environmental microbiological monitoring -...

EnvironmentalMicrobiological Monitoring

Bacteria, Fungi, Virus(Part I)

Roberto Ligugnana 2010

International pbi S.p.A.

International pbi Spa

The Company


The people


The Production


The Logistic


The Museum


The Cross CountrySki field


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


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


Air, Surface, Hands

PharmaceuticalAgro-FoodDairyCateringVeterinaryCosmeticsBeveragesDisinfectants

HospitalsVaccinsPublic HealthIndoor Air QualityWasteWater PlantsHeating Ventilation Air ConditioningBiotechnology


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


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


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


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


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


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


Why hygiene monitoring in the cosmetic industry?

Check the effectiveness of cleaning and disinfection programs

Improve hygiene education of staff

Improve Company image


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


Why hygiene monitoring in the veterinary field?

Regular control of disinfectant effectiveness

Control of animal environment for staff health


Why hygiene monitoring in wastewater treatment plants?

Regular control of bioaerosol on plant premises to monitor staff and neighbours health


Why hygiene monitoring in vaccin production facilities?

Monitoring the bioaerosol of the environment for staff health safety


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


Which part of the environment should be monitored for hygiene control?

1. AIR

2. SURFACE

3. HANDS


AIR/BIOAEROSOL

The particules carrying micro-organisms can spread and deposit everywhere with consequent microbiological contamination.


SURFACE

Surfaces contaminated by micro-organisms

may transfer the contamination to other

materials that are deposited on them or flow

through them.


HANDS

Microbiological cross contamination between staff and staff/surfaces can spread micro-organisms everywhere creating problems.


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


“What do you need for Surface Monitoring?

“Rodac-Weight” surface sampler

“Surfair” Contact Plate (Rodac)

Wall Poster for protocol and micro-organisms identification


What do you need for Hands monitoring?

“Maxi” Contact PlatesWall poster for protocol and micro-organisms identification


AIR SAS Air Sampler

SAS Surface Air System

Your partner in environmentalmicrobiology and virology


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


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.


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)



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)



SAS SUPER 100 /180Clean room air sampler: 100 /180 lts/minIdeal for clean roomsin pharmaceuticalindustries and high risk environments


QUICK SAMPLING2 samplings of 1000 liters (=1m3)

in about 3 minutes

REPRESENTATIVEContemporary sampling on 2

different media

DUO-SAS -360


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


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)


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


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.


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


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.


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


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.


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


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


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


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


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


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.


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

http://www.internationalpbi.it/en

http://www.internationalpbi.it/en

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