Risk-based Environmental

Monitoring Program
 2

Presented by
Ziva Abraham
President
Microrite, Inc.
5019 New Trier Avenue
San Jose, CA 95136
Phone: 408-445-0507 Fax: 408-445-1236
Email: [email protected] www.microrite.com
About the Presenter 3

Ziva Abraham is the President and Founder of Microrite, Inc., a California based consulting
firm providing consulting and training services to pharmaceuticals, biotechnology, medical
devices and in vitro diagnostics in the areas of quality assurance, quality control,
microbiology, and validation.
Ziva has over 35 years of academic, research, clinical and industrial experience in
microbiology, and quality assurance. Ziva has received her Master’s Degree in microbiology
with a focus on Mycology and has conducted research on developing microbial Insecticides
using entomogenous bacteria and fungi for her Ph.D. degree.
Her career also includes founding and managing clinical laboratories for Maccabi Medical in
Israel. She has trained personnel from various industries in microbiology techniques and
methods. She uses her extensive experience to teach why assessing risk of microbial
contamination should be in the forefront of any company that has products for
human/veterinary use.
Her experience in clinical laboratories has provided her with the framework to understand
the effects of microbial contamination in products from a patient safety perspective.
Main Message 4

Environmental Monitoring is not an isolated activity; to develop a risk

based environmental monitoring program, risk across the lifecycle of
the facility, process, personnel and operations must be assessed utilizing
pertinent knowledge!
Quality Risk Management (QRM): concept:
• Risk Management is NOT an Exercise!!!
• Real-world risk must be considered and addressed
• Risk Management should be:
o Pragmatic
o Scientifically Sound
o Utilize Current Technology and Current Best Practices
Learn from experience and from the mistakes of others!
No single SME can be an expert in all aspects of Risk Assessment!
Current Regulatory Expectations 5

GMP Annex 1 Manufacture of

Sterile Medicinal Products:
2008, Drafts 2017 and 2020

FDA
FDA: Guidance for Industry Sterile Drug Products
Produced by Aseptic Processing -Current Good
Manufacturing Practice:2004
Why a Risk Based Environmental Monitoring Program? 6

These risk assessments should be re-evaluated at defined intervals in

order to confirm the effectiveness of the site’s environmental monitoring
program, and they should be considered in the overall context of the
trend analysis and the contamination control strategy for the site.
• Facilities age
• Barrier and cleanroom design and integration may not be optimal
• Leaks in the room occur
• Membranes tear
• Filters get loaded
• Gowns wear
• Personnel leave
Risk Assessment?? 7
FDA 483 Observation 8

Poor Aseptic Behavior

0ur investigator observed multiple poor aseptic practices during the set-up and
filling of (b)(4) batch (b)(4).

For example, during the aseptic filling of vials, an operator used restricted access
barrier system (RABS) (b)(4) to remove a jammed stopper by reaching over
exposed sterile stoppers in the stopper bowl.

The RABS (b)(4) disrupted the unidirectional airflow over the stopper bowl,
creating a risk for microbial contamination. After the operator removed the
jammed stopper, the filling line was restarted, but the affected stoppers were not
cleared.
FDA 483 Observation 9

Our inspection also revealed poor aseptic processing operation behaviors. In response to this
letter, provide:
Your plan to assure appropriate aseptic practices and cleanroom behavior during production.
Include specific steps to ensure routine supervisory oversight for all production batches. Also
describe the frequency of quality assurance oversight during aseptic processing and other
operations.
Comprehensive identification of all contamination hazards with respect to your aseptic
processes, equipment, and facilities.
Provide a risk assessment that covers all human interactions with the ISO 5 area, equipment
placement and ergonomics, air quality in the ISO 5 area and surrounding room, facility
layout, personnel flow, and material flow.
Also include a detailed CAPA plan, with timelines, to address the findings of the
contamination hazards risk assessment.
Understanding Contamination Sources and Risk 10

Without knowing the risk, risk cannot be monitored!

• Using rating scales that are neither specific nor appropriate to a given situation
• Perceived risk instead of understanding real-world risk
• Not acknowledging uncertainty or missing important information
• Neglecting to keep risk assessments current (reflecting current regulatory
expectations)
• Performing paper-based risk assessments without understanding systems,
workflows and all possible sources of contamination
• More about the presentation rather than the risk
• One person performs the entire risk assessment?
FDA 483 Observation 11

Aseptic garments worn in the filling area were also non-integral.

“We observed 7 of (b)(4) sterile gowns with tears or holes; 8 of (b)(4) had loose
threads. We observed 2 of (b)(4) sterile hoods with tears or holes; 12
of (b)(4) had loose threads. We observed 8 of (b)(4) sterile booties with tears or
holes; 11 of (b)(4) had loose threads.”

Procedure "Handling of Aseptic Area Garments" required production personnel

to examine the garments for tears, holes, and loose threads, but our investigator
found that these checks were not being performed.

https://www.microrite.com/wp-content/uploads/2020/01/MICROBIOLOGICAL-FAILURES-DUE-TO-HUMAN-
BORNE-CONTAMINATION-CLEANROOM-GARMENT-AND-MANAGEMENT-GAPS-2020v1.pdf
Personnel 12

GMP/EU Annex 1-2020 Draft

4.7 There should be systems in place for disqualification of personnel from entry into cleanrooms based
on aspects including ongoing assessment and/or identification of an adverse trend from the personnel
monitoring program and/or after participation in a failed APS. Once disqualified, retraining and
requalification should be completed before permitting the operator to have any further involvement in
aseptic practices.
4:15 Clean area clothing should be cleaned in a dedicated laundry facility using a qualified process
ensuring that the clothing is not damaged and/or contaminated by fibres and particles during the laundry
process. Inappropriate handling and use of clothing will damage fibres and may increase the risk of
shedding of particles. After washing and before packing, garments should be visually inspected for
damage. The garment management processes should be evaluated and determined as part of the
garment qualification program.
Operators performing aseptic operations should adhere to aseptic technique at all times to prevent
changes in air currents that introduce air of lower quality into the critical zone. Movement adjacent to
the critical zone should be restricted and the obstruction of the path of the unidirectional (first air)
airflow should be avoided. Airflow visualisation studies should be considered as part of the operator’s
training programme.
 How gowns affect EM results 13

• Gowns not correctly sized

• Compromised gowns, such as fraying cuffs, open seams, etc.
• No control on quality
• No control over the number of laundries including detergents
• Inappropriate storage of gowns
• Inadequate gowning procedures
Premises 14

GMP/EU Annex 1-2020 Draft

5.1 The manufacture of sterile products should be carried out in appropriate

cleanrooms, entry to which should be through changing rooms that act as airlocks
for personnel and airlocks for equipment and materials. Cleanrooms should be
maintained to an appropriate cleanliness standard and supplied with air which has
passed through filters of an appropriate efficiency.

Controls and monitoring should be scientifically justified and capable of evaluating

the state of environmental conditions for cleanrooms, airlocks and pass-throughs
used for material and equipment transfer.
Premises 15

GMP/EU Annex 1-2020 Draft

5.12 Airflow patterns within cleanrooms and zones should be visualised to

demonstrate that there is no ingress from lower grade to higher grade areas and
that air does not travel from less clean areas (such as the floor) or over operators
or equipment that may transfer contaminant to the higher-grade areas. Where
air movement is shown to be a risk to the clean area or critical zone, corrective
actions, such as design improvement, should be implemented.

Airflow pattern studies should be performed both at rest and in operation (e.g.
simulating operator interventions). Video recordings of the airflow patterns
should be retained. The outcome of the air visualisation studies should be
considered when establishing the facility's environmental monitoring program.
Premises 16

GMP/EU Annex 1-2020 Draft

5.4 Grade B area: For aseptic preparation and filling, this is the background
cleanroom for the Grade A zone (where it is not an isolator). When transfer holes are
used to transfer filled, closed products to an adjacent cleanrooms of a lower grade,
airflow visualization studies should demonstrate that air does not ingress from the
lower grade cleanrooms to the Grade B.

Pressure differentials should be continuously monitored. Cleanrooms of lower grade

than Grade B can be considered where isolator technology is used.
Poor RABS Design 17

Aseptic Filling Operation:

“Your firm failed to use equipment in the manufacture, processing, packing, or
holding of drug products that is of appropriate design, adequate size, and suitably
located to facilitate operations for its intended use and for its cleaning and
maintenance. (21 CFR 211.63)
The sterile filling line for injectable products lacks unidirectional airflow in the ISO
5 aseptic filling zone. The RABS airflow in the filling zone is not sufficiently robust
to protect the sterile injectable product during interventions involving operator
entry into the aseptic filling line.
Smoke studies demonstrated that the filling line design permits turbulence above
and below open vials. Opening the enclosure significantly disrupts airflow. This
turbulent air in the aseptic filling zone poses a significant contamination hazard.
Uncleanable Surfaces 18
 Common Unrecognized Issues 19

“It is useful to assume that the

operator is always contaminated
while operating in the aseptic area.

If the procedures are viewed from

this perspective, those practices
which are exposing the product to
contamination are more easily
identified.”

Hank Avallone – 1989

“a gowned operator may release as

many as 10,000 CFU/hour or
more…

(Reinmuller and Ljungqvist).

AFV/Smoke Studies is the MOST Misunderstood CR Test! 20

Smoke Studies are often approached as a “Rubber Stamp Test”, often conducted under the
assumption that they will pass because other cleanroom tests have passed.
• Air Volume and Air Velocity
• Particle Count
• Differential Pressure
• Filter Integrity
This Approach coupled with other factors contribute to Smoke/AFV Study Errors that lead
to:
• Contamination of Sterile Products
• Inspectors Comments
• Warning Letters
Frequent Inspector’s Comments on Smoke Studies 21

• Smoke Studies not conducted during dynamic conditions simulating operations with equipment
running, filling and stoppering
• Only Grade A areas and not support Grade B areas tested
• Only one angle recorded during dynamic smoke studies
• Smoke Study Video not available for inspectors
• Dynamic Smoke Study Video does not reflect actual operations as indicated in Media Fill
• Dynamic Smoke Study does not simulate all interventions
• Smoke Generator does not generate adequate smoke to evaluate the aseptic process
• Smoke Manifold is not over the Operators during the intervention
• Smoke Studies do not fully demonstrate air flow movement away from work surfaces during
interventions
• Turbulence observed with no corrective action identified
FDA 483 Observation 22

FDA 483 Observation: “Equipment for adequate control over air pressure is not provided
when appropriate for the manufacture, processing, packing or holding of a drug product.”
• Specifically your firm lacks a system of continuous monitoring of differential pressure limits
during aseptic processing of drugs intended to be sterile.
• Your current practice is to log the differential pressure readings from your positive pressure
differential pressure gauges, representing the differential pressure across your clean room
and adjacent gowning room.
• Because these gauges are located outside of the clean room itself, if a loss of positive
pressure in the clean room occurred during aseptic processing, you may not notice until the
clean room differential pressure gauges are read again.
• Your current clean room differential pressure system has no audible alarm; thus, transient
excursions of ~pressure would not be observed or recorded.
 Poor Cleanroom/Barrier23

Integration Limits the HEPA

Filtered Air Delivered into the
Grade B Support Area

Grade A Grade B
Air Inlet to Air Supply
LAF
 24

Grade B HEPA Grade B HEPA

Supply Air Supply Air

Smoke Open Active RABS RABS

KEY: UDAF
Manifold Air Inlet
First Air Position for
documenting
Non-First “FIRST AIR”
Air Inside RABS
First Air is Flowing
Air That Should Over
be Considered Product Contact
Contaminated
Surfaces
When the RABS
Door is Closed

Door Closed
Condition

Air Air
Return Return
 25

Grade B HEPA Grade B HEPA

Supply Air Supply Air

RABS ”SHORT CIRCUIT’

KEY: Open Active RABS

UDAF Air Inlet HEPA Filtered Air
intended for keeping
Smoke
First Air Manifold
Grade B area clean is
Position for re-directed to the
Inside RABS RABS inlet. Reducing
Non-First the contamination
Air Inside RABS control for the Grade
B Zone.

Air That Should

be Considered Air That Should Be
Contaminated Considered
Contaminated is
Flowing from the
floor and across the
Operator.

The Inlet of the RABS

is stronger than the
HVAC Air Return.
Creating an upwards
air flow pattern that
is DETRIMENTAL to
Poor contamination
Cleanroom/Barrier control
System Integration

Air Air
Return Return
Biological Safety Cabinet 26
 Case Study: Poorly Designed Sterile Filling Line 27

Cleanroom was Qualified including Air Flow

Visualization Studies.
• EM Data from Media Fills (Settle Plates)
were registering CFUs.
• The sample locations were suspect;
however the picture tells us everything.
• NO separative airflow between the RABS
and the operating personnel.

27
 Barrier System Flaw: Poor Cleanroom / Open
28
Passive RABS Integration

Passive RABS recycles air from the surrounding environment.

RABS Air Inlet Creates Upwards
Air Flow Open Passive RABS via LAF:
Only the LAF for the Filling
Machine Nothing for the Room LAF Unit
HEPA HEPA
Membrane diffuser
7.6 -15 cm gap between HEPA
Filter and top of RABS
No Pressure Differential
Between RABS and Where
Operators are Working GAP Between Filters
Causes Turbulance

Low Wall Returns Low Wall Returns

 ISO Doctrine of Contamination Control 29

ISO 14644-7 Requalification

(Periodic Testing or After Corrective Actions)
Seperative devices,
ISO 14644-1
(clean air hoods, Classification of
gloveboxes isolators Air Cleanliness
and mini- by
Specifications for Particle
environments) Cleanroom Class Concentration

Cleanroom
ISO 14644-4 Monitoring
Qualification
Cleanroom Risks
Testing ISO 14644-5 ISO 14644-2
Design,
Construction Cleanroom Monitoring &
Pre Tests
& Startup Operations Periodic Testing
Continuous Operation
IF Routine or
Continuous
Monitoring Data is
within Specification
Specification for ISO 14644-3
Additional Tests Test Methods:
(Particle, Temp
RH, Δ Pressure)
...
Monitoring Additional Cleanroom Parameters
& Qualification

Specification for Monitoring & Periodic Testing

Particle Monitoring 30

Environmental Monitoring as Addressed in Draft Annex 1

6.9 Particle counters should be qualified (including sampling tubing).
Portable particle counters with a short length of sample tubing should
be used for qualification purposes. Study performed for
evaluation resulted in:
• Company was using 3
types of particle
monitoring devices
• 50% discrepancy in
particle transport
efficiencies
• Discrepancy in
reading from system
to system
Particle Monitoring 31

Regulatory Thinking on Particle Sample Tubing:

GMP Annex 1: Clause 11
Cleanroom and clean air device monitoring
• “The system selected must be appropriate for the particle size considered.
• Where remote sampling systems are used, the length of tubing and the radii of any bends in the
tubing must be considered in the context of particle losses in the tubing.“
PIC/S Recommendation: “GMP Annex 1 Revision 2008, Interpretation of Most Important Changes for
the Manufacture of Sterile Medicinal Products” January 2010
• “Recommendation: This section addresses concerns especially for the sedimentation of 5 µm particles
in remote systems (as a rough example, s-shaped bent tubing of 1.5 m length can already absorb
about 30% of the 5 µm particles.)”
• “The company must qualify their particle sampler and sampling system for both particle sizes, 0.5 µm
and 5 µm.”
Particle Monitoring 32

Sample Tubing for NV Particle Counting:

ISO 14644-1:2015 for Particles >5 micron, no greater than 1 meter
1 Bend Radius (>15cm)
1 ASTM F50-12:2015 Standard Practice for Continuous Sizing and Counting of
Airborne Particles in Dust-Controlled Areas and Clean Rooms Using
Instruments Capable of Detecting Single Sub-Micrometer and Larger Particles
483 regarding non-viable monitoring 33

“No representative non-viable particle (NVP) monitoring data supports your current ISO-
5 classification for the product path from the (b)(4) to the (b)(4), which transfers product
to the (b)(4) during aseptic processing of finished drug products.
During our inspection, we documented that your NVP probes are placed (b)(4) surface
instead of near the working area. Placing the probe (b)(4) instead of near the working
area means you are unable to detect NVPs where sterile drugs are exposed during
aseptic processing.
Additionally, transferring (b)(4) vials from the filling suite to the (b)(4) can take up to
(b)(4). This extended exposure time may increase contamination hazards. However, your
firm lacks adequate environmental monitoring of this part of the operation. It is essential
that your sampling plan include areas where (b)(4) and product are exposed to the
environment, and at greater risk of contamination.”
Particle Size Capture 34

• Function of the nozzle size and air flow through the impactor, as this determines
velocity through a given nozzle (Slit or Hole).
• Distance between nozzle exit and collection surface.
• Smaller Holes and Higher Flow Rates Collect Smaller Particles.
Viable and non-viable environmental monitoring 35

GMP/EU Annex 1-2020 Draft

9.4 Risk assessments should be performed in order to establish a comprehensive

environmental monitoring program, i.e. sampling locations, frequency of
monitoring, monitoring method used and incubation conditions (e.g. time,
temperature(s), aerobic and/or anaerobic conditions). These risk assessments
should be conducted based on detailed knowledge of; the process inputs and final
product, the facility, equipment, specific processes, the operations involved,
historical monitoring data, monitoring data obtained during qualification and
knowledge of typical microbial flora isolated from the environment. Consideration
of other information such as air visualization studies should also be included. These
risk assessments should be reviewed regularly in order to confirm the effectiveness
of the site’s environmental monitoring program. The monitoring program should be
considered in the overall context of the trend analysis and the CCS for the site.
Viable and non-viable environmental monitoring 36

GMP/EU Annex 1-2020 Draft

9.7 The monitoring of Grade A zones should demonstrate the maintenance of

aseptic processing conditions during critical operations. Monitoring should be
performed at locations posing the highest risk of contamination to the sterile
equipment surfaces, container, closures and product.

The selection of monitoring locations and the orientation and positioning of

sampling devices should be justified and appropriate to obtain reliable data from
the critical zones.
Viable and non-viable environmental monitoring 37

GMP/EU Annex 1-2020 Draft

Note 1: The particulate limits given in the table for the “at rest” state should be achieved after a short “clean up” period
(defined during qualification with a guidance value of 15 to 20 minutes) in an unmanned state, after the completion of
operations (refer to paragraph 4.30 and 4.31).

Note 2: With regards to the monitoring of airborne particulates ≥5 μm particulate concentration, the limit of 29 (Grade A) is
selected due to the limitations of monitoring equipment. Alert levels should be set based on historical data, such that
frequent sustained counts below the action limit which may be indicative of system contamination or deterioration
should trigger an investigation. For the Grade A zone and Grade B area the importance of monitoring the ≥5 μm
particulates is to identify negative trends as defined in the manufacturer's CCS.
Premises: Cleanroom & Clean air device Qualification 38

GMP/EU Annex 1-2020 Draft

5.25 For cleanroom classification, the airborne particulates equal to or greater than 0.5 and
5 µ m should be measured. For Grade A zone and Grade B at rest, classification should
include measurement of particles equal to or greater than 0.5 µ m; however, measurement
using a second, larger particle size, e.g. 1 µ m in accordance with ISO 14644 may be
considered. This measurement should be performed both at rest and in operation. The
maximum permitted airborne particulate concentration for each grade is given in Table 1.
How can you monitor what you haven’t qualified?

38
Viable and non-viable environmental monitoring 39

GMP/EU Annex 1-2020 Draft

9.15 The Grade A zone should be monitored continuously (for particulates ≥0.5 and ≥5 µ m)
and with a suitable sample flow rate (at least 28 litres (1ft3) per minute) so that all
interventions, transient events and any system deterioration is captured. The system
should frequently correlate each individual sample result with the limits in Table 6 at
such a frequency that any potential excursion can be identified and responded to in a
timely manner. Alarms should be triggered if alert levels are exceeded. Procedures
should define the actions to be taken in response to alarms including the consideration
of additional microbial monitoring.
9.16 It is recommended that a similar system be used for Grade B area although the sample
frequency may be decreased. The Grade B zone should be monitored at such a
frequency and with suitable
sample size that the programme captures any increase in levels of contamination and
system deterioration. If alert or action levels are exceeded, alarms should be triggered.
Viable and non-viable environmental monitoring 40

GMP/EU Annex 1-2020 Draft

9.18 The selection of the monitoring system should take into account any risk presented by
the materials used in the manufacturing operation (for example, those involving live
organisms, powdery products or radiopharmaceuticals) that may give rise to biological or
chemical hazards.
9.21 The size of monitoring samples taken using automated systems will usually be a
function of the sampling rate of the system used. It is not necessary for the sample volume
to be the same as that used for formal classification of cleanrooms and clean air equipment.
Monitoring sample volumes should be justified.
9.24 Monitoring conditions such as frequency, sampling volume or duration, alert levels and
action limits and corrective actions (including an investigation) should be established
in each manufacturing area based on data generated during the initial qualification process,
ongoing routine monitoring and periodic review of data.
Viable and non-viable environmental monitoring 41

GMP/EU Annex 1-2020 Draft

9.25 Where aseptic operations are performed, microbial monitoring should be frequent using a
combination of methods such as settle plates, volumetric air sampling, glove, gown and surface
sampling (e.g. swabs and contact plates). The method of sampling used should be justified within
the CCS and should be demonstrated not to have a detrimental impact on Grade A and B airflow
patterns.
9.27 Continuous viable air monitoring in the Grade A zone (e.g. air sampling or settle plates)
should be undertaken for the full duration of critical processing, including equipment (aseptic set-
up) assembly and filling operations. A similar approach should be considered for Grade B
cleanrooms based on the risk of impact on the aseptic processing. The monitoring should be
performed in such a way that all interventions, transient events and any system
deterioration would be captured and any risk caused by interventions of the monitoring
operations is avoided.
9.29 Sampling methods and equipment used should be fully understood and procedures should
be in place for the correct operation and interpretation of results obtained. The recovery
efficiency of the sampling methods chosen should be qualified.
Viable and non-viable environmental monitoring 42

9.31 Action limits for viable particle contamination

(a) Settle plates should be exposed for the duration of operations and changed as
required after 4 hours (exposure time should be based on validation including
recovery studies and it should not have any negative effect on the suitability of the
media used). Individual settle plates may be exposed for less than 4 hours.

(b) It should be noted that for Grade A, any growth should result in an investigation.
Viable and non-viable environmental monitoring 43

9.33 Microorganisms detected in Grade A zone and Grade B area should

be identified to species level and the potential impact of such
microorganisms on product quality (for each batch implicated) and
overall state of control should be evaluated.

Consideration should also be given to the identification of

microorganisms detected in Grade C and D areas (for example where
action limits or alert levels are exceeded or where atypical or potentially
objectionable microorganisms are recovered). The approach to organism
identification and investigation should be documented.
Quality Control 44

10.9 Media used for environmental monitoring and APS should be tested for its
growth promotion capability, in accordance with a formal written program

10.10 Environmental monitoring data and trend data generated for classified areas
should be reviewed as part of product batch certification. A written plan should be
available that describes the actions to be taken when data from environmental
monitoring are found out of trend or exceeding the established limits. For
products with short shelf life, the environmental data for the time of
manufacture may not be available; in these cases, the certification should include a
review of the most recent available data.

.
Quality Control of Media 45

Accuracy of viable monitoring results depend upon media quality and incubation. Common
causes for erroneous results :
• Discoloration or hemolysis
• Storage location
• Integrity of packaging
• Broken or cracked petri dishes
• Quality and accuracy of labeling
• Condensation in petri dishes
• Retracted medium
• Dried and cracked media
• Sloped or uneven filling of petri dishes
• Contamination
• Gel strength
• Pitted surface or large bubbles
• Presence of leakage….
Risk Based EM Program 46

Data should be reviewed continually, not only during batch release. The purpose of EM is to ensure
controls are working and there is no contamination risk to product.

Microbial identification should be considered as a critical component of a risk-based monitoring program.

Though each colony from plates may not be identified; only unique colonies may be identified, however
the counts of each unique type of colony should be recorded. This should be considered during trending
to understand contamination sources and remedial actions necessary.

Trends; beyond the data for each room, should depict excursions, investigations, remedial actions, etc.

Predominance of microorganisms in each room should be assessed to evaluate risk to product and
patient.

This information should be used to improve cleaning procedures, evaluate facility or operational flaws.

Compromised agar media should not be used for monitoring; this requires a comprehensive quality
control program for incoming media.
FDA 483 Observation 47

Your firm failed to establish laboratory controls that include scientifically sound
and appropriate specifications, standards, sampling plans, and test procedures
designed to assure that components, drug product containers, closures, in-
process materials, labeling, and drug products conform to appropriate
standards of identity, strength, quality, and purity (21 CFR 211.160(b)
For example, during inspection of the QC microbiology testing laboratory, our
investigators observed:

A. No growth on the positive control plate for media used to test

microbiological (b)(4) samples. When a positive control fails to yield growth,
test results cannot be considered valid due to the potential for false negatives.
FDA 483 Observation 48

B. Desiccation of a contact media plate used during environmental monitoring

of the sterility testing area. Desiccated, cracked, or otherwise damaged (b)(4)
compromises microbial growth promotion and accurate enumeration, and can
lead to artificially low microbiological counts and false negatives. Using
deficient media compromises the validity of your microbiological test results.

Also, you did not appear to routinely identify (i.e., to species level) bacterial
and fungal isolates recovered during environmental monitoring of your aseptic
processing room.

C. Air bubbles between filtration (b)(4) and (b)(4) plates in 13 out of (b)(4)
microbiological (b)(4) system sampling plates. Inadequate contact between the
filter (b)(4) and the (b)(4) plate may compromise recovery.
Fungal ID System and Methodology 49

• Capable of
identifying
Presence maximum Polyphasic
or absence organisms Approach
Images
• Can identify
consistently Keeping up
ID Library/ Reliability
with
System Database
Taxonomic
Taxonomy is important • How was the Changes
If you don’t have consistent names, library created
trending or investigations have no Diversity
value!!

Multiple ID systems is not the solution-

final product and EM IDs should be
correctly and consistently identified!!

49
Reasons for Environmental Monitoring 50

• Evaluate Environmental Controls

• Evaluate Facility Condition
• Evaluate Personnel Behavior
• Evaluate Gowning Procedures
• Evaluate Aseptic Techniques
• Evaluate Risk to Product
• Patient Safety
 51

Questions??