WORLD

HEALTH

ORGANISATION

DISTR. : LIMITED
DISTR. : LIMITEE

ORGANIZATION
MONDIALE

DE LA

SANTE
WHO/Rabies/90.201
ORIGINAL:

Rev.1
FRENCH

INSTRUCTIONS FOR THE DESIGN, EQUIPPING AND STAFFING

OF A LABORATORY FOR THE PRODUCTION OF ANIMAL RABIES VACCINE
PREPARED ON CELL-CULTURE
by Dr P. Reculard, Short term consultant to
Veterinary Public Health Unit, WHO

This document
is not issued to the general public, and
all rights are reserved by the World Health Organization
(WHO).
The document may not be reviewed, abstracted,
in part or in whole,
quoted, reproduced or translated,
No part
without
the prior written permission of WHO.
of this document may be stored in a retrieval system or
transmitted
in any form or by any means - electronic,
mechanical or other - without the prior written permission
of WHO.

Ce document nest pas destine a ltre distribue au grand public

et tous les droits y afferents sont reserves par IOrganisation
mondiale de la Sante (OMS). II ne peut etre comment&
resume, cite, reproduit ou traduit, partiellement ou en totalite,
saris une autorisation prealable &rite de IOMS. Aucune partie
ne doit btre chargee dans un systeme de recherche documentaire ou diffusde sous quelque forme ou par quelque moyen
que ce soit
Clectronique, mecanique, ou autre - saris une
autorisation prealable &rite de IOMS.

The views expressed in documents by named authors are solely

the responsibility of those authors.

Les opinions exprimees dans les documents par des auteurs

cites nommement nengagent que lesdits auteurs.

WHO/Rabies/90.201

Rev.1

This
document
describes
the
requirements
for establishing
a unit
capable
of an annual production
of about half
a million
doses of cell-cultured
rabies
vaccine
for veterinary
use.
It is intended
as a guide
for decision
makers wishing
to study the feasibility
of
and
at
the
same
time
provides
such
a project,
instructions
for
laboratory
personnel
planning
to
improve
or establish
such a unit.
The use of roller
bottles
for antigen
production
is
replacement
by classical
fermentor
described.
However,
technology
is recommended
if a higher
annual
output
is
required.
This techonology,
however,
calls
for a high
level
of investment.
New technologies
are currently
being tested
and may
become available
in the not too distant
future.
These
technologies
may be less
demanding
in terms
of space
and laboratory
environment
control.
Nevertheless,
the
cycles
of manufacture
and control
remain
more or less
whichever
procedure
is used.,
Therefore,
the
same,
should
be
possible
without
major
conversion
any
modification
to either
the design
or the specifications
of the laboratories
described
in this
document.

This

document was prepared

under the direction
of
Dr F.-X.
Meslin,
Veterinary
Public
Health,
Division
of Communicable
Diseases,
WHO, Geneva,
under the auspices
of the
Mediterranean
Zoonoses
Control
Programme

WHO/Rabies/90.201

Rev.1
page 1

INSTRUCTIONS FOR THE DESIGN, EQUIPPING AND STAFFING OF FACILITIES

FOR PRODUCTIONOF RABIES VACCINE FOR VETERINARY USE FROM CELL CULTURES

LIST OF CONTENTS
Page
. ..................................

FOREWORD...................................
OBJECTIVES ....................................................................
Chapter

1:

1.1
1.2
1.3
1.4
1.5
Chapter

2:

vaccine

......................................

Control

of rabies

vaccine

.........................................

Introduction
........................................................
Buildings
...........................................................
Equipment ...........................................................
Staff
...............................................................
3:

3.1
3.2
3.3
3.4
3.5
Chapter

of rabies

Introduction
........................................................
Production
site .....................................................
Buildings
...........................................................
Equipment ...........................................................
Staff
...............................................................

2.1
2.2
2.3
2.4
Chapter

Production

Central

Services

..................................................

Introduction
........................................................
Central Washing Plant ...............................................
Media Preparation
Unit ..............................................
Filling
and Packaging Unit ..........................................
Other General Services
..............................................
4:

Animal

houses

.....................................................

2
3
4
4
4
5
9
10
13
13
13
14
15
16
16
16
17
19
21
24

........................................................
Introduction
Central animal houses for small laboratory
animals ..................
Animal houses for testing
of rabies vaccine for veterinary
use ......

24
24
24

Annex 1

List

used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

28

Annex 2

Plans and figures

...................................................

39

4.1
4.2
4.3

of equipment

and materials

*********

WHO/Rabies/90.201
page 2

Rev.1

FOREWORD
The development of cell culture
techniques
and progress
the last few decades have led to the development of new viral
antigens
(complete virions
or subunits)
harvested
in culture
cells with the chosen virus.
Human and animal rabies
technological
progress.

vaccines

are among those

that

made in virology
over
vaccines that use
after the infection
of
have benefited

from this

most of the rabies vaccines used throughout

However, in spite of such progress,
the world are still
prepared from nerve tissue taken from experimentally
infected
They consist
of a suspension of cerebral
matter obtained from homogenized
animals.
Large quantities
of vaccine can be produced
brains of sheep, lambs and newborn mice.
in a conventional
laboratory
with simple, rudimentary
of equipment, which means they
are produced cheaply.
On the other hand, and whatever the scale of production,
manufacture
of rabies
vaccine from cell cultures
requires
purpose-built,
protected
premises for each of a
It calls for special
equipment and infrastructure,
number of different
functions.
and staff
qualified
and experienced
in cell and virus culture
considerable
resources,
techniques.
the World Health Organization
(WHO) recommends that
Nevertheless,
rabies vaccines prepared from cell cultures
be used wherever possible,
These are:
advantages they offer.
1.

Safety

inactivated
because of the

and innocuitv

The absence of nerve tissue considerably

reduces the risks of allergic
reactions
Furthermore,
and serious neurological
damage, which is most often caused by myelin.
it is easy to check the complete inactivation
of virus from cell cultures,
to ensure
freedom from residual
pathogens.
2.

Potencv

Successive improvements in manufacturing

procedures now yield inactivated,
stabilized
and adjuvanted
rabies vaccines from cell cultures
with potency higher than
Animals can be immunized for at least two years with
that of brain tissue vaccines.
a single dose of these new vaccines.
Special attention
is therefore
given in these Instructions
to the resources
required
for the production
of rabies vaccines from animal cells cultivated
and
infected
in vitro.
the Instructions
concern only the requisite
conditions
for
Furthermore,
adjuvanted
rabies vaccine for the protection
of animals.
preparation
of liquid,
Tl=Y
therefore
do not deal with the installations
and equipment used for purifying
or
freeze-drying
the virus,
features
which are indispensable
to the production
of rabies
vaccine for human use.

WHO/Rabies/90.201

Rev.1
page 3

OBJECTIVES
These Instructions

aim to help Member States

of unit

they need for

to:

(i)

decide what size

veterinary
use;

production

of rabies

vaccine

for

(ii)

devise the production

unit as part of a larger facility
that should have
sufficient
infrastructure
and resources to guarantee the quality
of the work
done there;

(iii)

choose the form and layout of buildings,

and also the equipment, materials
and staff
that provide the best in terms of safety and effectiveness.

Repeated reference
is made in these Instructions
to the technical
recommendations
that have appeared in WHOreports,
especially
the following:
WHOTechnical
Report
Committee on Rabies);

No. 709, 1983 (Seventh

Series,

report

of the WI-IOExpert

report of the WHO

WHOTechnical
Report Series, No. 658, 1981 (Thirty-first
Expert Committee on Biological
Standardization),
Annex 3 - Requirements for rabies
vaccine for veterinary
use.
Those reports

should

be consulted

in conjunction

with

these

instructions.

the mention of manufacturers

or suppliers
in these Instructions
Finally,
pages 28-38) should not be taken as an
list
of equipment and main supplies,
expression
of preference
by WHO: they are cited only by way of example.
These Instructions

comprise

Chapter

1:

Production

Chapter

2:

Control

of rabies

Chapter

3:

Central

services

four

of rabies

parts:

vaccine

vaccine
(in

common with

other

production

units):

central
washing plant
media preparation
unit
bottling
and packaging unit
central
warehouse
administration
essential/common
services
Chapter

4:
central
animal

Animal

houses

animal houses
houses for testing

of rabies

vaccine

for

safety

and efficacy.

(see

WHO/Rabies/90.201
-page 4

Rev.1
CHAPTER 1.

1.1

PRODUCTIONOF RABIES VACCINE

INTRODUCTION

1.1.1
The general provisions
set out in the report of the WHO Expert Committee on
Biological
Standardization
(WHO Technical
Report Series, No. 760, 1987, page 171,
paragraph 2) for manufacturing
establishments
and control
laboratories,
apply also to
laboratories
for the production
of rabies vaccines for veterinary
use.
the quantity
of equipment and the principles
of
1.1.2
The size of the premises,
operation
are determined by the manufacturing
capacity
selected,
which in our example
is between 400 000 and 500 000 ready-to-use
doses of vaccine per annum.
This objective
can be achieved at. the "semi-industrial"
level,
virus culture
technology
in roller
bottles,
within
a compact space,
a mechanized production
line.

using cell and

with no need for

1.1.3
The production
of viral
vaccines from cell cultures
is based on the seed lots
systems (see WHO Technical
Report Series, No. 709, 1983, paragraph 4.3), namely seed
viruses
from a rabies virus strain
and, when cell lines are used, cell seeds.
The quality
and quantity
of rabies virus produced
depending on the cell and virus strain
pair used.

in culture

can vary

greatly

It is therefore
important
to note that in these Instructions
the data are based
on average yields
obtained using one of, the best known systems for production
of
BHK (baby hamster kidney) cell lines infected
rabies vaccine for veterinary
use:
with the Pasteur-PV strain
of rabies virus adapted to those cells.
In that system, 3 to 5 x lo6 infected
cells provide in 1 ml of harvest enough
viral
suspension to make a dose of finished
vaccine that conforms to WHO
recommendations
(potency >l.O IV) (see WHOTechnical
Report Series, No. 709, 1983,
paragraph 5.4).
The premises are therefore
designed and equipped for the annual
500 litres
of cell and virus culture
in roller
bottles,
divided
into
to be produced 10 months in the year.
25 litres,

production
20 batches

of
of

on an industrial
scale of 1 million
doses per annum or
NOTE. For production
-*
in fermentors
is best.
Such technology
involves
very
more, the procedure of culture
It also calls for maintenance engineers,
and scientists
and
heavy investments.
technicians
who are highly
qualified
and experienced
in this field.
Yet, in this
are the same and the cycle of vaccine production
is identical
so
case, the structures
that if a simplified
method of producing cultures
in fermenting
vats were devised in
the production
premises described
in these Instructions
could still
be
the future,
used, without
significant
alterations.
the facility
for production
of rabies vaccine for
1.1.4
In these Instructions,
veterinary
use is envisaged as part of a larger set-up in which other viral
and
This is because the infrastructure
and general
bacterial
vaccines are produced.
facilities
required
are such that their cost could not be reasonably borne by
production
of rabies vaccine alone.
1.2.

THE PRODUCTIONSITE

It should of course be borne in mind that before any construction

work begins,
to existing
the site must be properly
surveyed, even for a simple extension
buildings.
The survey of a site for biological
productions
must examine the
following:

WHO/Rabies/90.201

topography
access

and planned

the risk

of the soil
of flooding

supply

electricity

in the neighbourhood;

and substrate;
and other

- quality
supply

prevention

telephone,
facilities
and solid
1.3

BUILDINGS

1,.3.1

Inclusion

climatic

hazards;

and quantity;

- voltage,

access to supplies
of liquid
pure nitrogen,
etc.;
fire

activities

roads;

the nature

water

or existing

Rev.1
page 5

available
nitrogen,

power,
dry ice,

risks

of power cuts,

etc.;

oxygen,

dioxide,

gas,

carbon

facilities;

telex,

etc.;

for decontamination
waste.

in an overall

and evacuation

of liquid

and gaseous effluents,

nlan

1.3.la
In view of the risks involved
in handling rabies virus,
the premises used for
manufacture
of the vaccine must be independent from the other buildings
and reserved
exclusively
for the use of the rabies unit for as long as production
continues.
Between two runs of production,
however, if the premises are free, they may be used
(subject
to total
disinfection)
for preparation
of another inactivated
viral
vaccine.
The premises should then be duly disinfected
before rabies vaccine
production
is resumed.
1.3.lb As stated in paragraph 1.1.4,
the building
should be part of a facility
where
several vaccines or other biological
products are manufactured.
In this way it
shares certain
services
with other units:
central
laundry,
building
management,
distribution,
packaging,
storage of finished
products,
general services,
and control
laboratories.
A practical
way of building
such a complex would be to put all the production
and control
buildings
inside a hangar, which would isolate
them from the outside
(see
plan for biological
production
unit,
plan no. 1, page 39).
Such a system facilitates
links between the units and allows the installations
to be maintained
without
entry
into the work areas.
The buildings
must have air conditioning.
This system is
particularly
recommended in countries
where temperatures
can be very high.
The
hangar creates a buffer
area between the buildings
and the outside environment.
1.3.2

Choice of tvne.of

1.3.2a

The buildings
purpose-built

construction

on the site
with

permanent

built
from prefabricated
platform;
or built
of assembled
either
as laboratories

can be:
materials;

panels

assembled

and perfectly

or transportable
modules
or as animal houses.

sealed

(see figure

on a concrete

1, page 40),

equipped

WI-lO/Rabies/90.201
page 6

Rev.1

1.3.2b Traditional
buildings
allow free choice of shape, size and configuration
This type of
the interests
of rational
connection
of one area to another.
construction
is most suited to the building
that houses the essential/common
in view of safety regulations
and the number of different
machines.
services,

in

For all the other buLldings,

in each country concerned,
the cost of traditional
It must always
construction
must be weighed against that of prefabricated
buildings.
be ensured that the firms contacted have the experience
and the resources
to provide
the quality
and standard of work required by building
specifications.
1.3.2~ Buildings
made from prefabricated
panels are easily
extended and modified.
Figures 2 and 3 on page 41 show a laboratory
built
of panels
Building
is quick.
with panes of glass,
1.2 m wide and 3.1 m high.
Such panels can be blank, or fitted
Interior
partitions
also are made with panels which are joined
windows or doors.
of
with brackets
and fixtures
In such a way as to seal each room. The flexibility
of corridors
and
the system allows for different
sizes of rooms, configurations
airlocks.
The examples annexed to these Instructions
show several buildings,
including
one
which measures approximately
30 x 13 m. In
for the production
of rabies vaccine,
1.2 m panels, this amounts to 2 x 25 panels long and 2 x 11 panels wide.
1.3.2d Another system for prefabricated
construction
is based on assembly and
juxtaposition
of modules that come in a variety
of sizes and can be combined to suit
Large modules can be partitioned
as described
above.
requirements.
Modules can also be purchased fully
equipped but even several modules together,
The modules marketed
will not be big enough for the production
of a rabies vaccine.
by the IFFA-CREDO Company, for example, are 13.5 m long and 3 m wide, which allows
(see figures
4
only for a row of two or three communicating
rooms, with no corridor
Such modules can nevertheless
be
on pages 42 and 43).
and 5, and specifications,
for special
functions
such as small, independent control
used in production
centres,
units or separate,
isolated
animal houses.
1.3.3

Main specifications
vaccine

of the buildine:

for

nroduction

of veterinarv

rabies

1.3.3a The size and layout are determined by the objective

of approximately
500 000 annual doses of vaccine produced from BHR cell line cultures
infected
by an
The plan presented in this document (see plan of unit for
adapted Pasteur strain.
production
of rabies vaccine for veterinary
use, plan no. 2, page 45) shows a
rectangular
building
30 m by 13m.
1.3.3b The building
and interior
installations
preventing
the escape of rabies
conditions,

must provide safe operating

virus and external
contamination.

should be at
1.3.3~ Whatever type of structure
is chosen, the height of the ceiling
Floor coverings
least 3 m to allow bulky equipment to be installed
and shifted.
Walls and
not giving rise to dust.
smooth and continuous,
should be non-slip,
finish
that is easy
ceilings
should be painted or plastered
with a smooth, waterproof
to clean and free of cracks.
Worktops (mobile tables and laboratory
benches) should be made of shock
They should be easily
to reduce the risk of breaking glass.
absorbing material,
disinfected
and resistant
to bleach and alcohol.
Periodic

disinfection

of the premises

must be possible.

WHO/Rabies/go.201

Rev.1
page 7

areas must be sealed and independent units equipped with their

1 . 3 . 3d The "sterile"
They are ventilated
with air that is sterilized
by passage
own airlock
entrances.
and introduced
through indeoendent and easily accessible
through absolute filters,
ventilation
shafts.
water should be supplied to these
Only pure, sterile
spilled
contain neither
sinks nor drains nor gully traps:
emptied into autoclavable
containers.
Lighting
should be either
sealed panes of glass.

electric

or natural,

sterile
liquid

through

areas, which must

must be sponged and

perfectly

airtight,

it is recommended that care and maintenance work on the plant within

Finally,
these areas be reduced to a minimum by having electrical
junction
boxes, stopcocks
controls
etc. housed in service areas accessible
for fluids
and gases, temperature
from outside.
1.3.4

Descrintion

of nremises

and annexes

1.3.4a
layout

The building
houses three separate areas.
of rooms in each area and how staff
should
ventilated

with

Plan no. 2 on page 45 shows the

move from one area to another:

- Area 1 is a clean
pressure;

area,

dust-free

air,

in slight

- Area 2, accessible
with air-conditioning,
kept under positive

via airlocks
Sl and S2, is a m-virulent
air filtered
through absolute filters
pressure P2 > Pl;

Pl

sterile
area,
(HEPA quality)
and

- Area 3 is a sterile
area where the rabies virus is handled before
inactivation;
it is therefore
a virulent
area, accessible
by airlocks
S3 and
It, too, has air-conditioning
that circulates
sterile
air.
The best way to
s4.
prevent the virus escaping this area is by using neeative pressure.
since such a system is very expensive,
the sterile
air can also
Alternatively,
be propelled
by using superpressure
P3 less than superpressure
P2, as long
as the ventilation
shafts for each area are independent,
and if the air
extracted
from the virulent
area is decontaminated
(e.g. by passage over
elements heating it to 180C).
1.3.4b

Area 1

This comprises an entrance hall,

changing rooms, two toilets,
an office,
a store
leading to the airlocks
for small equipment and supplies
in use, and two corridors
Sl and S2 to the non-virulent
sterile
area, S3 and S4 to the
where the staff
enter:
There are therefore
two airlock
doors for each section,
one for men
virulent
area.
and the other for women. Within these airtight
entrances
the staff undress, shower
and dress in the protective
clothing
prepared for them in sterile
packaging
(overalls,
boots, bonnets, protective
glasses,
disposable
masks and gloves)
(see
figures
6 and 7, page 44).
Area 1 also contains
an assembly and sterilization
room, since although there is
a central
washing plant for that purpose, it is verv stronnlv
recommended that
equipment be sterilized
in this area with a double-entry
autoclave
and oven opening
directly
on the protected,
sterile
areas where supplies
can be stored.
Experience
has shown that this prevents many kinds of "subtle"
contamination
that tend to plague
laboratories
which prepare cell cultures
in large quantities.
The central
washing
plant washes and rinses equipment, and the final
cycle of preparation,
assembly and
sterilization
takes place in Area 1 of the "rabies"
building.

WHO/Rabies/90.201
page 8

Rev.1

To avoid contamination
the media should also be filtered
and put in small
The central
media preparation
service weighs and mixes ingredients
and
containers.
then sends them, unsterile,
to Area 1 of the rabies building
to
prepares solutions,
be placed in containers
in equipment airlock
EAl.
A tube connects this area to
sterile
Area 2, where the media are sterilized,
filtered
and put in small
under a laminar airflow
extractor
fan.
receptacles,
1.3.4~

Area 2
This

comprises:

- A room for storage

the oven (F);

of sterile

material

coming out of the autoclave

(Al)

- A room for filtration

and distribution
of media coming through supply
2-position
extractor
fan with
Ml . This is equipped with a "Biohazard"
laminar airflow
cabinet for two workers;
vectical
- A cold
+8"C);

room for

- A room for

the storage

incubation

of filtered

of the media before

media (temperature
use (temperature

and
airlock

between +2" and

37" + 0.5"C);

equipped with Biohazard laminar airflow

cabinets
- A room for cell cultures,
room at 37" (+ 0.5"C)
four people (2.50 m), communicating with an incubation
containing
the mobile "Roller"
frames for roller
bottles
(see figure
8,
page 46);
- A room where the virus is transferred
after inactivation,
vat.
airlock
SM3, which is equipped with a decontamination
laminar airflow
cabinet for one person;
- A room for preparation
of the finished
vaccine (addition
This room has a 2-man vertical
airflow
cabinet
adjuvant).
a cold room for bulk storage of the finished
product.
1.3.4d

for

through a supply
This room has a
of stabilizer
and
and communicates with

Area 3

In this plan the virulent

area is quite separate from the others,
being linked
This corridor
leads to staff
airlocks
S3 and S4 and
to them by a special
corridor.
material
must pass for
double entry autoclave A2, through which all virulent
Sterile
equipment enters a
3 through
decontamination
before it leaves the area.
supply airlock
SM2, from area 2.
In area 3 there is a room for cell and virus cultures
that communicates with an
incubation
chamber whose temperature
can be set between 30C and 38C with an
This chamber contains
the mobile roller
frames that hold the
accuracy of + 0.5"C.
culture
bottles.
The culture
room communicates also with a cold room (temperature
between +2' and
+ 8'C) for the viral
harvest which is in the process of becoming inactive.
The
bottles
of inactivated
virus are passed through airlock
EA3 to the transfer
room
(end of inactivation)
in non-virulent
sterile
area 2.
rooms and cold rooms of the three areas can be either
NOTE. The incubation
built-infrom
the start or installed
by manufacturers
of collapsible
and convertible
making it easier to switch to the
The latter
option is more flexible,
chambers.
or to use.the building
for a different
purpose
production
of culture
with fermenters,
altogether.

WHO/Rabies/90.201

1.4.

EQUIPMENT

listed

The equipment required

below for each area.

1.4.1

Area 1

1.4.la

Room for

assembly

for

production

of inactivated,

absorbed

rabies

Rev.1
page 9

vaccine

is

and sterilization:

1 double-entry
autoclave with
a display
showing the cycles.

its own steam supply,

Accessory equipment:

a vacuum pump, a purger, and

2 trolleys
and 2 baskets;

100 x 60 x 120 cm, with

1 oven for sterilization
at 180C. Minimum dimensions:
Temperature programmable between 50" and 250C with
fan extraction
of hot air,
recording
of heating cycles;
120 x 80 cm, and 80 cm high;

2 trolleys:
1 mobile

1 peristaltic

200 x 80 cm, 90 cm high;

pump;

1 machine
1.4.lb

assembly,

bench for

for

Room for

sealing

liquid

plastic

nitrogen

packaging.
storage:

4 wide aperture,
40-litre
containers
equipped with canisters
for tubes or ampules of 2 and 5 ml (for cell seeds);
1 tank of liquid
connected to it
1.4.1~

Room for

nitrogen
of 200 litres
with protected
piping;

frozen

capacity,

small

1 machine

for

1 dry-ice

chest

1.4.2

Area 2

1.4.2a

Storage
1 "Milli

1.4.2b

crushed

capacity

for

storage

of

ice;

(1 m3).

sterile

Q" or equivalent

Room for

300 litres

equipment:

room for

2 trolleys

the room and

seeds) of approximately
350 litres
capacity,
or liquid
carbon dioxide safety device;

1 freezer
at -20C (+ 5C) of approximately
beta-propiolactone
etc.
sera, antibiotics,
Room for

outside

stocks:

2 freezers
at -85C (for viral
equipped with a liquid
nitrogen

1.4.ld

situated

(4 x 6) and racks

supplies:
system for

production

of pure water;

(120 x 80 cm, 80 cm high).

filtration

and distribution

1 Biohazard vertical
laminar
clearance
over worktop;

airflow

of media:
cabinet

for

two people,

with

at least

75 cm

WHO/Rabies/90.201
page 10

Rev.1

2 stainless

steel,

pressurized

1 pressurized
filter
filtered
compressed

filter

canisters,

20 litres

each;

canister
(5 litres),
also stainless
steel
pure anhydrous nitrogen);
air or, better,

(pressurized

2 stainless
connectors;

steel,

flat

disc

filters

(293 mm diameter),

with

joints

and

2 stainless
connectors;

steel,

flat

disc

filters

(142 mm diameter),

with

joints

and

4 magnetic

stirrers

(2 large

by

and 2 small);

1 pH meter;
1 water

bath

(20 to 6OyC), with

1 refrigerator
1 metering
1.4.2~

Cell

(+2" to +8"C),
pump to fill

culture

thermostat,

30 litres

capacity;

of 100 litres;

containers

(adjustable

between

10 ml and 1 litre).

room:

1 Biohazard vertical
laminar airflow
cabinet for four workers, with
Alternatively,
a laminar ceiling
75 cm clearance
above the worktop.
used in the centre of the room (working surface of 200 x 150 cm);

at least
could be

on a Bellco
1 roller
system: see figure
8, page 46, showing a set of rollers
(e.g. Bellco rollers,
2 units of 9 modules each, for 180 bottles
of 2 litres
- 76 x 62 cm, 2.5 m high; or 3 units of 6 modules each, 1.75 m high);
1 standard

microscope;

1 inverted

microscope;

1 pH metre;
1 shaker
1 water

for

bath with

2 magnetic
1 table

homogenization
thermostat,

top,

refrigerated

1 refrigerator

airflow

exchanger,

1 pH meter;
stirrer.

4 x 300 ml;

200 kg;

100 litres.

is transferred

cabinet

6000 rpm, capacity

capacity

(+2 to +8"C),

1 temperature

1 magnetic

30 litres;

centrifuge,

balance,

Room to which virus

1 vertical

suspensions;

shakers;

1 Dyana electronic

1.4.2d

of cell

for

after

inactivation:

one.worker;

circulation

principle,

with

thermostat;

unit
each

WHO/Rabies/90.201

1.4.2e

Room for

final

preparation

1 vertical

airflow

1 magnetic

stirrer.

1.4.3

Rev.1
page 11

of vaccine:

cabinet

for

two workers;

Area 3
Room for

cultures

and harvest

of virus:

1 Biohazard vertical
laminar airflow
cabinet
replaced with a central
laminar ceiling;
1 peristaltic

for

four

It

people.

could

be

pump;

1 metering

pump, as described

1 standard

microscope

1 inverted

microscope;

in 1.4.2b;

equipped

for

fluorescence;

1 pH meter;
1 water

bath with

4 magnetic

thermostat;

stirrers

(2 small

1 Dyana electronic
1 refrigerated
2 stainless
4 filter

balance,

steel

roller
units
modules) for

4000 rpm (6 x 1 litre

pressurized

cases for

1 stainless

filter
flat

disc

as described
300 two-litre

-15"

1 double
1.4.4

Various

entry

to -2O"C,
autoclave,

containers

filtration

for

filter,

Borosilicate

1.4.4~

Polypropylene

293 mm diameter,

in 1.4.2~
bottles;

(3 units

100 litres

joints
with

20 litres;
and connectors;

joints

and connectors;

of 9 modules or 5 units

equipped with cassettes

speed volumetric
pump;

(cut-off

point

of 6

at 100 000) with

capacity;

as in 1.4.la.
all

areas

Pyrex glassware
or teflon

capacity

of 0.753 m, with

1.4.4a 12 stainless
steel drums, 30 litres
capacity,
steel screw tops and 3 pipes, with inlet
at the top,
supply.
1.4.4b

capacity);

canisters,

cartridges

1 ultrafiltration
device
connections
and variable
1 freezer,

200 kg capacity;

centrifuge,
steel

and 2 large);

flasks

with wide necks and stainless

outlet
at the bottom, and air

between 5 ml and 15 litres

with

airtight

lids

for

capacity.
storage

of viral

seeds.

WHO/Rabies/90.201
page 12

Rev.1

1.4.4d Airtight
cryotubes
of 2 and 5 ml for storage of cell
or sealable phials of glass that can withstand
temperatures
1.4.5

Miscellaneous
Glassware

1.5.

seed in liquid
nitrogen,
down to -180C.

and other

small

pieces

of equipment

used in virology

STAFF (Number and qualifications)

1.5.1
In accordance with the general manufacturing
requirements
(WHO Technical
Report Series, No. 658, 1981, page 103, paragraph 2), rabies vaccine shall be
produced by staff who do not have to handle any other infectious
agents during the
The health of the staff
should be verified
and certified.
All those
production.
working in the sectors of vaccine production
and control
must be immunized and have
an antibody titre
of at least 0.5 IU per ml of serum.
Those who are not involved
in
production
or control
shall not have access to the sterile
working areas.
1.5.2
The qualifications
of those responsible
for production
and control,
should be forwarded to the national
authority
for the signing of protocols,
approval.
1.5.3

The production

staff

will

comprise

and thus
for

at least:

1 senior scientist
(the unit chief)
with a postgraduate
diploma in biology
followed by specialist
research and practical
experience
in microbiology
and
He will
also have trained
in a rabies laboratory
(preparation
of
virology.
virus and cell cultures,
analysis
and control);
1 highly
qualified
experience
in cell

technician
cultures,

with a degree in biology

virology
and microbiology;

and considerable

4 technicians
specially
trained
in cell culture
techniques,
working mainly in
It is preferable
to have two men and two women, since a
the sterile
areas.
number of delicate
operations
on cell cultures
are often better
done by women,
whereas other more strenuous tasks are more suitable
for men;
2 laboratory

staff

for

the non-sterile

area

(Area 1).

WHO/Rabies/90.201
CHAPTER 2.
2.1

Rev.1
page 13

CONTROLS

INTRODUCTION

2.1.1
A number of the vaccine controls
of cell cultures
naked eye and microscope examination
of
production
unit:
after infection
with the virus,
and of "control"
cells.
incubation
of the filtered
media before their use is also

must be performed in the

cell cultures
before and
Sterility
control
by
performed in situ.

2.1.2
With those exceptions,
all controls
of vaccine under preparation
and of the
finished
product must be made in a unit which is separate from the production
unit.
NOTE:

The control

unit

must also

be able

to control

the raw materials.

2.1.3
The laboratory
plans, the qualifications
of the staff,
and the control
must receive prior approval from the national
control
authority.
protocols,
2.1.4
When several vaccines are manufactured
on the same site,
the laboratory
controls
of all the vaccines
Chapter 1 (1.1.4),
and the same building.
2.1.5
The animal controls
must be conducted
described
in Chapter 4, animal houses.
2.1.6
All the controls
(in vitro
tests,
production
site)
come under the authority
the relevant
national
authority.
2.2
2.2.1

in separate

as described
in
can take place in one

installations.

animal testing
and controls
of the scientific
officer

These are
on the
responsible

to

BUILDINGS
Control

building

2.2.la
As in the case of the production
buildings,
there is a choice to be made
between a purpose-built
construction
and one made of prefabricated
elements or
modules set together.
2.2.lb
There is no need for sterile
areas.
The unit is designed
air-conditioned
building,
of the same materials
as the production
equally smooth and easily disinfected
floors,
walls and ceilings.
2.2.1~ In the sterile
operations
room, either
cabinet can be used for protection.

a Bunsen burner

as an
buildings,

with

or a laminar

airflow

2.2.ld
The building
is not necessarily
supplied with steam, nor does it need a
washing plant or sterilization
equipment, since the small volumes that need to be
decontaminated
can be treated with antiseptics.
After use, the recipients
and other
pieces of equipment are placed in air-tight
containers
and sent to the central
The washing plant also provides
the control
laboratories
with sterile
washing plant.
Ready-to-use
media can be supplied by the central
unit for media
equipment.
preparation.
2.2.2

Layout

of the control

2.2.2a The plan of the control

for the laboratory.

laboratorv
unit,

plan no. 3 on page 47, shows one possible

layout

WHO/Rabies/90.201
page 14

Rev.1

all

rooms open on a central

2.2.2b In this
sectors:

example,

1 sterility

control

laboratory

(testing

1 bacterial

control

laboratory

(2 rooms);

1 viral

vaccine

1 laboratory

control

for

laboratory

bacteria,

There are several

fungi

and mycoplasmas);

(3 rooms);

chemical

physical,

for

corridor.

and'biochemical

analyses.

The corridor
also provides
access to an incubation
chamber with two compartments
(the temperatures
of which can be regulated
separately)
a cold room, a room for
decontamination
and for storage of air-tight
containers
holding
equipment to be put
and a room for storage of samples.
through the autoclave,
On one side of the entrance to the building
are the offices
a double air lock in which the staff
change their clothes before
area.

and on the other is

going into the work

2.2.2~ In each section the size of the working surfaces are calculated
from the
volume of control
activities
to be undertaken,
which in its turn depends on the range
of vaccines to be controlled
and the number of lots of each vaccine received.
2.2.2d As regards viral
to have at least three
- one room for

vaccines,
irrespective
of the volumes
non-communicating
control
rooms:

cell

- two rooms for

cultures

tests

into

involving

which no viral
virulent

strain

concerned,

it

is

is brought;

material.

Such a division
makes for flexible
working arrangements
and safe control,
avoiding
the situation
where several strains
of different
viruses
are handled
same room in the same day.
2.3

is provided

with

the basic

Each room has work benches supplied

hot and cold water.

2.3.3
The building
equivalent
machine
2.3.4
airflow

in the

EQUIPMENT

2.3.1
Each section
concerned.
2.3.2
sink,

better

with

equipment

gas, compressed

has a demineralized
water supply
to produce pure, sterile
water.

Each room used for cell

cabinet with space for

2.3.5
The control
laboratory
vaccine for veterinary
use.

needed for

culture
and virology
two workers.

as well

air,

has a Biohazard

necessary

for

control

Controls
require,
in addition
to the usual
equipment and raw materials,
pages 28-38):

equipment

for

virology

2 adjustable

electricity,

as a "Milli-Qn

has the equipment

1 immunofluorence

the discipline

vertical

compartments

- one at 37"C,

or
laminar

of rabies
(see list

microscope;

incubation

one at 33C (+ 0.5");

of

WHO/Rabies/90.201

in the chemistry
and biochemistry
section:
with
a multiscan
photometer
to measure viral
well
as a spectrophotometer
that
can register
2.3.6
It is
sedimentation
of qualitative

equipment
for
nucleocapside
ultraviolet

immunoenzyme
tests
and glycoprotein,
light.

worth
acquiring
a CO2 incubator
and an ultracentrifuge
of rabies
virus
and for saccharose
gradient
separation
analysis
of the viral
antigen
obtained.

sufficient
space must be reserved
2.3.7
Finally,
taken at different
stages
of production.
Vertical
The sample room should
contain
for this
purpose.
+4"C, 1 at -30C and 1 at -80C.

for
in

the

in the sample room for

cabinets
with
drawers
at least
three
cabinets:

Samples of rabies
virus
before
inactivation
are preserved
inactivated
virus
and the final
adjuvanted
vaccine
are kept at
2.4.

Rev.1
page 15

interests

samples
are the best
1 at

at -80C;
+4"C.

the

STAFF

2.4.1
Staffing
performed.

levels

are

2.4.2
The person
responsible
and considerable
experience

determined

in

by the

total

for these controls

quality
control

of

volume

of

controls

should
be a scientist
biological
products.

to be

with

a degree

2.4.3
This person
should
have two assistants,
also scientists
with
experience
in
usual
control
techniques
employed
in bacteriology,
virology,
chemistry
and
biochemistry.
At least
one of them should have done some work in a rabies
vaccine
control
laboratory.
2.4.4
The specific
controls
of rabies
vaccine
can be conducted
by two part-time
technicians.
One must have training
in analysis
techniques
(both chemical
and
biochemical),
the other
in cell
and virus
cultures.
2.4.5
tested

as

These staff
at regular

members
intervals

should
be vaccinated
(antibody
titre
of

against
at least

rabies
and serologically
0.5 IU per ml of serum).

the

WHO/Rabies/90.201
page 16

Rev.1
CHAPTER 3.

3.1

CENTRAL SERVICES

INTRODUCTION

These services
offer facilities
for every unit in the production
centre.
They
are therefore
of great importance.
Their design, installation
and equipment must be
carefully
studied and selected,
since faults
and shortcominge
jeopardize
the
objectives.
These services
are:
a central

washing

plant;

a media preparation
a distribution
a central

service;

and packaging

service

that

stores

and distributes

the vaccines;

store;

the general

administration;

the essential/common
services.
This is the heart of the unit,
running not only of the production
and control
units but also
central
sesvices.
3.2
3.2.1

which sees to the

of the rest of the

CENTRAL WASHING PLANT

Specifications

3.2.la
It is important
to plan for space that is big enough for the washing and
sterilization
of equipment from the production
and control
units.
This area should
include space for equipment waiting
to be washed, and space for storage after
A rule of thumb is that the total
area of a washing plant should be
sterilization.
approximately
a fifth
of that of the laboratories
served.
Thus on the production
site
illus
rated on plan no. 1, page 39, the planned surface area of the washing plant is
390 m5 , one fifth
of the surface of the buildings
located under the central
hangar.
3.2.lb
This approximation
equipment passing through
3.2.1~

The washing
a room for
a soaking

will have to be amended in accordance

the washing plant.

plant

(plan

reception

of soiled

and washing

an assembly
a room for

main parts:

equipment;

of clean

room;
equipment.

Each of these rooms must be big

equipment to circulate
freely.
3.2.2

of four

the volume of

room;

and sterilization
storage

no. 4, page 48) consists

with

enough to allow

the trolleys

and vehicles

carrying

Rauinment

3.2.2a The washing plant is supplied with electricity,

steam, deionized water, and if
It also needs distilled
water.
necessary gas, by the general services.
It is
therefore
often worth equipping
it with single or double distillation
apparatus,
so
that water can be used directly
from the machines or stored in stainless
steel
containers
at a temperature
of 80C.

WHO/Rabies/90.201

Rev.1
page 17

3.2.2b The washing room is equipped with tubs either made of stainless
steel or lined
Some are used for soaking the equipment in
heat-resistant
plastic.
with inert,
detergent,
others for scrubbing and rinsing
with demineralized
water and distilled
The room also has automatic pipette
washers.
water.
An automatic washing machine with several programmes might be considered,
when
there is a large quantity
of equipment including
bulky objects such as large glass
in countries
where labour costs are high.
bottles
and stainless
steel drums, especially
3.2.2~ The sterilization
room contains
a large rack for preparation
of the washed
which involves
several operations:
putting
the
equipment before sterilization,
seals in aluminium
pipettes
in cylinders,
putting
stoppers in boxes, wrapping bottle
foil,
assembling of tubing and connecting
systems, assembling of filters
etc.
Sterilization
is then done either by autoclave
(drums, flexible
tubes, stoppers,
filters
etc.)
or by dry heat in an electric
oven (glassware).
Two autoclaves
of
The autoclaves
and the oven have the same
different
sizes offer greater flexibility.
specifications
as those in the building
for production
of rabies vaccine (paragraph
1.4.1.a).
3.2.2d

Special

notes

on washing

of equipment

from rabies

vaccine

unit.

The equipment that has been used for cell and virus cultures
is decontaminated
in an autoclave before being sent to the washing plant.
Proteins
and cell debris
Therefore,
they must be
coaguate on the walls of vessels which have been autoclaved.
soaked in tubs containing
an appropriate
detergent
solution
(such as 7X) or be
Disposable
treated with trypsine
solution
before they can be properly
washed.
plastic
bottles
for cultures
(see list
of equipment and raw materials
pages 38-48),
offer the best material
for cells to anchor on and considerably
reduce the quantity
of equipment passing through the washing plant in the process of vaccine production.
For cells to grow and multiply
in vitro
and survive
in good physiological
it is essential
that the equipment used is perfectly
clean.
condition,
After
be rinsed once with an acid solution,
several
washing, the glassware should therefore
times with abundant, deionized water, and finally
with distilled
or double distilled
water.
Excellentresults
these
and rinsing;
3.2.3
units,

are produced by machines that are purpose built

for
range in size from laboratory
to industrial
models.

washing

Staff
The team leader
and supervises

organizes
and plans
operations;

the work in the light

of requests

from other

The size of the team working under the leader is dictated

by the workload,
reception
of equipment,
soaking and washing,
divides
into several occupations:
sterilization,
assembly and packaging.

which

It is always good to have staff capable of doing more than one job, and it
especially
useful to have at least two people who are well-versed
and practised
use of the machines (washing machines, autoclaves,
ovens, etc.).

is
in

3.3.

MEDIA PREPARATIONUNIT

3.3.1

Snecifications

3.3.la

The media preparation

unit

is considerably

smallyer

than the washing

plant,

WHQ/Rabies/90.201
page 18

Rev.1

"standard"
buildings
of the-production
and can be housed in one of-the
centre.
In the arrangement shown in plan no. 1 on page 39, it is part
extension
of the viral
vaccine unit,' near the central
washing plant.
3.3.lb
The premises for
no. 5, Media preparation

preparation
of cultures
unit,
page 49):

consist

of several

and control
of the

rooms (see plan

an office:
a store
a cold

room for

raw materials;

room;

an incubation

room (temperature

a room for

weighing,

a room for

filtration

mixing

37C + 0.5");

and preparation

of solutions;

and filling.

It has
The filtration
room is a protected
area (shaded in plan no. 5, page 49).
a supply of filtered
air (through absolute filters)
and is kept at higher than
Staff members gain access through an airlock,
where they don
atmospheric
pressure.
protective
clothing.
The store room contains
all raw materials
except those that must be kept at a'
The raw materials
that can be checked must be
temperature
between +2" and +8"C.
given a clearly
visible
label to show that they have passed inspection.
ready
for

The cold room is for

for use.

The incubation
inspection.

3.3.1~
plant.

Media that

the storage

room is for

both

incubation

of heat

sensitive

of filtered

are to be heat-sterilized

raw materials

media before

are autoclaved

3.3.2

samples are taken

at the central

3.3.ld
Culture media to be used in preparation
of rabies vaccine
should if possible
be sent unfiltered
to the production
building,
sterilization
is carried
out.

and media

washing

on cell cultures
where the

Rauioment
The main items

of equipment

are:

temperature

+2'

1 refrigerator,
1 freezer,

temperature

-15"

magnetic
1 water
litres;

capacity

to -2O"C, capacity

with
several filtration
units,
293 mm and 142 mm in diameter,
several stainless
20 litres;

to +8'C,

200 litres;

200 litres;

supports, for stainless

steel,
with joints
and connectors;

steel,

pressurized

heating

to between

filtration

canisters,

flat

disc

capacity

filters

2 litres

stirrers;
bath

for

20' and 6O'C, thermostated,

capacity

30

and

,-

WHO/Rabies/90.201

heating

capacity

vats,

2 pH metres

(accuracy

1 stainless

steel

and 120 litres;

of + 0.05 pH);

mixing

balances of various
of + l/10 mg);
1 peristaltic

40 litres

Rev.1
page 19

vat

(capacity

capacities

150 litres)

with

turbine;

200 g and one precision

(10 kg,

balance

(accuracy

pump;

2 trolleys.
In the sterile

area where filtration

a horizontal
apparatus

laminar
with

is conducted:

airflow

a metered

cabinet

pump for

for

2 workers;

distribution

of filtered

Staff

3.3.3

3.3.3a The number of people preparing

media can be halved if
these contain most of the relevant
powdered media are used;
advantage of constant quality.
3.3.3b This unit must be run by a scientist
preparation
of media.
3.3.3~

The subordinate

technicians

in large-scale
in situ

if

possible,

the number of interchangeable

staff
required
for
every 2000 litres
of media and reagents to be

Introduction

This unit fills

all the inactivated
Live vaccines should
for distribution.
or in a separate room within
the unit.
within

trained,

available
and have the

FILLING AND PACKAGINGUNIT

3.4.1

I-.

commercially
ingredients

who is experienced

must be specially

A rule of thumb for calculating

the various posts is one person for
produced each year.
3.4

media.

vaccines produced on the site in containers

be packed at the end of the production
line

The filling
unit must ensure that the products are free
or without;
the environment must therefore
be sterile

3.4.2

Soecifications

The sterile
and kept

of this building
are the same as for
(see Chapter 1, paragraph 1.3.2).

area is supplied with filtered

and conditioned
at higher than atmospheric
pressure.

air.

building
It

As in the rabies building,

the area is entered through double airlocks
women) where the staff
dons protective
clothing
(ready for them in sterile
All

surfaces

from

of the buildinz

3.4.2a The general specifications

for production
of rabies vaccine
3.4.2b
airtight

of contamination
and protected.

must be smooth,

dust free

and easily

disinfected.

lE,

must be
(for men an
wrapping).

WHO/Rabies/90.201
page 20

Rev.1

Apparatus for disinfection

treatment
of each room.
The rooms are supplied

with
with

3.4.2~ The building

has neither
near the central
washing plant,
3.4.3

formalin

vapour

electricity,

washing plant nor sterilization

which provides
sterile
material

separate
air.

equipment,
in air-tight

since it is
containers

Rooms and Annexes

In the same building,

on either

side

(plan no. 6, page 50) shows the

the sterile
area, where access to

of the sterile

area,

are:

an office
and a cold room for products
in bulk
At the entrance,
and access to the airlock
into the sterile
area;
packaging,
At the air-locked
exit
are packed for delivery.

from the sterile

3.4.3~ On one side of that room is a cold

bay for the finished
products.
3.4.3d The size of the premises
In dealing with
to be packaged.
annum are made from a series of
form of individual,
1 ml vials,
in mass vaccination
campaigns).
minimize loss of vaccine in use,
packaged are doubled.
3.4.3e

for

gas, oxygen and compressed

3.4.3a The plan of the filling

and packaging unit
lay-out
of the filling
and packaging rooms within
the rooms is from a side corridor.
3.4.38

must be available

Rooms in the sterile

area,
store

awaiting

a room where the final

room, and on the other

products

is a loading

should be determined by the total volume of products

rabies vaccine for veterinary
use, 500 000 doses per
20 batches of 25 litres
each and presented
in the
or multidose
bottles
(the latter
should be used &
At times 2 ml doses are preferred
in order to
and in such cases the quantities
of vaccine to be

area

at least two rooms should be provided so

If there is a central
packaging unit,
For safety's
sake, the
that different
vaccines can be bottled
at the same time.
entrance to these rooms should be a double-doored
airlock.
containing
Beyond the bottling
rooms is an inspection
room. Vials and bottles
Only
cloudy suspensions
in liquid
such as rabies vaccine are inspected visually.
products in transparent
solution
should be inspected by sorting
machines, which are
expensive.
The labelling
and packaging room is the last room in the sterile
can be done either by hand or by simple or sophisticated
machinery.
operations
to be conducted in this room can be reduced if the filling
machinery that can also label the recipients.
3.4.3f
crates

There must be space in the packaging

in which the vaccine is delivered.

3.4.4

gauinment

3.4.4a

Bottling

room for

the storage

area.
The work
The number of
rooms have

of boxes,

cartons

rooms

These rooms contain machinery adapted to the type of containers

to be filled,
the type of sealing
to be used, the unit volumes to be prepared as well as to the
working conditions.

or

WHO/Rabies/90.201

Rev.1
page 21

The supply of sterile

air to those rooms should enable vaccines
to be bottled
in
Sterile
air, once it has passed through absolute filters,
can
complete asepsis.
therefore
be circulated
in the rooms through conventional
vents, although a vertical
above and around the bottling
machine is a more effective
method.
laminar air "tent"
Another less complicated
method is that of placing
the machines under Biohazard
airflow
cabinets placed on rollers
and equipped with removable lateral
flaps.
The bottling
room should also
containers
shakers, double-skinned
3.4.4b

Optical
This

3.4.4~

inspection

contains

Packaging
This

contain small pieces

for the refrigeration

of equipment, eg. magnetic

of heat sensitive
vaccines,

et

room:

a magnifying

viewer

and an automatic

sorting

machine

if

necessary.

room:

contains

equipment

for

hand-labelling,

and perhaps

for

automatic

labelling

also.
-.

3.4.4d

Packing

room:

If the packing is done by hand, a large table is sufficient.

The cold room next
door should have separate sets of shelves,
coded for easy identification,
so that
stocks of different
products are not mixed up.
3.4.4e Trolleys
must be available:
at least one at the entrance,
area and one in the packaging and despatch area.
3.4.5

two in the sterile

Staff

The number of staff

employed depends on the annual volume of work.
In a small
team, employees should preferably
be able to do more than one of the jobs involved.
A team leader supervises
all activities
and monitors
stocks of vaccine.
It
good if the team leader or another employee of the unit is able to make routine
adjustments
to the machinery.
3.5
3.5.1

is

OTHER GENERAL SERVICES

Uenot

Premises that are separate (as in plan no. 1 page 39) or incorporated
in one of
the buildings
must be set aside for reception
and storage of raw materials,
small
and maintenance of heavy plant.
equipment, and the main spare parts needed for repair
Arrangements
unit

must also be made for

some products

at low temperature.

Un-inspected
goods must be kept separate from goods passed by the inspection
of the facility
and labelled
with inspection
codes and numbers.

A facility
producing
manage the depot.
3.5.2

keeping

several

types

building
or noises.

should

of vaccine

should

have at least

two people

Administration

The administration
from unpleasant
smells

be near the site

entrance

if

possible,

away

to

WHO/Rabies/90.201
page 22

Rev.1

It should have enough offices

for the management, the secretariat,
the accounts
The archives
should include all records of the main data
and archives
departments.
A record should
and results
in accordance with production
and inspection
protocols.
Standard records for rabies
be kept for each batch of every vaccine manufactured.
vaccine for veterinary
use are presented in WHOTechnical
Report Series, No. 658,
1981, Annex 3.
The administration
facilities
should also include a meeting
and a medical service containing
an infirmary
double as library,
emergencies and a surgery for physicians'
visits.

room that could

equipped for

a canteen should be provided,

if there are no external
food services,
Finally,
since the basic rules of hygiene and safety forbid
food in the laboratories
and
annexes.
3.5.3

Essential

services

3.5.3a This is the heart

energy, water, ventilated
emergency repairs.

of the establishment,
providing
as well
air, heat, cold, etc.,

the crucial
supplies
of
as equipment maintenance

and

3.5.3b A detailed
study of the requirements
of the essential
services
must, be made
It is recommended that capacity
be
as soon as the production
targets
are known.
increased immediately
if increased production
or the creation
of new units is planned
in the short- or medium-term.
In view of the special
skills
involved,,
these needs should be estimated,
the
machinery and equipment chosen and the plans for the centre and distribution
networks
drawn up in close collaboration
with specialist
engineers
for each of those areas.
3.5.3~

The main activities

of this

department

fall

under

the following

headings:

surely.
This usually
comes from an external,
high-tension
power
Electricitv
Power is transmitted
to
there is a transformer
at the entrance to the site.
line;
the other buildings
from the central
control
unit of the central
building
of
In each building
power lines for all rooms emanate from
essential
services.
Appropriate
safety devices must be
protected
cupboards with triple-phased
current.
installed
in each one.

The service must be prepared to cope with any power failure.

The essential
services building
should contain several generators
with enough power to supply the
culture
apparatus and animal houses.
The
incubation
rooms, cold rooms, freezers,
back-up generators
require
a nearby oil.supply.
Steam SUDD~Y. A group of boilers
in the building
especially
for sterilization
of equipment (autoclaves,

provides
the necessary
or in situ fermenters).

Water SUDD~V. Ordinary and deionized water, the latter

ion-exchangers,
should also be supplied by the'main building
services.

steam

produced through
of the essential

Ventilation.
Because of the complexity
of the ventilation
equipment, only
specialist
technicians
and engineers are able to recommend the most appropriate
systems according
to climatic
conditions,
volume of the premises and special
features
of the various
laboratories,
including
the sterile
areas that must be supplied with
air filtered
through independent circuits.
Waste disposal.
incinerated.

Waste water

drains

to a sewage, and solid

wastes

are

WHO/Rabies/90.201

Rev.1
page 23

3.5.3d
The staff
of the essential
services
unit
carries
out the above tasks
and
The technicians
are also responsible
for emergency
repairs
maintains
the plant.
for manufacturing
certain
light
apparatus
and various
tools.
The unit
therefore
needs workshops
(number 10 on the plan for a biologicals
production
unit
- plan
no. 1, page 39).
The head of unit
must
by one or more maintenance
Essential
electro-technicians,
and carpenters.
They
possible.

.-

should

services

be a highly
engineers.

qualified

and experienced

also need a team consisting

mechanics,
heating
engineers,

be trained

in

fire-fighting

engineer,

and

assisted

of one or more electricians,

refrigeration
engineers,
plumbers

techniques,

at

the

work

place

if

WHO/Rabies/90.201
page 24

Rev.1
CHAPTER4:

4.1

ANIMAL HOUSES

INTRODUCTION
Control

tests

performed

on animals

come under

the central

control

unit

(see

2.1.6).
However, these tests are conducted in animal houses that must be at a physical
in a separate building
if possible,
away from the
distance
from the laboratories,
(blocks 14a and 14b in the overall
plan - plan no. 1 production
and control
centre,
on page 39).
The animal
4.2

handlers

should

not enter

the laboratories.

CENTRAL ANIMAL HOUSESFOR SMALL LABORATORYANIMALS (see eg, plan

no. 7, page 51)

4.2.1
The control
tests call for the use of mice, guinea pigs, rabbits
and other
species which are either bought as needs arise or bred at the centre;
numbers and
species are determined by the vaccines produced.
The choice is governed by local
resources and by cost.
4.2.2
For each species of animal, separate rooms should be devoted to (a) breeding
(if necessary),
(b) storage of animals prior to use and (c) observation
of animals
under inspection.
4.2.3
The animal houses should be ventilated
so as to renew the air 10 to 15 times
per hour, keeping relative
humidity
between 40% and 65% and atmospheric
temperature
between 18" and 24C.
4.2.4
soiled
4.2.5
found
4.3
4.3.1

There should also be an annex large

and for storage of clean
equipment,

enough for
equipment,

washing and sterilization

feed and bedding.

of

Guidelines
on design, equipping and running of such anima'l ho'uses are to be
in WHOManual BLG/UNDP/78.1, on production
and control
of bacterial
vaccines.
ANIMAL HOUSES FOR TESTING OF RABIES VACCINE FOR VETERINARY USE
Tests

on animals

4.3.la
The rabies vaccine tests
No. 658, 1981;
they include:
a test
tests

for
for

are described

the absence of abnormal

safety

(inactivation

a potency test on mice using

with rabies virus.

in WHOTechnical

toxicity,

on mice and guinea

and innocuity)
the NIH test,

4.3.lb
The same technique will be used for
Series, No. 790, 1983, paragraph 5.4).

Report

Series,
pigs;

on mice;
by vaccination

stability

tests

followed
(WHO Technical

by challenge
Report

of the vaccine on the

4.3.1~ It is also necessary to evaluate the immunogenicity
followed by blood testing
to ascertain
target species - the dog - by vaccination
It is also important
to estimate the duration
of
titres
of specific
antibodies.
protection
conferred
by a newly manufactured vaccine on the first
batches produced.
dogs and control
animals must be kept for one, two and
For this purpose, vaccinated
even three years before they are challenged with a potent wild virus strain.

WHO/Rabies/90.201

4.3.2.

Specifications

of the animal

houses used for

testing

of rabies

Rev.1
page 25

vaccine

4.3.2a In view of the risk of contamination

with rabies vaccine,
the only test
can be conducted in the central
animal house is the test for abnormal toxicity
vaccine produced.

that
of the

4.3.2b All tests involving

inoculation
of the virus-vaccine
before inactivation,
and/or challenge
strains
must be performed in separate parts of the central
animal
house used for that purpose alone (see plan no. 1 of a centre for biologicals
production,
page 39).
4.3.2~ Two types
veterinary
use:

of animal house are needed for

housing for mice and kennels.

testing

of rabies

vaccine

for

4.3.2d The building

and running of kennels equipped for performing
challenges
with a
wild strain
of rabies vaccine puts a heavy burden on the production
process.
Tests
on dogs in general are only conducted with a few batches of vaccine,
at the start of
such tests show what protection
is given to the species concerned by the
production;
vaccine,
which is produced in accordance with a finalized
manufacturing
protocol.
Each country should therefore
explore the possibility
of conducting
such tests in a
national
or foreign
establishment
equipped with an isolation
animal house that has
the proper safeguards and accepts the responsibility
(e.g. a regional
centre for
rabies studies).
4.3.2e If testing
on dogs is restricted
to vaccination
and testing
of blood samples
a conventional
kennel for 15 to 20 dogs is sufficient.
The
for titres
of antibodies,
animals can be placed in cages in a closed room or in niches opening on to a cemented
o the kennels are a store room for equipment
area surrounded by wire mesh. Adjacent
and food and a room of approximately
8 m5 with a table for restraining
animals
while blood samples are taken.
4.3.2f
If, on the other hand, a kennel for performing
challenges
on dogs is needed,
it must be installed
in an enclosed building,
as an isolated
area in airtight
conditions
similar
to those in the building
where rabies vaccine is produced from
For practical
reasons it is worth installing
the animal house for
cell cultures.
tests on mice in the same building,
in a second area separate from the kennels,
without
the same degree of protection
(unless a wild strain
of virus is being used).
Plan no. 8 on page 52 shows one arrangement for such facilities
in an animal house
for the testing
of rabies vaccine on mice and dogs.
height,
"wild"

the building
itself
is surrounded by a wall some 4 m in
To increase security,
with two sliding
doors for an entrance.
It must be designed to prevent
animals (especially
small rodents)
from going in.

4.3.3

Housine

4.3.3a

This

for

mice

comprises:

an entrance corridor
with airlock
where the staff
shoes.
New mice, equipment and feed are brought
a storeroom

for

put on working clothes and

in through the same airlock;

feed and bedding;

two rooms where mice are kept on shelves in boxes of 5 and 10, both with access
to an inoculation
box.
The first
room is for vaccinations,
the second for
challenges.
There is no place to keep mice in readiness:
they must be brought
in just before they are used;

WHO/Rabies/90.201
page 26

Rev.1

at the exit an airlock

for decontamination,
using formalin
or another active
process, where discarded bedding, waste or dead animals are kept in bags before
incineration.
Equipment is immersed in an antiseptic
solution
of formalin
or
bleach before being sent to the washing plant.
4.3.3b To test the annual production
of the 20 batches of vaccine assumed in these
Instructions
(and irrespective
of the size of the batches),
the animal house should
be able to accommodate 300 mice in 60 cages at any one time.
Cages consisting
of a
feed tray and drinking
water, that
plastic
container
with a stainless
steel mesh lid,
can be sterilized
in an autoclave
(see figure
9, page 53) are easiest
to use.
4.3.3~ The animal house should be ventilated
by maintaining
higher than atmospheric
air pressure,
the outgoing air being decontaminated
by passage through a conduit
where the temperature
is raised to 180C (or any other active decontamination
process.
4.3.4

Isolation

kennel

for

vaccination

followed

It
isolated.
It is airtight,

4.3.4a The kennels and annexes must be strictly

protected
animal house in plan no. 8, page 52.
adjusted to produce negative pressure.
Extracted
Used water
decontamination

air

must be sterilized

(especially
vat before

by burning

water used for

expulsion.

These safety measures are justified

of the virus used for challenge.
4.3.4b

The kennels

by challenge
takes the form of a
and ventilation
is

in a conduit

washing

at 180C.

the kennel)

by the high

virulence

must pass through

of challenge

strains

comprise:

an airlock
protective

entrance
clothing;

for

staff

with

shower and room for

an airlock

entrance

for

clean

equipment,

food,

dressing

and experimental

in full
animals;

the animal house, with a central

corridor
and floors
that slope to drain off the
The dogs are kept
water used for cleaning,
and several pressurized
water taps.
on either
side of the corridor
in individual
kennels of metal (see figures
10
and 11, page 54);
one laboratory
equipped with a biohazard
the central
passageway with openings:
specimens of
vertical
airflow
cabinet for one person, who prepares the challenge
one room with a table on which the dogs are restrained
during
rabies virus;
a spare room for equipment and food;
inoculation
and taking of blood samples;
-

a room at the exit with a double-entry

autoclave
(capacity
1 to 2 m3) for
decontamination
of equipment and clothing
which is also used for the
sterilization
of solid wastes and dead animals which arrive
in plastic
bin bags.

at the outer door of the autoclave,

sterilized
products
4.3.4~ Outside the kennels,
are taken through a special
airlock
where the outside of the wrappings are
this airlock
is also used for the mouse housing.
Once they have
decontaminated;
the disposal bags and the dead animals are
been taken outside the surrounding
wall,
incinerated,
and the equipment is sent to the washing plant.

WRO/Rabies/90.201

4.3.5

Staff

4.3.5a Staff
neutralizing
4.3.5b If
people.
4.3.5~
animal

,-.

Rev.1
page 27

must be vaccinated
antibodies
should

against rabies and their levels

be checked every 8 to 12 months.

the mice and dogs are kept

in the same area,

the unit

of specific
can be run by two

At least one of the handlers should have completed a training

session
testing
centre,
preferably
in a unit that tests for or studies canine

in an
rabies.

WRO/Rabies/90.201
page 28

Rev.1

ANNEX1
List

This
supplies.

list

features

of eauinment and materials

control
of rabies vaccine

only

the main plant

used in Droduction
for veterinarv
use

and equipment,

and

and the most commonly used

Some names of manufacturers

or suppliers
are provided.
There are, of course,
other suppliers
and alternative
products.
The listing
does not necessarily
imply
special recommendation by WHO.
Prices,

where available,

are quoted

in French Francs

.../...

WHO/Rabies/90.201

A:

ITEM

1.

2.

3.

-\

APPROXIMATEUNIT PRICE
(1986 - less tax)
in French francs

Sterilization
- Autoclaves
(l-2 m3)
with automated programmes

AMSCO, USA'
OLSA, Italy
LEQUEUX, France

- Pasteur

LEQUEUX, France
HERAEUSGmbH, Germany

300 000

100 000

Washing
- Laboratory
machines

washing

Industrial

washing

HAM0 by Flobio France

distributors
GALIAY, Switzerland
machines

25 000

BETTER BUILT Machinery

Corp., USA

- Machines for washing

virus ampoules or bottles

ARENCOGmbH, Germany
STRUNCK & Co., Germany
STRUERS, Denmark

- Machines for washing

animal cages

BETTER BUILT Machinery

Corp., USA

Water Droduction
- Separate bed deionizers
(100 1. per hour)

WATCO, France

- Distillation

OLSA, Italy

apparatus

- "Milli
Q" Pure water
generators
(100 1. per hour)
4.

EOUIPMENT

SUPPLIER

ovens

100 000

MILLIPORE CORPORATION, USA

50 000

Filling
- Filling

machines

Rev.1
page 29

ARENCOGmbH, Germany
STRUNCK& Co., Germany
STRUERS, Denmark

- Printing

machines

HAPA A.G.

- Perifill
metering

sequential
pumps

BIOBLOCK Scientific,

France

18 000

WHO/Rabies/90.201
page 30

Rev.1

A:

Refrigeration
- Liquid nitrogen
containers
for storing
seeds, 40 1.
capacity
(for 180 vials
of 2 ml)

Societe AIR LIQUIDE,

BIOBU>CK Scientific,

- Freezers
(300 1.)

FRIGIDAIRE
KELVINATOR
(Flobio France
distributors)

- Freezers
(380 1.)

6.

APPROXIMATEUNIT PRICE
(1986 - less tax)
in French francs

SUPPLIER

ITEM

5.

EOUIPMENT (cont.)

-30C

France
France

4 000
-

FROID-LAB0 REVCO (Flobio France

distributors)

-85C

50 000
-

- Crushed ice machines

INCO-ZIEGRA, Germany

- Refrigerators
(200 1.)

FRIGIDAIRE
KELVINATOR
(Flobio France
distributors)

+2 - +8"C

6 000

15 000
4 000

Heating
- Laboratory
(110 1.)

incubators

ASSAB (Flobio
distributors)

France

6 000

38 000

- CO2 incubators

7.

- Thermostatic
baths
(20 1.) and thermostat

BIOBLOCK Scientific,
ROUCAIRE, France

- Double-hulled
stainless
steel tanks with heat
exchangers

ROUCAIRE, France

Laminar
cabinets

France

5 000

to

25 000
35 000

airflow

BIOHAZARD (0.3~) with

HEPA absolute filters
(UV, recycled
air and
pull-down
shutter)

FLOW G., Luc. Company, USA

GEIMAN Instrument,
Italy
E.S.I.,
France
A.D.S.,
France
N.E.U., France

30 000 (1 place)
40 000 (2 places)
100 000 (3 places)

WHO/Rabies/90.201

A:

ITEM

8.

EOUIPMENT (cont.)
APPROXIMATEUNIT PRICE
(1986 - less tax)
in French francs

SUPPLIER

Filtration
- Flat filters
with membranes SARTORIUS, France
and accessories
SARTORIUS:WERKE, Germany
142 mm diameter and
MILLIPORE, France
293 mm diameter

15 000 (142 mm)

20 000 (293 mm)

- Filter
holders with
accessori
s for cartridges
of 0.75 m5

PALL, France

30 000

- Device for
membranes

checking

SARTORIUS, France
SARTORIUS-WERKE,Germany

15 000

- Pressurized
filtration
canisters
20 1.
5 1.

SARTORIUS, France
SARTORIUS-WERKE,Germany

-.

9.

10.

10 000
8 000

Concentration
Ultrafiltration
device
with packs of membranes:
cut-off
point - 100 000

MILLIPORE, France

Variable
PumP

BIOBLOCK Scientific,

speed metering

80 000

France

5 000

Mixing

,T--

Stainless
steel
mixing vat (200 1.)
+ turbine
11.

Rev;1
page 31

LABO. MODERNE, France

12 000

Balances
- Universal
(3-4 kg,

balances,
accuracy + 0.1 g)

- Analytical
balances
(200 g, accuracy & 0.1 mg)
- DYANA industrial
balances
(150 kg, accuracy of + 1 g)

METTLER, Switzerland
SARTORIUS, France
SARTORIUS-WERKE, Germany

7 000
18 000
7 000

WHO/Rabies/90.201
page 32

Rev.1

A:

13.

Centrifuges
- Refrigerated
centrifuges
6000 rpm, 4x300 ml
6000 rpm, 6 x 1 1.

SIGMA,

- Ultracentrifuge

BECKMANInstrument

Inc.,

USA

130 000

BELLCO Equipment

System,

USA

25 000

stages

BELLCO Equipment

System,

USA

- Glass bottles

BELLCO Equipment

System,

USA

Cell

and virus

- Basic roller
(10 x 2 litre
- Extra

14.

cultures
system
bottles)

OLYMPUS, France

with

to

- Normal or
inverted
stand

ZEISS, Germany

- Equipment for
fluorescence

ZEISS, Germany

8 000
12 000

20 000

pH metres
- Traditional
metre
- pocket

16.

35 000
70 000

MicroscoDes
- Binoculars
3 lenses

15.

APPROXIMATEUNIT PRICE
(1986 - less tax)
in French francs

SUPPLIER

ITEM

12.

EOUIPMENT (cont.)

digital

pH

pH meter

BECKMANInstrument

Corp.,

BIOBLOCK Scientific,

USA

France

3 500
1 300

Immunoenzvme tests
- Titertek
equipment with
multiscan
photometer
and printer

FLOW Laboratory,

USA

50 000

WHO/Rabies/90.201

A:

17.

APPROXIMATEUNIT PRICE
(1986 - less tax)
in French francs

Homoeenizers
- Airtight
vibration
10 1. capacity

18.

EOUIPMENT (cont.)

SUPPLIER

ITEM

Animal

system

CHENAPA.d.,

Switzerland

caees

- Mice
IFFA-CREDO, France
- Dogs

Rev.1
page 33

WHO/Rabies/90.201
page 34

Rev.1

Miscellaneous

Light

19.

eauioment

Such equipment is made by many manufacturers

suppliers
of laboratory
equipment.
- Magnetic
- Vortex

and can be obtained

from the main

stirrers
stirrers

- Microplate
- Masterflex

stirrers
transfer

- 30-litre

peristaltic

stainless

steel

pumps

drums (beer barrel

type)

- Meker burners
- Hemostatic

clamps

- Screw-on

clamps

- Countering

20.

cells

- Electrically

operated

- Eppendorf

micropipettes

Glassware
CORNING GLASS WORKS, USA
JENAIR GLASSWERKSCHOTTU.G. FRG

Main Suppliers:
Pyrex quality

borosilicate

glass

- Traditional

cylindrical

bottles,

- Graduated
- Various
21.

pipettes

cylindrical
test

Autoclavable
- Cylindrical

bottles

tubes,

(capacity
of (capacity

pipettes

etc.

test

tubes,

- Polycarbonate
and polypropylene
for storage at -85C
screw top tubes

- Rhodorsil

silicon

22.

silicon

Disoosable
- Special
- Pipettes
- Sterile

for

stoppers

- Grooved polypropylene
- Flexible

of 1, 2, 5, 10 and 15-litres)

elastics
bottles,

- Airtight

from 2 ml to 500 ml)

bottles
samples

of various

tubing,

tubing

funnels

I,

etc.
with

(5,

airtight

screw-on

lids

10, 20 ml)

sizes

(24 to 61 mm diameter)

T and Y shaped

in several

diameters

nlastics

airtight

tubes

in sterile

for

storing

wrapping

wrapped cones for

- 96-hole microplates
- Disposable
syringes

with

(l-10'ml)

Eppendorf
lid

(sterile,

the seed immersed in liquid

pipettes

(sterile)
with

needles

of 1, 2 and 5 ml)

nitrogen

WHOjRabies/90.201

22.

Disnosable

nlastics

- Vacutainers

for

- Rhodonsil
- Sterile

blood

flexible
latex

samples

pipe

surgical

- "Pneumofilter"

(8 x 12 and 8 x 14 diameter)
gloves

disposable

- Autoclavable
- Bottles

(cont.)

air

filters

with

grooved

nozzles,

sterile

bin bags

for

cell

cultures

("TC"

specially

treated)

Main suppliers:
FALCON
CORNING
GREINER
- Flat

bottles

of 25, 75 and 150 cm2 surface

- Bottles
for rollers,
30 and 40 French
23.

Protective
Societe
- Overalls

850 cm2 surface

francs)

area

area (bulk

price

clothing
ADS, France
(plastified,

non-flammable)

- Shoes and boots

- Hoods, masks,

caps

- Goggles
(Cost

approximately

2000 French francs

for

each suit)

between

Rev.1
page 35

WHO/Rabies/90.201
page 36

Rev.1

MEDIA AND MAIN RAW MATERIALS

B:
1. Virus

and cell

culture

media

FLOW or GIBCO

Manufacturer:

Ouantitv

MEM medium powder to make 50 litres

Calf

foetus

serum

Newborn calf
FV bovine
Tryptose

serum

albumin

serum (powder)

Ph. broth

ner batch

Ouantitv

in 20

Annroximate
total nrice in
French Francs
(1986)

100.0 1.

2 000 1.

24 000

3.5 1.

70 1.

56 000

3.5 1.

70 1.

14 000

75.0 g

1 500 g

15 000

5.0 1.

100 1.

5 000

15 g.

300 g.

2. Trvosin
l/250
Sunnliers:
Soluble

FLOW-DIPCO-GIBCO, etc.
in PBS without

Ca-Mg

6 000

WHO/Eabies/90.201

3. Additives:

amino acids.

Manufacturer:

sugars.

mineral

salts

etc.

MERE
Annual
Cuantitv

oer batch

reauirements
Quantitv

in 20

ADDrOXhEite

total nrice in
French Franca
(1986)
2 kg

Tricine

40 kg

L-glutamine

500 g

10 kg

L-histidine

50 g
25 g
100 g

1 kg
500 g

L-alanine
Glycine

2 kg
150 kg

Saccharose
Glucose

2 kg

Sorbitol
Tween 80

4 kg
1 1.

Tween 20

1 1.

Sodium bicarbonate

5 kg

Calcium

chloride

Magnesium sulfate

1 kg
500 g

Magnesium chloride

500 g

Monosodium phosphate

1 kg
500 g

Disodium

phosphate

Sodium chloride
Trisodium

4.,Virus

Manufacturer:

16 000

20 kg

citrate

inactivation

approx.

2 kg

aeent
FLUKA

- Betapropiolactone
(to be kept at -20C)

300 ml

65 ml

13 000

5. Adluvant
Manufacturer:

SUPFXFOSSpeciality

- Alhydrogel

3.5 1.

Chemicals,

Denmark
70 1.

3 000

Rev.1
page 37

WHO/Rabies/90.201
page 38

Rev.1

Miscellaneous

6.

- 7 x detergent

for

soaking

containers

(FLOW)

- 90" alcohol
- Caustic

soda

- Hydrochloric

acid

- 40% formalin
- Ammonia etc.
Reagents

7.

for

- Oxygenated

immunoenzvme and immunofluorescence

tests

water

- 0-phenylendiamine
-

Antibodies

conjugated

Reference

Fluorescent

with

peroxydase
-

antigens
antinucleocapside

antibodies 7

Manufacturer:
Institut

Pasteur,

Paris,

Approximate cost of all media and reagents used in a year is 170 000 French
for 500 000 doses, or 0.32 French francs per dose (1986).

c:
1.

Makers and installers

BUILDINGS

of nrefabricated

buildines

- St& TRANSLCXO, France

- Ste IFFA,
2.

Builders
- Ste ESI,

France
and installers

of ventilation

France

*********

eauinment

France

francs

WJJO/Rabies/90.201

PLAN

Plan No. 1
FOR A BIOLOGICAL
PRODUCTIONS
(Housed in a hangar)

Rev.1
page 39

FACILITY

1 =
2 and
4 =
5 =
6 =
7
8
9
IO
11
12
13
14

15
16
17
18
20

Control laboratory
3 = Bacterial vaccines
Central washing plant
Media preparation
Viral vaccines (except
rabies)
= Rabies vaccine for
veterinary use
= Bottling and packaging
= Storage of finished
products and dispatch
= Workshops
= Essential common
services
= General store
= Incinerator
= Rabies animal compounds
(a) mice
(b) kennels
= Washing plant and
supplies for animal house
= Animal house (mice:
rabbits, guinea pigs)
= Administrative building
and 19 = Watchmens iodgings
= High-tension line and
transformer

11

r-----d

/!a r: I
/ !i-1I

Exit

Shaded areas = buildings under hangar

Entrance

Dimensions are given in metres

WHO 91246

WHO/Rabies/90.201
page 40

Rev.1

FIGURE 1

Placement

of

a modular

laboratory

WHO/Rabies/90.201

FIGURES 2 & 3
Example of structure

assembled

using

prefabricated

panels

Rev.1
page 41

WHO/Rabies/90.201
page 42

Rev.1

FIGURES 4 & 5
Examples

of

modular

laboratories

WHO/Rabies/90.201

Rev.1
page 43

SPECIFICATIONS OF A MODULE
(to house any type of animal house equipment)
Technical
External

dimensions

length:
breadth:
height:
Internal

snecifications

(in metres):

13.5
3
2.885

dimensions

length:
breadth:
height:

(in metres):

12.410
2.910
2.5 (at

lowest)

Panels 45 mm thick in polyester/glass

fibre/polyurethane
foam
Fittings
for the installation
of equipment
Self-supporting
floor
2 m airlock
entrance with benches, wash basin, water heater,
serving:

a)
b)

one room for standard A module

two rooms for standard B module

Day/night
Air

lighting

controlled

by timer

conditioning:
5 micron filtration
Superpressure
40% new air (standard A)
100% new air (standard B)

Maintenance

-.

a)
b)
c)

of 22"C,

50% relative

10C and 80% RH in winter

32C and 40% RH in summer
With possible
animal input

Safeguarded ventilation,
high
electrical
heating elements.
Very rapid

humidity

(RR) by:

of 1200 Kcal.
temperature,

installation.

* Example from IFFA-CREDO, France

low temperature,

refrigeration

circuit,

WHO/Rabies/90.201
page 44

Rev.1
FIGURES 6 & 7
Protection
of Personnel in sterile
areas
eg, during distribution
of media into bottles

WHO/Rabies/90.201

Rev.1
page 45

Plan No. 2
PLAN FOR A FACILITY FOR THE PRODUCTION
OF RABIES VACCINE FOR VETERINARY USE

Area 1
1
v
2
3
4
5
6
6.1
6.2
7
Al
F

=
=
=
=
=
=
=
=
=
=
=
=

Entrance hall
Dressing rooms
Room for storage in liquid nitrogen
Room for storage in freezers (- 80C)
Office
Toilets
Store
Dry ice cabinets
Crushed ice cabinets
Assembly room and sterilizers
sterilization autoclave
sterilization oven

Area 2
SI, S2 =
SMl =
=
8
=
9
CEl =
CFI =
=
10
CE2 =
=
r
=
11
SM3 =
=
12
CF2 =
Hl, H2,
=
EP

Staff entrance airlock

Airlock for media containers
Room for storage of sterile equipment
Media filtration room
Media incubation room
Media cold room
Cell culture room
Cell culture incubator
Sliding doors
Room for transfer of inactivated
virus vaccine
Transfer airlock
Room for addition of adjuvant
Cold room for bulk vaccine
H3, H4 = Laminar airflow cabinets
Pure water production

Zone 3
S3 - S4 = Staff entry airlock
SM2 = Equipment entry airlock
= Autoclave for decontamination
A2
outgoing equipment
= Viral culture room
13
CE3 = Incubator for viral culture
CF3 = Cold room for storage of
virulent harvests
= Roller doors
r
= Laminar airflow cabinet
H5

Entrance

of

Outside the building:

Production procedure: cultures in roller tubes
(system: BHK cells infected with PASTEUR PV/BHK
rabies virus)
Annual production capacity:

LTI and LT2

= Maintenance centre
and controls

400 000 to 600 000 doses.

l

Dimensions
given in metres.

WHO 91247

WHO/Rabies/90.201
page 46

Rev.1
FIGURE 8

Set of rollers

on a Bellco

unit

WHO/Rabies/90.201

Rev.1
page 47

Plan No. 3
INSPECTION UNIT

* Dimensions
given in metres
Entrance
1 =
2 =
3 =
4 =
5 and
7 =
8 =
9 =

Office
Cloakrooms and toilets
Decontamination room
Cell culture room
6 = Viral vaccine control rooms
Cold room
Biochemistry cold room
Room for physical, chemical and
biochemical analyses

IO = Room for physical, chemical and biochemical

analyses
11 = Incubation room (37)C
12 = Incubation room (30 to 35C)
13 and 14 = Bacterial vaccine control rooms
= Airlock with staff changing room
SI, s2
E = Sample library
H = Laminar airflow cabinets

WHO 91248

WHO/Rabies/90.201
page 48

Rev.1

Plan

No.

CENTRAL WASHING PLANT

13.00
Exit

* Dimensions
givenin metres

Entrance
1
2
3
4
5
6
7

=
=
=
=
=
=
=

Office
Products store room
Toilets
Changing rooms
Room for deposit of dirty equipment
Washing room
Room for preparation, assembly and
sterilization of clean equipment
8 = Store room for sterile equipment

A
B
C
D
E

=
=
=
=
=

Distillation equipment
Soaking and washing vats
Washing machines
Worktop for preparation, packaging and assembly
Autoclaves and oven

WHO 91249

WHO/Rabies/90.201

Plan

No.

Rev.1
page 49

MEDIA PREPARATION

UNIT

ti

-1

1
2
3
4
5

=
=
=
=
=

Office
Raw material store
Changing rooms and toilets
Cold room
Incubation room (37C)

6
7
Si
S2
H

=
=
=
=
=

Dimensions
given in metres

Rooms for weighing, mixing etc.

Filtration rooms
Staff entry airlock
Airlock for equipment and media to be filtered
3-p!ace laminar airflow cabinet

WHO

91250

WHO/Rabies/90.201
page 50

Rev.1
Plan No. 6
FILLING AND PACKAGING UNIT*

Shaded area is protected zone

i-l

Dimensions
given in metres

Entrance
1
2
3
5
6
7

=
=
and
=
=
=

Office
Cold room for bulk products
4 -= Filling rooms
Visual inspection room
Labelling and packaging room
Cold room for storage before dispatch

8
9
Sl
V
T

= Packing room
= Loading bay (outside)
, S2 = Airlocks with staff changing space
= Changing rooms
= Toilets
WHO 91251

WHO/Rabies/90.201

Plan

No.

Rev.1
page 51

CENTRAL ANIMAL HOUSE *

Entrance
I3
= Office
VT = Changing rooms and toilets
SAS 1 and SAS 2 = for men and women in the clean area
IAV = Washing plant
ML = Area for washing of cages
= Double-entry autoclave
A
Rl and R2 = Storage rooms for sterile equipment,
feed and bedding
Tl
= Clean corridor
T2 = Dirty corridor

Dimensions
given in metres

;;, L2 = Rabbits (breeding)

= Rabbits (keeping)
= Inspection of rabbits
L4
Cl, C2 = Guinea pigs (breeding)
c3
= Guinea pigs (keeping)
= Inspection of guinea pigs
c4
Sl, S2, S3 = Mice (breeding)
= Mice (keeping)
s4
S5, S6 = Inspection of mice
WHO 9 1252

WHO/Rabies/90.201
page 52

Rev.1
Plan

No.

PLAN OF PROTECTED ANIMAL HOUSE FOR CONTROL.OF RABIES VACCINE

FOR VETERINARY USE ON MICE AND DOGS, INCLUDING
CHALLENGE WITH VIRULENT STRAINS OF RABIES *

Area outlined in red

= protected kennel, ventilated
with negative air pressure

1
I
!
I
,I1
I
E

3
3
3

I
1
!
1
-I
I
I

E
1
Sl
SP

=
=
=
=

Wall surrounding the building

Corridor leading to kennels
Airlock, entrance for animals or equipment
Airlock, entrance for staff
(with showers and protective clothing)
1 to 20 = Individual kennels
Rl = Store room for equipment and dog food
LA = Laboratory for preparation of rabies inoculums
(wild strains) for challenges
H = BIOHAZARD vertical laminar airflow cabinet

Dimensions
given in metres

11 = Room for inoculation of dogs

A = Decontamination autoclave
2 = Corridor leading to housing for mice
Cl and C2 = Housing for mice
12 and 13 = Cubicles for inoculation of mice
R2 = Spare cages, bedding and feed for mice
52 = Decontamination airlock/exit
(for both kennels and mouse housing)
3 = Sliding doors
BU = Office

WHO 91253

WHO/Rabies/90.201

FIGURE 9

(autoclavable

stainless

Example of cage for S-10 mice

steel mesh lid,
feed tray and drinking

bottle)

Rev.1
page 53

WHO/Rabies/90.201
page 54

Rev.1

FIGURES 10 & 11
Examples

of

individual

dog cages