© PharmOut 20171

Cleanroom Contamination

including Mould/Fungal

contamination

Brett Cole. Biosafety Pty Ltd

Tuesday 25th July 2017

© PharmOut 20172

1. Introduction

2. What is Contamination?

3. What is Mould?

4. Sources of Mould/Fungal contamination

5. How to minimise mould/fungal contamination

6. Cleanroom decontamination

7. Planning, Execution and Validation

8. The various common and emerging technologies used for manual

and rapid decontamination and sterilization

9. Case Studies

10. Conclusion

11. Questions

Cleanroom Contamination including Mould/Fungal contamination:

© PharmOut 20173

• Bachelor of Science with Honours Degree – Microbiology (Monash University)

• Masters Degree in Occupation Hygiene and Toxicology (Edith Cowan University)

• Over 15 Years experience in Infection, Waste and Contamination Control (Healthcare, Pharmaceutical and Laboratory)

• IICRC Certified for Water Damage and Mould/Microbial Remediation • Committee Member of ABSANZ Regulatory Committee (OGTR, DAFF

and AS/NZS)• Internationally Certified Biorisk and Biowaste Management

Professional (IFBA)• Member of Australia Institute of Occupational Hygiene (MAIOH)• Member of Australian Standards Committees (CH-029) Safety in

Laboratories Standards• Licensed Fumigation Company (Dept of Health and Human Services)• Member of ANZLAA, ABSANZ, SCRIA, ISPE and ACPIC• 2017 Australian Restoration Awards – Most Innovative Specialised

Restoration Project – Microbial Infestation Restoration of a Medical Research Facility

Professional Experience-

© PharmOut 20174

What is contamination?Contamination is the introduction of unwanted material or substance into a process, person or product.

It is particularly critical in controlled environments such as

➢ Food

➢ Pharmaceutical

➢ Healthcare

➢ Life Science

How does a facility get contaminated?

How does your product get contaminated?

Contamination can occur at any step in the process—during production,

processing, distribution, transportation, preparation, etc.

© PharmOut 20175

What is mould?

• From the Kingdom Fungi

• Not Animal or Plant

• Form of multicellular thread-like structures called hyphae.

• Some Fungi exist as single cells are called yeasts.

• Can cause disease or food/product spoilage, others play an

important role in biodegradation or in the production of various

foods, beverages, antibiotics and enzymes

• Can reproduce sexually or asexually via spore release

• Require high relative humidity to exist (>60% RH)

• Cannot produce their own energy

• Require organic material to feed on

• Can release airborne Mycotoxins

© PharmOut 20176

What is mould?

• Some are good

• Some are bad

© PharmOut 20177

Sources of mould and contamination in cleanrooms.

• Primary source of mould/microbial contamination into cleanrooms is

via personnel/humans

• Poor gowning procedures

• Poor Ingress and egress/access control

• Poor Heating, Ventilation and Air Conditioning (HVAC) maintenance

and hygiene

• Equipment and raw material introduction into clean rooms

• Cellulose material such as cardboard, paper and wooden pallets

• Contaminated Packaging

• Flooding introducing spores from external environments

• Poor or improper decontamination processes

• Inappropriate cleanroom construction materials

• Leaks in cleanroom structure

© PharmOut 20178

How to minimise mould/fungal growth?

• Facilities should be constructed of nonporous material that do not

absorb water and promote fungal growth (21 CFR 211.42)

• If floods occur, there is a need to react quickly to water damage

within a facility aggressively and fully dry or replace damaged walls

and ceilings within 72 hours to prevent mould growth.

• Temperature and humidity controls must be in place to discourage

fungal growth, that is, RH < 60% RH.

• Minimise cellulosic materials such as cardboard and wooden pallets

from GMP areas as they can become wet and support fungal

growth.

• Cleaning and disinfection procedures must be in place to prevent

people and mobile equipment introducing fungi into facilities.

© PharmOut 20179

Decontamination

© PharmOut 201710

Why Decontaminate?

➢To create a Sterile baseline

➢Renovation (Before or after)

➢Between Population / Production Batches

➢Commissioning

➢De-Commissioning

➢Contamination/Infection

➢Preventative Maintenance

➢Regulatory compliance

➢GMP/Best Practice

➢Other…

© PharmOut 201711

What makes for a good decontamination?

➢All Decontamination methods can work based on

the following:

➢ Must reach ALL surfaces for a prescribed

amount of time, which means you must have:

1. Good and Complete Distribution

2. Thorough and Total Penetration

3. Sufficient Contact Time

4. At specified concentration

5. At required Environmental conditions

(temp/RH)

➢Any decontamination method requires a complete

and thorough distribution of the sterilant or high

level liquid disinfectant to get an effective

decontamination or kill

© PharmOut 201712

Planning for decontamination:

• Define purpose and scope: rooms, equipment, HVAC

• Identify the players, area production managers,

• OH&S, facilities engineering, QC validation, security, emergency

services, other facility users

• Establish responsibilities (SWMS, SOP, Who’s who)

• Select decontaminating agent (Best fit-for-purpose)

• Establish the schedule and ordering

• Write SOPs, fumigation management plan (AS 2467 24 hour

notification) where required

• Define validation: Microbial Testing, full PQ, target dosage

• Hand back procedure

References: Harris (2010)B & V Testing, INC.

AS 2467 (2008) General Requirements for Fumigation

© PharmOut 201713

Microbial Resistance:

© PharmOut 201714

Sterilizers (Sporicides): Used to destroy or eliminate all forms of microbial life including fungi, viruses, and all forms of bacteria and their spores. Spores are considered to be the most difficult form of microorganism to destroy. Therefore, EPA considers the term Sporicidal to be synonymous with "Sterilizer.“ (Log 6 Reduction)

Disinfectants: Used on hard inanimate surfaces and objects to destroy or irreversibly inactivate infectious fungi and bacteria but NOT necessarily their spores. Disinfectant products are divided into two major types: hospital and general use. (Log 4 Reduction)

Sanitizers: Used to reduce, but not necessarily eliminate, microorganisms from the inanimate environment to levels considered safe as determined by public health codes or regulations. (Log 2 Reduction)

Antiseptics and Germicides: Used to prevent infection and decay by inhibiting the growth of microorganisms. Because these products are used in or on living humans or animals, they are considered drugs and are thus approved and regulated by the Food and Drug Administration (FDA).

http://www.epa.gov/oppad001/ad_info.htm

Types of Antimicrobial applications:

© PharmOut 201715

Types of Antimicrobial applications:Liquid Treatments:

• For mould/fungal decontamination, the antimicrobial must be sporicidal and fungicidal

• Sterile cleaning equipment must be used in conjunction with the appropriate antimicrobial of choice.

© PharmOut 201716

Rapid Decontamination Systems:

• Formaldehyde

• Ozone (Cold Plasma/UV Generated)

• Vapourised Hydrogen Peroxide

• Ionised Hydrogen Peroxide

• Chlorine dioxide

• Nitrogen dioxide (small volumes)• All of the above fumigants are sporicidal and fungicidal

© PharmOut 201717

Comparisons between technologies:

Key

➢H2O2-1 = Wet VHP Process

➢H2O2-2 = Fogging Process

➢H2O2-3 = Dry VHP Process

© PharmOut 201718

Case Studies

© PharmOut 201719

19

Documented Mould Contamination Case Studies:

2015 - Mould in a anti-tuberculosis manufacturing facility in India. WHO issued

a Notice of Concern (NOC) for black mould found in GMP facility. Product recall

and shortage of supply.

http://www.in-pharmatechnologist.com/Regulatory-Safety/Black-mould-and-

rusty-equipment-lands-Indian-TB-drugmaker-WHO-warning

2012 – New England Compounding Centre shut down after 64 meningitis

related deaths from fungal contamination of products.

https://www.cemag.us/blog/2017/01/pharmacists-fatal-meningitis-outbreak-

facing-trial

© PharmOut 201720

20

Case Study: BioSperix Cell Isolator:

Tissue Culture Facility with mould in the isolator. Hard to reach with manual

cleaning

© PharmOut 201721

21

Case Study: BioSperix Cell Isolator:

© PharmOut 201722

22

Case Study: Cell Culture Laboratory with Mould Contamination caused by

Mould biofilm in the ceiling space:

Source removal and application of antimicrobial

© PharmOut 201723

23

Case Study: Cell Culture Laboratory with Mould Contamination caused by

Mould biofilm in the ceiling space:

Source removal and application of antimicrobial

© PharmOut 201724

24

Case Study: Facility Decontamination – Animal Facility – Pinworm Outbreak

Facility with single stage physical clean and pinworm egg Inactivation using

Chlorine dioxide gas

Total Project Duration = 7 days (6 day clean, 1 day gas)

© PharmOut 201725

25

Case Study: Facility Decontamination – Animal Facility – Pinworm Outbreak

© PharmOut 201726

Summary:

• Identify possible sources or routes of contamination• Control environmental factors such as relative humidity to reduce

the chance of fungal growth• Control the introduction of organic or cellulose based materials

into your cleanroom• Have GMP processes in place to correctly measure and mitigate

the opportunity for fungal contamination, ie; cleaning regimes, HVAC hygiene etc

• Understand the physical properties and behaviour of your decontaminant of choice to ensure efficacy and safety and is a sporicide and fungicide

• Different decontaminants may be appropriate for different applications

© PharmOut 201727

Good References:

• IICRC S500 Guideline for Water Damage• IICRC S520/R520 Guideline for Mould Remediation• http://www.ivtnetwork.com/article/understanding-preventing-and-

remediating-mold-cleanrooms

© PharmOut 201728

© PharmOut 201729

ANY QUESTIONS?

Thank you for your time!

Please feel free to email me at brett@biosafety.com.au