FAO ANIMAL PRODUCTION AND HEALTH PAPER   87

COVER
Manual for the production of anthrax and blackleg vaccines
Contents


by
Dr R.P. Misra


The designations employed and the presentation of material in this publication do not imply the expression of any opinion whatsoever on the part of the Food and Agriculture Organization of the United Nations concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries.


M-27
ISBN 92-5-102920-2


All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying or otherwise, without the prior permission of the copyright owner. Applications for such permission, with a statement of the purpose and extent of the reproduction, should be addressed to the Director, Publications Division, Food and Agriculture Organization of the United Nations, Via delle Terme di Caracalla, 00100 Rome, Italy.

Preface

This manual provides a comprehensive description of standard methods for the production of vaccines against anthrax and blackleg. All steps of the procedure are taken up, from the choice of a vaccine strain to the shipping of the final product, with all relevant technical details. We have chosen to treat anthrax and blackleg vaccines in a single manual because the two diseases bear significant similarities. They are both multispecies diseases caused by spore-forming bacteria. Spores enable organisms to survive for long periods in the natural environment and particularly in soil which is a natural reservoir for both diseases.

The general methodology of vaccine production is also similar for those two organisms. In both cases the original vaccine strain can release its immunogenic properties upon repeated subcultures so that it is necessary to use a “seed system” strategy, whereby the master seed-lot is controlled for efficacy before further processing. At last both types of vaccines are evaluated for both potency and innocuity in laboratory animal models, according to standardized protocols. However, the anthrax vaccine is made from spores of an attenuated strain, whereas blackleg vaccine contains formalized cultures of a virulent strain.

We hope that this manual will assist vaccine production units of developing countries in manufacturing anthrax and blackleg vaccines of high quality standards.

FOOD AND AGRICULTURE ORGANIZATION OF THE UNITED NATIONS
Rome, © FAO 1991


Hyperlinks to non-FAO Internet sites do not imply any official endorsement of or responsibility for the opinions, ideas, data or products presented at these locations, or guarantee the validity of the information provided. The sole purpose of links to non-FAO sites is to indicate further information available on related topics.

Contents

Preface

Part I
PRODUCTION OF ANTHRAX SPORE VACCINE

GENERAL LABORATORY PROCEDURES

Environmental control

Precautions and laboratory discipline

EXPERIMENTAL ANIMALS

Guinea pigs

Sheep

Goats

MEDIA

Nutrient agar

Casein digest agar

Media for testing bacterial contamination

STRAINS OF BACILLUS ANTHRACIS

Strain for vaccine production

Strains for testing potency
Guinea pig challenge strain (17 JB)
Virulent strain

PREPARATION AND CONTROL OF VACCINE SEED-LOTS

Preparation of master seed-lot

Preparation of working seed-lot

Control of seed-lot
Tests for bacterial contamination
Safety test
Immunogenicity test

PREPARATION AND CONTROL OF
VACCINE CONCENTRATE

Preparation of vaccine concentrate
Inoculation
Harvesting
Purity test
Glycerination

Control tests on vaccine concentrate
Tests for bacterial contamination
Test for number of culturable spores
Safety test
Potency test

Dilution of vaccine concentrate

Adjuvant

FILLING AND CONTAINERS

Freeze-drying

CONTROL TESTS ON FINAL PRODUCT

Inspection of final containers

Identity test and tests for absence of bacterial contamination

Test for number of culturable spores

Determination of pH

Innocuity test

Residual moisture content for freeze-dried vaccine

Stability test

RECORDS

Records of seed-lots and cultures

Production protocol and distribution records

SAMPLES

LABELLING

DISTRIBUTION AND SHIPPING

STORAGE AND EXPIRY DATE

INSTRUCTIONS FOR USE OF VACCINE

APPENDIXES

Appendix 1
Media

Appendix 2
Procedure for detection of motility of organisms

Appendix 3
Stabilizers for freeze-drying

Appendix 4
Procedure for viable spore count

Appendix 5
Summary protocol for production and control of anthrax spore vaccine, living

Appendix 6
Certifications

REFERENCES

Part II
PRODUCTION OF BLACKLEG VACCINE

GENERAL LABORATORY PROCEDURES

Environmental control

Precautions and laboratory discipline

EXPERIMENTAL ANIMALS

Guinea pigs

Sheep

Cattle

MEDIA

Liver extract-blood agar

Liver-meat anaerobic broth

Semi-synthetic anaerobic medium

Media for testing sterility

CLOSTRIDIUM CHAUVOEI STRAIN

Cultivation

Isolation
Sampling
Isolation methods

Identification

Propagation of strain in animals

Preservation of strain
Vegetative organisms
Spores

Determination of 50-percent lethal dose (LD 50) of freeze-dried spores

Challenge dose

PREPARATION AND CONTROL OF VACCINE SEED-LOTS

Preparation of master seed-lot

Preparation of working seed-lot

Control of seed-lot
Tests for bacterial contamination
Safety test
Immunogenicity test

PREPARATION OF VACCINE

Media

Inoculum

Inoculation

Formalization

Inactivation test

Pooling

Adjuvants and concentration
Alum
Aluminium hydroxide gel

Preservation

Mixed bacterins

FILLING AND CONTAINERS

CONTROL TESTS ON FINAL PRODUCT

Inspection of final containers

Identity test

Sterility test

Determination of pH

Innocuity test

Safety test

Potency test

RECORDS

Records of seed-lots and cultures

Production protocol and distribution records

SAMPLES

LABELLING

DISTRIBUTION AND SHIPPING

STORAGE AND EXPIRY DATE

INSTRUCTIONS FOR USE OF VACCINE

APPENDIXES

Appendix 1
Media

Appendix 2
Stabilizers for freeze-drying

Appendix 3
Summary protocol for production and control of Clostridium chauvoei (blackleg) vaccine

Appendix 4
Certifications

REFERENCES