Microbiological

Status of Lab Animals

Including
Gnotobiology
Prof. Dr. Khaled Al-Amry
 Isolators Attributes

It may be a small or large container, usually made of

glass and steel or plastic, with plastic or rubber
gloves.
 Isolators Attributes

Food and utensils are sterilized and taken into the

isolator through an adjoining area called the sterile
lock.
 Air handling system
The isolator has an air handling system
Positive/Negative-pressure isolator
Sterilization of isolators
Metal isolator
Plastic flexible pharmaceutical isolator
Sterilization
Sterility testing
 Isolators Attributes

Since the interior of the isolator is sealed off from the

background cleanroom, operator access to the interior
is done through glove ports or half suits.

Sterile containers, stopper components, and

environmental monitoring materials are brought in to and
out of the isolator through air locks, mouse holes, and
devices known as Rapid Transfer Ports (RTPs)
 Isolators Attributes
One of the major advantages that isolators have over RABS type systems is
Microbiological status of the isolator
Isolator
Aseptic filing
The “interface”
Interface isolator
Plastic glove isolator
Integrity checks
The “interface”
Other parameters which could influence and impact the
decontamination effect
Controlling the microbiological status of an operating isolator
 Air & Surface monitoring
Air monitoring
• is performed with the semi-
quantitative settling plate
method (which are exposed to
the environment for a defined
(and validated) time span) or
the quantitative air sampling
method, e.g. by the impaction
method.
• on a regular basis, an
anaerobic microbiological
monitoring should be
performed to produce data on
the anaerobic status of the
filling line.
Surface monitoring
• due to the nature of the methods
used for microbiological
monitoring, a continuous control is
not possible for surface monitoring
•
contact plates for flat surfaces or
the swab technique for irregular
surfaces are used
• a monitoring of surfaces during
production is not recommended, as
additional interventions during the
filling are needed for sampling and
residuals of nutrient medium can
endanger the asepsis of the interior
of the isolator.
Environmental monitoring
Sampling points
Sampling points
 Isolator
Aseptic isolator with a, six-axis robot arm that is compatible with vapor phase hydrogen
peroxide sterilization
Restricted Access Barrier Systems (RABS) Attributes

Unlike isolators they use a combination of

physical and aerodynamic barriers to prevent ingress of
contaminants into the interior environment.

The physical barrier is similar to machine guarding having glass or

polycarbonate doors with stainless steel walls that totally enclose the
machinery with an air handler supplying HEPA filtered, unidirectional
airflow providing an ISO 5 environment.
Restricted Access Barrier Systems (RABS) Attributes

RABS are typically unsealed barriers having the HEPA filtered air
supplied to the RABS interior and exhausted through a gap
between the RABS walls and the equipment.

RABS that exhaust to the background environment are

referred to as open RABS.
Restricted Access Barrier Systems (RABS) Attributes

Passive open RABS is a barrier system that is built around

equipment installed below air handlers in the background
cleanroom, which provide the ISO 5
environment.

Closed RABS offer another option and are by design sealed

isolators that can be positive or negative pressure, but are
manually cleaned and bio-decontaminated rather than utilizing an
automated bio-decontamination process typical of isolators.
Restricted Access Barrier Systems (RABS) Attributes

Glove ports and half suits are also used to further

separate an operator from the sterile interior of the
RABS.

In the early implementation of the RABS concept by the

pharmaceutical industry, many companies considered any enclosure
around the process a RABS, claiming that it provided enhanced
sterility protection over conventional clean room manufacturing.
 Restricted Access Barrier Systems (RABS) Attributes

But in many cases operators would continually open the

RABS doors to access areas within the critical zone to
perform an intervention. Thus defeating the purpose of the
RABS and creating an unpredictable level of contamination

Ultimately, RABS can meet the

requirements of Advanced Aseptic Processing if it is
used as intended; separate the
operator from the process.

RABS can only achieve this criterion when any and all
open door interventions are prohibited.
Restricted Access Barrier System
Restricted Access Barrier System (RABS) encloses a vial filling, and helps
to ensure the aseptic environment
Robotics in Aseptic Processing
 Robotics in Aseptic Processing
robot safety compliant RABS unit that protects the operator from the robot system and protects the product from contamination originating from the operator
The combination of Isolator-Barrier technology with robot
system
 Microbiology Lab Practices and
Safety Rules

Wash your hands when you arrive at the lab and

with disinfectant soap again before you leave.

Do not put anything in your

Absolutely no food, drinks,
chewing gum, or smoking
is allowed in the laboratory
mouth such as pencils, pens,
labels, or fingers. Do not store
food in areas where
microorganisms are stored.
 Microbiology Lab Practices and
Safety Rules

This garment must: cover your

Purchase a lab coat and arms, with long sleeves, with
safety glasses, bring buttons or snaps and be able to
them to class, and use
them
be removed without pulling it
over your head. Leave
protective clothing in the lab
and do not wear it to other non-
lab areas.

Avoid loose fitting

Wear appropriate shoes
items of clothing
(sandals are not allowed) in the
laboratory.
 Microbiology Lab Practices and
Safety Rules

Backpacks, purses, and coats

should be placed in the
Keep your workspace
free of all unnecessary cubbyholes by the front door
materials of the lab. Place needed items
on the floor near your feet, but
not in the aisle.

Laboratory equipment and

work surfaces should be
Disinfect work areas decontaminated with an
before and after use appropriate disinfectant on a
with 70% ethanol or
fresh 10% bleach routine basis, and especially
after spills, splashes, or other
contamination.
 Microbiology Lab Practices and
Safety Rules

This saves a lot of time and

Label everything
clearly
effort, and keeps you away
.from many accidents

Replace caps on
reagents, solution
Do not open Petri dishes in the
bottles, and bacterial lab unless absolutely
cultures necessary.
 Microbiology Lab Practices and
Safety Rules

You can simply impress your

Sterilize Inoculating
loops and needles in a audience and add a unique
Bunsen burner before zing and appeal to your
you lay them down
Presentations.

when not is Turn off Do not leave open flames

Bunsen burners use unattended .
 Microbiology Lab Practices and
Safety Rules

Be sure to remove all papers,

When you flame sterilize
with alcohol, be sure notebooks, combustible
that you do not have any materials and excess
.papers under you
chemicals from the work area.

Treat all Use appropriate care and do

microorganisms as not take cultures out of the
potential laboratory.
pathogens
 Microbiology Lab Practices and
Safety Rules

If you are working with a

Wear disposable sample that may contain a
gloves when working
with potentially
pathogen, then be extremely
infectious microbes or careful to use good
samples
bacteriological technique.

Sterilize equipment and Long hair must be restrained if

materials Bunsen burners are in use.
 Microbiology Lab Practices and
Safety Rules

Mouth- pipetting has been

known to result in many
Never pipette by laboratory-acquired infections.
mouth. Use a
pipetting aid or
With the availability of
adjustable volume mechanical pipetting devices,
pipettors mouth pipetting is strictly
prohibited.

Do not pour anything down the

Consider everything a sink. Autoclave liquids and
biohazard broth cultures to sterilize them
before discarding.
 Microbiology Lab Practices and
Safety Rules

Dispose of all solid

And autoclave it before
waste material in a
biohazard bag discarding in the regular trash.

Familiarize yourself (e.g., eye-wash station,

with the location of shower, sinks, fire extinguisher,
safety equipment in
the lab biological safety cabinet, first
aid kit, emergency gas valve).
 Microbiology Lab Practices and
Safety Rules

As with unconventional glass,

Dispose of broken wrap the fragments in paper
glass in the broken and seal them tight if you use
glass container plastic bags.

Dispose of razor blades, protect waste handlers from

syringe needles, and sharp both physical and
metal objects in the contamination hazards that
“sharps” container they present.
 Microbiology Lab Practices and
Safety Rules

Report spills and Clean small spills with care

accidents immediately to (see instructions below). Seek
your instructor help for large spills.

Report all injuries or

No matter how small they
accidents immediately to
the instructor seem.
Laboratory safety equipment
Safety shower

Safety can
Eyewash station
Causes of Contamination of Laboratory Samples and their Prevention
Causes of Contamination of Laboratory Samples and their Prevention
Causes of Contamination of Laboratory Samples and their Prevention
Causes of Contamination of Laboratory Samples and their Prevention
Causes of Contamination of Laboratory Samples and their Prevention
Causes of Contamination of Laboratory Samples and their Prevention
Causes of Contamination of Laboratory Samples and their Prevention
Thank you