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    UNIVERSITI PUTRA MALAYSIA

    INFECTIOUS BOVINE RHINOTRACHEITIS VIRUS

    OF CATTLE AND BUFFALOES

    SAW PLEI SAW

    FPV 1983 4

  • INFECTIOUS BOVINE RHINOTRACHEITIS VIRUS

    OF CATTLE AND BUFFALOES

    by

    Saw Plei Saw

    A thesis submitted

    1n partial fulfilment of the requirement for the degree of Doctor of Philosophy in the

    Dept. of Veterinary Pathology and Microbiology

    Universiti Pertanian Malaysia

    June 1983

  • This thesis attached hereto, entitled "Infectious Bovine Rhinotracheitis Virus of Cattle and Buffaloes"

    prepared and submitted by Saw Plei Saw in partial

    fulfilment of the requirements for the degree of Doctor of Philosophy, is hereby accepted.

    Date:

    �� (PROFESSOR DR. ABDUL MANAP AHMAD)

    Chairman, Bo�Examinersl Dean of Graduate Studies

    Universiti Pertanian Malaysia

    t.

    External iner

    School of Veterinary Medicine

    University of California, Davis.

    DR. OMAR ABDUL RAHMAN)

    Internal Examiner

    Deputy Vice-Chancellor (Academic)

    Universiti Pertanian Malaysia

    (PROFESSOR

    Internal Examiner

    Deputy Dean

    IBRAHIM)

    Faculty of Veterinary Medicine & Animal Science Universiti Pertanian Malaysia

    8th August 1983

  • TABLE OF CONTENTS

    Chap ter

    TITLE PAGE

    APPROVAL SHEET

    TABLE OF CONTENTS

    LIST OF TABLES

    LIST OF FIGURES

    LIST OF PLATES

    ACKNOWLEDGEMENTS

    ABSTRACT

    I . INTRODUCTION

    II . REVIEW OF LITERATURE

    Ident i f icat ion of d isease and virus i s o lat ion

    C l inical manifes tations

    Morb idity and mortal i ty of IBR

    Immuni ty

    React ivation and i solat i on o f IBRV

    I solation and growth of IBRV

    IBRV purificat on and character i zation

    III . MATERIALS AND METHODS

    Serological s t udy

    I solat ion of IBRV

    Characterizat ion of IBRV

    (a) Virus purif icat ion

    (b ) Electron micros copy

    (c ) Phys ical character i s t ics

    Cd ) Cytochemical det erminat ion

    Rep licat ion of IBRV

    i i i

    Page

    1

    11

    i i i

    v

    V1

    vi i

    V111

    x

    6

    7

    1 3

    1 6

    1 7

    1 9

    24

    28

    30

    30

    33

    36

    38

    39

  • TABLE OF CONTENTS

    Chapter

    IV . RESULTS

    Sero logical f ind ings

    Reac t ivat ion of latent infect ion

    Isolat ion of IBRV

    Puri f icat ion of IBRV

    E l ectron micro s cop ic f ind ings

    Phys ical and chemical characteri s t ics

    Rep licat ion of IBRV

    v. DISCUSSION

    VI . SUMMARY

    VII . B IBLIOGRAPHY

    APPENDICES

    l.V

    Page

    4 1

    4 1

    4 2

    5 1

    56

    59

    67

    76

    87

    8 9

    1 02

  • LIST OF TABLES

    No . Page

    I . The prevalence of serum neut ral izing antibody to IBRV

    in cat tle in Malays ia

    I I . The preval ence of serum neutral i z ing ant ibody to IBRV

    in buffaloes in Malaysia

    III . Serological result shown accord ing to breed of cat t l e

    and buffaloes

    IV . Inc idence of IBR 1n cat t l e and buffaloes according

    to age group

    V . D i s tribut ion o f serum neutral i z ing ant ibody t itre

    to IBRV in catt le

    VI. D i s t r ibut ion of serum neutrali zing 'an t ibody t itre

    to IBRV in buffaloes

    VII . IBRV isolat ion from laten t ly infected catt l e

    following DM treatment

    VIII . IBRV isolat ion from latently infec ted buffaloes

    fol lowing DM treatment

    IX . The t i tre of IBRV-UPH s trains after e ther treatment

    as compared to untreated contro l

    X . The t i tre o f IBRV-UPM s t rains after treatment

    with dif ferent pH

    XI . The cell suscep t ibi l i ty range of IBRV-UPM s trains

    v

    44

    45

    46

    4 7

    4 7

    4 8

    4 9

    5 2

    6 6

    6 6

    75

  • LIST OF FIGURES

    No . Page

    1 . A map of We st Ma laysia showing locat ion of animals

    tested and found pos i t ive of IBR ant ibody

    2 . Distribut ion o f ant ibody t itre 1n cattle and

    buffaloes

    3 . Growth curve of IBRV i so lates of cat t le and

    buffaloes

    4 . A graph showing plaque size deve lopment o f IBRV 1n

    BEK cell

    V1

    43

    50

    70

    74

  • No .

    1 .

    LIST OF PLATES

    A cow showing blood t inged nasal discharge on the

    5th day of DM treatment

    2 . A cow with frothy sal ivat ion on the 5th day o f DM

    treatment

    3 . Vaginal d i s charge and hyperaemi a of the vaginal

    mucosa and pus tules format ion observed on the

    6 th day of DM treatment

    4 . Electron micrograph of negatively s t ained IBRV

    5 . Electron micrographs of ultrathin sect ion of

    BEK ce l l infected with IBRV

    6 . Electron micrograph of IBRV aggregated by IBR

    immune s erum

    7 . 16 hours IBRV infected BEK cell s ta ined with

    acri d ine orange

    8. Pho tographs showing p l aques of IBRV 1n BEK cell

    Page

    53

    54

    55

    58

    6 1 , 63

    65

    69

    73

  • ACKNOWLEDGEMENTS

    I am very much grateful to Profes sor Syed Jal aludin , the Dean of

    Faculty of Veterinary Medic ine and Animal Sc ience for provid ing me

    with fac i l i t ies required for my s tudy and also for his wise counsel

    and encouragement .

    I am profound ly grateful to Professor Abdul Latif Ibrahim for

    gIvIng me the opportunity to share his invaluab le experience in the

    f ields of virology . His ded icat ion to research works and his methodical

    approach to prob lems has much insp ired me to greater efforts . His

    pat ience In teaching and hi s cool and relaxing nature, despi te the

    many respons ib i l it ies , has provided a much rewarding exper ience to a

    few fortunate s tudents who study under his supervi s i on .

    I am also grateful to Dr . Sheikh Omar A. Rahman� Head of the

    Department of Patho logy and Microb iology for h i s interest and support

    in making th i s s tudy a succes s .

    To Dr . Abdul Az iz Saharee and Dr . Fat imah Iskandar goes my great

    appre c i a t i on for providing their exp er t is e In c l inical study and

    to Dr . Rahim Mutalib for his valuable t ips on management of laboratory

    animals .

    I woul d like to of fer my special thanks to C ik Rahmah A . Wahid ,

    Puan Rodiah Hus in and Ms . Tan Saw Eng of Viro logy unit for sharing

    their technical ski l l s and for their ever ava i lab le as s i stance and my

    VI II

  • mos t s incere gratitude to Ms Lai Chooi May, Puan Aminah Jusoh and

    Mr . Ho Oi Kuan of E lectron Microscop ic Uni t for helping me in the

    f ield of electron mi croscopy and pho tography . Above al l , I am

    much grateful to every one of them for their cordial ity which made

    my s tudy period a most enjoyab le one .

    I would l ike to expres s my heart-fel t grati tude to The

    Br it ish Counci l for award ing me the s cho lar ships and The Depar tment

    of Animal Husbandry and Veterinary Science for grant ing leave with

    ful l benef it to enable me to pursue a pos t graduate s tudy . Several

    officials and s taff help me in numerous way and I sincerely thank

    them all .

    To Ms . Low Lai Kim goes my great appreciation for her pat ience

    and promp tne s s in typ ing thi s manuscr ip t.

    Las tly , I wish to expres s my grat i tude to my parent s for their

    continuous love and encouragement and to my wife and three chi ldren

    for their love , s upport , unders tand ing and sacr i f i ce s during the

    period of s tudy .

    ix

  • ABSTRACT

    Infect ious bovine rhino trache i t i s v�rus infect ion in cat t le

    has been reported throughout the world . The s tudy of infect ious

    bovine rhinot rachei t i s in buffalo was very l imi ted . There was no

    report on infect ious bovine rhino trache i t i s in cat t le and buf falo

    in Malays ia .

    A serological s tudy on the preva lence of infect ious bovine

    rhino trache i t i s �n cat t le and buf falo in 1 0 s tates of Peninsular

    Mal ays ia showed that 52 . 5 2 per cent of the cat t le and 65 . 07 per cent

    of the buffaloes had neutralizing ant ibod ies to infec t ious bovine

    rhinotrache i t i s virus. Neutral i zing ant ibodies were detected in 532

    out of the 1 0 1 3 serum s amp les from cat t le and in 298 out of the 458

    serum samp les from buffaloes . The t itre of the serum neutral izing

    ant ibodies ranged from 1 :4 to 1 :256 .

    Four catt le and four buffaloes wi th neutral i z ing ant ibodies

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