Table Of ContentHaemophilus,
Actinobacillus, and
Pasteurella
Haemophilus,
Actinobacillus, and
Pasteurella
Edited by
W. Donachie, F. A. Lainson, and
J. C. Hodgson
Moredun Research Institute
Edinburgh, Scotland
Springer Science+Business Media, LLC
L1brary of Congress Catalog1ng-1n-Pub11cat1on Data
International Conference on Haemoph11us, Act1nobac111us, and
Pasteurella <3rd : 1994: Ed1nburgh, Scotlandl
Haemoph11us, act1nobac111us, and pasteurella 1 ed1ted by H.
Donachle, F.A. La1nson, and J.C. Hodgson.
p. cm.
Includes b1b11ograph1ca1 references and Index.
ISBN 978-1-4899-0980-0 ISBN 978-1-4899-0978-7 (eBook)
DOI 10.1007/978-1-4899-0978-7
1. Heooph11us--Congresses. 2. Act1nobac111us--Congresses.
3. Pasteurella--Congresses. I. Donach1e, H. II. La1nson, F. A.
III. Hodgson, J. C. IV. T1tle.
CR82.B95I57 1994
616'.0145--dc20 95-35588
CIP
Proceedings of the Third International Conference on Haemophilus, Actinobacillus, and Pasteurella.
held July 31-August 4, 1994, in Edinburgh, Scotland, United Kingdom
ISBN 978-1-4899-0980-0
© 1995 Springer Science+Business Media New York
Originally published by Plenum Press, New York in 1995
Softcover reprint of the hardcover 1s t edition 1995
10987654321
Ali rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any
means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written
permission from the Publisher
PREFACE
The Third International Conference on Haemophilus, Actinobacillus, and Pasteurella
(HAP94) was held in July and August at the Edinburgh Conference Centre, Heriot-Watt
University, Riccarton Campus, Edinburgh, Scotland, UK. Previous conferences in 1981,
Copenhagen, and 1989, Guelph, had indicated widespread interest in this group of
pathogenic bacteria and the timing of the Edinburgh conference was prompted by the major
advances in our knowledge of the HAP group that had occurred in the five years since the
Guelph meeting. These organisms are considered as a group because of their close
relationship in an evolutionary sense and because of the similarities in the types of diseases
that they preduce. The main objectives of the meeting were to review and discuss current
knowledge and present experimental findings relating to the fundamental, applied, clinical,
and therapeutic aspects of disease research involving the HAP group of organisms.
HAP 94 was attended by 160 delegates from around the world and included many of the
foremost researchers studying HAP organisms. The conference was structured around 16
talks from invited speakers who covered key areas of HAP research including taxonomy,
mechanisms of pathogenesis, animal disease models, virulence factors, molecular biology,
immunolgy, antigen analyses, and experimental and commercial vaccine development. The
talks provided a review of the current state of research in each field and allowed each
speaker to focus on his or her personal research interests.
We are now pleased, in association with Plenum Press, to publish the conference
proceedings as a comprehensive, authoritative, and up-to-date account of many aspects of
research into the HAP group of organisms.
One area of major interest to researchers is that of plasmids and potential vectoring systems.
As a result of the great interest shown in a workshop at the conference on this topic and the
quality of the contributions presented, we have included a detailed report which we hope
will be welcomed by researchers in this field.
In addition to invited talks, the full abstracts of offered talks and posters are presented in
this book.
W.Donachie
F.A. Lainson
J.C. Hodgson
v
ACKNOWLEDGMENTS
We thank the main sponsors of the meeting, the Moredun Foundation, Lothian and
Edinburgh Enterprise Ltd., and Hoechst Animal Health Ltd., who provided the necessary
initial funding and administrative assistance essential to the setting up of the conference.
The financial help received from the United States Department of Agriculture, SmithKline
Beecham, and Solvay Duphar is gratefully acknowledged.
The cooperation of the many Moredun Research Institute staff members who were involved
in the conference and the advice and encouragement of the Director, Professor Ian Aitken,
was crucial to the success of the conference and is gratefully acknowledged. We thank
particularly the members of the Pasteurella Project Group and Mrs. Sandra McGill for their
help in organizing the conference and Mrs. Christine Curran for her advice and patience in
typing and formatting the proceedings for publication.
Lastly, we would like to record our appreciation of all the time and effort the contributors
have made to this book.
vi
CONTENTS
INVITED PAPERS
Taxonomy of the Family Pasteurellaceae Pohl 1981.. ................................................... 1
M. Bisgaard
Haemorrhagic Septicaemia (Pasteruella multocida Serotype B:2 and E:2 Infection)
in Cattle and Buffaloes ........................................................................................... 9
M.C.L. de Alwis
Vaccine Development against Pasteurella haemolytica Infections in Sheep .................. 25
W. Donachie
Viral-Bacterial Synergystic Interactions/Pathogenesis in Cattle ..................................... 39
L.A. Babiuk, M. Morsy, M. Campos, and R. Harland
Pathogenesis and Virulence of Pasteurella haemolytica in Cattle:
An Analysis of Current Knowledge and Future Approaches ................................... 51
A.W. Confer, K.D. Clinkenbeard, and G.L. Murphy
Haemophilus somnus: Antigen Analysis and Immune Responses .................................. 63
L.B. Corbeil, R.P. Gogolewski, L.R. Stephens, and T.J. Inzana
Liposaccharides and Capsules of the HAP Group Bacteria ............................................ 75
B. Fenwick
Adhesin-Receptor Interactions by Haemophilus influenzae and Other Bacteria from
the HAP Group ..................................................................................................... 89
L. van Alphen
vii
Exotoxins of Actinobacillus pleuropneumoniae ............................................................. 101
J. Frey
Receptor-Mediated Iron Acquisition from Transferrin in the Pasteurellaceae ............... 115
S.D. Kirby, J.A. Ogunnariwo, and A.B. Schryvers
Molecular Studies of Antigens in HAP Organisms ........................................................ 129
R.Y.C. Lo
Modulation of Leukocytes by Exotoxins Produced by HAP Organisms ......................... 143
C.J. Czuprynski
Anti-Idiotype Monoclonal Antibodies as LPS Internal Images and Immunologic
Protection against Endotoxin Lethality in Mice ...................................................... 153
S.K. Field, M. Pollack, and D.C. Morrison
Host Response to Infection with HAP: Implications for Vaccine Development ............. 165
P.E. Shewen
Commercial Development of Haemophilus, Actinobacillus, and Pasteurella Vaccines .. 173
S.B. Houghton
WORKSHOP SUMMARY
Genetic Manipulations of Members of the Family Pasteurellaceae ............................... 187
J. Frey and J.I. Macinnes
ABSTRACTS
Offered papers: Talks .................................................................................................... 199
Offered papers: Posters ................................................................................................. 217
Contributors .................................................................................................................. 237
Index ............................................................................................................................. 241
viii
TAXONOMY OF THE FAMILY PASTEURELIACEAE POHL 1981
M. Bisgaard
Depa11ment of Vetetinary Microbiology
The Royal Vetetinary and Agricultural University
DK-1870 Frederiksberg C
Copenhagen
Denmark
INTRODUCTION
The family Pasteurellaceae Pohl 1981 was conceived to accomodate a large group
of Gram-negative chemoorganotrophic, facultatively anaerobic, and fermentative bacteria
including the genera Pasteurella (Trevisan, 1987), Actinobacillus (Brumpt, 1910),
Haemophilus (Winslow et a!., 1917), and several other groups of organisms that exhibit
complex phenotypic and genotypic relationships with the aforementioned genera (Pohl,
1979; Pohl, 1981). Most members of the family may cause disease in mammals including
humans. birds, and/or reptiles (Bisgaard, 1993; Frederiksen, 1993).
Several methods have been used to examine the taxonomy of Pasteurellaceae.
Evidence supporting that these organisms constitute a family came from phenetic numerical
analysis (Sneath and Johnson, 1973; Broom and Sneath, 1981 ). Subsequent studies of
strains of Actinobacillus, Pasteurella and Yersinia, with some allied bacteria, showed 23
reasonable distinct groups (Sneath and Stevens, 1985), and confitmed that Actinobacillus
and Pasteurella are hard to differentiate. The existence of both V-factor dependent and
independent strains within the same species/taxon of all 3 genera has been reported for {H.]
aphrophilus (Pohl, 1979), A. pleuropnewnoniae (Pohl eta!., 1983), P. avium (Mutters et
a!., 1985a), P. multocida ssp. multocida (Krause et a!., 1987), H. parainjluenzae
(Gromkova and Koomhof, 1990), a nonclassified haemolytic Actinobacillus-like organism
(Eckart eta/., 1991), Kilian's taxon B/Bisgaard's taxon 22 (Ryll eta/., 1991), and [H.]
paragallinamm (Mouahid eta!., 1992). Studies on phenotypic traits of these organisms
thus indicate an overlapping inten-elationship between genera, and attemps to create genera
and higher taxa based upon phenotypic characterization have t-esulted in highly artificial
groupings as discussed by De Ley eta/. (1990).
THE FAMILY PASTEURELIACEAE POHL 1981
The genetic relationships of these organisms and their relationship with other
bacteria have been elucidated through DNA:DNA hybtidization (Pohl, 1979; Pohl, 1981;
Mannheim, 1983; Mutters et at., 1989), rRNA-DNA hybridization (De Ley et at., 1990),
and 16S rRNA sequencing (Dewhirst eta/., 1992; Dewhirst eta/., 1993).
Haemophilus, Actinobacillus, and Pasteurella
Edited by W. Donachie et al., Plenwn Press, New York, 1995
rRNA-DNA hybridizations showed beyond doubt that the family Pasteurellaceae
forms one cluster which is linked with both branches of the genus Alteromonas, the family
Enterobacteriaceae, the family Vibriomiceae, and the family Aeromonadaceae. All of these
clusters make up superfamily I sensu De Ley which cotTesponds to part of the gamma
subclass of the Proteohacteria. The results also showed that the family Pasteurellaceae is
as heterogeneous as the Enterobacteriaceae and Vibrionaceae and that, in addition to the
three accepted genera, there is room for at least four other new genera around each of the
following rRNA branches: "Histophilus ovis", [H.] ducreyi, [H.] aprhophi!us, and [A.]
actinomycetemcomitans. Some of these genera were quite remote from the three authentic
genera in the family.
Virtually complete 16S rRNA sequencing of representative Pasteurellaceae and
beta and gamma Proteobacteria have confiimed the location of the family in the gamma
division of the Proteohacteria (Dewhirst et al., 1992). Subsequent investigations have
shown the existence of seven major clusters, four of which contained two or more
subclusters (Dewhirst et al., 1993). The seven rRNA branches of De Ley et al. (P.
multocida NCTC l0322r, A. lignieresii NCTC 4189r, H. injluenzae NCTC 8143r, [H.]
aprhophilus NCTC 5906r, [A.] actinomycetemcomitans NCTC 9710T, H. ducreyi CIP
542r and "Histophilus ovis" HIM896-7) were recognized as subclusters 38, 4A, 1C , 1B ,
1A, 48 and 28, respectively, by Dewhirst eta!. (1993).
The general complexity of the branching within Pasteurellaceae indicates that it will
be difficult to divide the family into genera that are phenotypically coherent.
Cellular lipids and carbohydrates of the Pasteurellaceae have been reported by
Mutters eta!. (1993). The cellular fatty acid patterns proved to be uniform with minor
variations, but the separation from Neisseriaceae and Moraxella was possible. Also the
distribution of phospholipids was unifonn within the family. The lipoquinone contents were
useful for the descrimination of groups within the family, not necessarily reflecting the
degree of genomic relatedness. Finally, analysis of cellular carbohydrates showed a
common sugar pattern with all members of the family in addition to characteristic profiles
discriminating groups, often at the species level.
Polyamine patterns have been studied by Bunka et at. (1994), who detected several
different polyamine patterns among strains phylogenetically allocated the family as well as
not yet described species classified with the family, indicating that analysis of polyamine
patterns might represent useful markers for classification within the family.
As a consequence of the above mentioned studies several species previously
classified with the Pasteurellaceae have been eliminated from the family i.e. [H.]
equigenitalis, [H./ piscium, [H./ vaginalis, [P.} anatipestifer, [P.] piscicida, Pasteurella
like Bovine-lymphangitidis group and Pasteurella-like CDC group EF-4 (De Ley et a/.,
1990; Mutters eta!., 1989).
GENUS PASTEURELLA TREVISAN 1887
DNA: DNA hybridizations have shown that this genus consists of at least 11 species
some of which have not yet been named (Mutters eta!., 1989; Mutters eta!., 1985b ). The
Pasteurella sensu stricto includes P. multocida with its 3 subspecies (multocida, septica,
and gallicida), P. dagmatis, P. gallinarum, P. canis, P. stomatis, P. avium, P. volantium,
P. anatis, P. langaa and Pasteurella species A and B. Species and taxa excluded from the
genus include [P.] aerogenes, [ P.] haemolytica biovars A and T, P. multocida biovar I,
[P.} pneumotropica biovars Hey! and Jawetz, [P.] testudinis, [P.] ureae, and the SP
group.
According to De Ley et a!. (1990) strains belonging to a well defined genus are all
on the same rRNA branch. However, the P. multocida NCTC 10322T rRNA branch only
contained 3 out of 5 accepted species investigated of genus Pasteurella sensu stricto (De
2
