean rooms according to FDA/Pharmacopeia
Improve the hygiene education of the staff
Check the effectiveness of disinfection programs
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Why hygiene monitoring in Hospitals?
Regular control of operating theatres, high risk areas, nurseriesCheck the effectiveness of disinfection programsHygiene education of staffReduce risk of staff contaminationProvide the best service to high risk patients
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Why hygiene monitoring in the microbiological laboratory?
Control of laminar flow bench
Control of laboratory environment for staff health
Good co-operation with the production Department in agro-food-dairy industry
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Why hygiene monitoring in buildings, houses?
Control the Indoor Air QualityAvoid Sick Building SyndromeLocate airborne infections (legionella)Check the effectiveness of cleaning and disinfection programsScheduled monitoring of air ducts in HVAC
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Why hygiene monitoring in the cosmetic industry?
Check the effectiveness of cleaning and disinfection programs
Improve hygiene education of staff
Improve Company image
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Why hygiene monitoring in the disinfectant industry?
Regular control of disinfectant effectiveness
Evaluate the correct use of disinfectants
Show the customers the performances of the disinfectants
Report on new disinfectants or competitor’s disinfectants
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Why hygiene monitoring in the veterinary field?
Regular control of disinfectant effectiveness
Control of animal environment for staff health
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Why hygiene monitoring in wastewater treatment plants?
Regular control of bioaerosol on plant premises to monitor staff and neighbours health
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Why hygiene monitoring in vaccin production facilities?
Monitoring the bioaerosol of the environment for staff health safety
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Why hygiene monitoring in High School and University?
To show and educate the student about the presence of micro-organisms in the air
Microbiological exercises and experiments
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Which part of the environment should be monitored for hygiene control?
1. AIR
2. SURFACE
3. HANDS
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AIR/BIOAEROSOL
The particules carrying micro-organisms can spread and deposit everywhere with consequent microbiological contamination.
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SURFACE
Surfaces contaminated by micro-organisms
may transfer the contamination to other
materials that are deposited on them or flow
through them.
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HANDS
Microbiological cross contamination between staff and staff/surfaces can spread micro-organisms everywhere creating problems.
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What do you need for Air Monitoring?
“Surface Air System” SAS air sampler“Surfair” Contact Plates (RODAC) or standard 90 mm Petri dish“SAS software” for time, locations and operator recordingWall Poster for protocol and micro-organismidentification
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“What do you need for Surface Monitoring?
“Rodac-Weight” surface sampler
“Surfair” Contact Plate (Rodac)
Wall Poster for protocol and micro-organisms identification
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What do you need for Hands monitoring?
“Maxi” Contact PlatesWall poster for protocol and micro-organisms identification
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AIR SAS Air Sampler
SAS Surface Air System
Your partner in environmentalmicrobiology and virology
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AIR Purpose of SASSurface Air System air samplers
To use a simple and inexpensive ContactPlate or Petri dishTo produce quantifiable results in relation to the sampledair volumeTo allow the operatorto use different agar ofhis choice
To introduce active air samplingTo establish data on the microbial level in different environmentsTo improve hygiene standards with regular monitoring and recording of results
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AIR Principle of SASSurface Air System air samplers
A known volume ofair is tested.Bioaerosol isaspirated through a detachable cover with multiple holes.The plate is thenincubated.
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AIR Principle of SAS Surface Air System air samplers
The colonies are then visible to the naked eye and can be counted. The results are reportedas CFU/m3 of air.
AIRDifferent models of SAS air samplers
SAS SUPER 180High volume air sampler: 180 lts/minIdeal for applicationswhere a large volume of air should beaspirated rapidly(1000 lts of air in about 6 minutes)
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AIRDifferent models of SAS air samplers
SAS SUPER 100 Medium volume air sampler:100 lts/min
Ideal for applicationswhere portability is a priority (1000 lts of air in about 10 minutes)
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AIRDifferent models of SAS air samplers
SAS SUPER 100 /180Clean room air sampler: 100 /180 lts/minIdeal for clean roomsin pharmaceuticalindustries and high risk environments
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QUICK SAMPLING2 samplings of 1000 liters (=1m3)
in about 3 minutes
REPRESENTATIVEContemporary sampling on 2
different media
DUO-SAS -360
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SAS Isolator
The air sampler tobe used in isolators
SAS Multi Isolator
Several aspirationheads with just a single comand unit
Pinocchio Super II
Microbiologicalsampler forcompressed air
SAS Super IAQ
The air sampler forIndoor Air Quality
SAS PCR
The air sampler tocollect pathogenmicro-organismsand subsequentidentification byreal time PCR
SAS Super Network
Several air samplers positionedat differentlocations withinClean Roompremises
Different type ofaspiration heads
Sterilizable head:AISI stainless steel orAluminium
Different type ofaspiration heads
Sterile head:Plastic “Daily-Head” withcertificate ofsterilization(specific for CleanRoom)
“Surfair” and “Maxi” Contact Plates
The Contact Plates have a convex surface of nutrient agar over the edge of the plate that can directly touch the surface to be tested to take a microbiological picture of it
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Empty “Surfair” Contact Plates
The Contact Plates are available with or without media.Each empty “Surfair” Contact plate should be filled with 16-18 ml of media.The “Maxi” should be filled with 28 ml of media
After cooling the Contact plates shouldbe dried for a fewminutes under laminar flow and then stored at controlled temperature (2-8 °C)
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Ready Poured “Surfair” Contact Plates
Ready poured Contactplates should be storedat roomtemperature(10-25°C)They are available in different typesPacked in single blister (20/pack) for a better storage whenthe box has been open.
Clean-room packing for high risk environment(3 protective layers)Media with added neutralizer for possible disinfectant present on the tested surface that can inhibit organism growth
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Microbiological Media
Total Bacterial Count: Tryptic Soy Agar
Moulds and Yeasts Count: Sabouraud Dextrose Agar
Agar Contact blister
Individuallypackaged ready touseTriple packaging for Clean Room
Agar Petri
Ready to use tripplebagged Petri platesfor use in CleanRoom
AIR “SAS” References
“SAS” meets the requirements as for FDA 1987 Guide on Sterile Drug Products.
“SAS” is referenced in ACGIH Guide-line forbioaerosol in Indoor Environment.
“SAS” is included in a protocol considered byASTM Committee D22.05.06.
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USP 23-NF 18 - 8th Supplement (May 98) -Microbiological Evaluation of Clean Roomsand other Controlled Environments.EU Gude fo GMP - Manufacture of SterileMedicinal Products Control of Medicines andInspectionCEN/TC 243 Norms for Clean RoomsTechnology
AIR “SAS” References
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AIRSAS air sampler: method of use
1.Remove cover of sampler
2.Insert closed filled Contact Plate and remove plate lid
3.Replace cover of sampler
4.Select air flow and switch on the unit
5.Remove sampler cover at the end of cycle
6.Remove covered Contact Plate
7.Incubate8.Count CFU, record
results, refer to 1000 lts/air, interpret results.
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AIRSAS SUPER 100 : method of use
SAS SUPER 100 meets the needs of clean room activityThe transparent, portable, light, autoclavable case allows an easy transfer into the clean areas, eliminating contamination risks
The head of the instrument made in AISI 316 polishedstainless steel guarantees efficientcleaning, disinfectionand less metal particleemissionFollow the “SAS Super 100” instructions
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AIRSuggested Procedures for Air Sampling
The next slide gives suggested regimes whichmay be adopted.In environments with constant characteristics itis easier to provide “Acceptability Levels” tobe used for routine work.It is not possible to set the same standards forall applications and the “A. L.” will varyconsiderably in different environments.
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AIRSuggested regime for air sampling
Hospital High risk Every 15 daysLow risk Every 30 days
Pharmac. Cl.Room Every dayFood Product. Every weekDairy Product. Every weekHealthDp Commun. Every 90 daysBactLab LaminarF Every 120 days
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Suggested Frequency of Sampling On the Basis of Criticality of Controlled Environment by U.S.P
Class 100 roomAdjacent areas to Class 100 roomOther support areas (Class 100,000)Potential product contact areas
Each operating shift
Daily
Twice / week
Twice / week
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AIR Calculation of results after air sampling
The number of organisms counted on a Contact Plate must be corrected for the statistical possibility of multiple particles passing through the same hole(use the Correction Table)
The final result in CFU/m3 of air is obtained using the formula:CFU/plate x1000
x=--------------------Volume of air
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AIRSuggested Interpretation in Clean Rooms
It is necessary to comply with figures indicated by International Organisations (NASA,Pharmacopeia, etc.)
Example: Nasa Standard NHB 5340-2Class 100 less than 0,1 CFU/ft3Class 10000 less than 0,5 CFU/ft3Class100000 less than 2,5 CFU/ft3
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EU for GMP Guide
Recommended limits for Clean Room
Grade Air Sampler Contact Plate GloveImprint
A <1 CFU/1m3 <1 CFU/plate <1 CFU/gloveB 10 CFU/1m3 5 CFU/plate 5 CFU/gloveC 100 CFU/1m3 25 CFU/plate -D 200 CFU/1m3 50 CFU/plate -
WHO Suggestions for HospitalType of environment Micro-organisms per 1000
litres of airEnvironment classification
Operating theatreSpecial roomsSpecial Laboratories
Up to 10 CFU/1000 litres of air I. Level - Very low environmental bacterial count
Operating theatre of first aidOperating theatre adiacent roomsSala partoNido per lattantiUnità centrale di sterilizzazione (“zona pulita”)
Up to 200 CFU/1000 litres of air
II. Level - Low environmental bacterial count
Patient room Over 200-250 CFU/1000 litres of air
III. Level
AIR Suggested interpretation in normal environments
It is recommended that at least 30 samples
are taken. The results are plotted on graph
paper and the mean value is calculated. The
plotted mean value will indicate the “goal”
to be lowered during the next air monitoring
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Normal Environment
INSERIRE GRAFICO COME DA AGGIORNAMENTO PORTALE N.15
AIR Validation/Calibration
Air samplers require a periodic validation. The frequency of this control depends on the frequencyof use.The official validation must refer to officialdocuments for the referability of protocol and results.For official validation “SAS” is validated with the “Polutecnic” unit with a certified anemometer.
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Calibration / Validation
The air samplersare monitored fortheir regular functionment usingthe “Pyramid” System
Calbration / Validation
The air samplersshould be officiallyvalidated every6/12 months using“”Politecnic”, according to FDA inspectors request
SAS in WEB
Several ApplicationNotes may bedownloaded fromthe www.internationalpbi.it/en portal