Thesis LJ Westerman

226
,ƵŵĂŶ /ŶƚĞƐƟŶĂů Spirochaetosis L.J. Westerman

Transcript of Thesis LJ Westerman

  • Spirochaetosis

    L.J. Westerman

    L.J. Westerm

    anH

    uman Intestinal Spirochaetosis

    2013

    Uitnodigingvoor het bijwonen van de

    openbare verdediging van mijn

    op dinsdag 10 december 2013

    om 10:30

    in het Academiegebouw van de

    Universiteit Utrecht

    Domplein 29 Utrecht

    harte uitgenodigd voor

    Kanunnikenzaal van de Faculty

    Club (achter de Senaatszaal)

    Laurens Westerman

    [email protected]

    ParanimfenMarcus Westerman

    06-46238506

    [email protected]

    Lysanne Ng-A-Foe

    [email protected]

    3605697890399

    ISBN 978-90-393-6056-9

  • Chapter 1 7

    Chapter 2 39

    Chapter 3 61

    Chapter 4 77

    Chapter 5 89

    Chapter 6 125

    Chapter 7 145

    Chapter 8 161

    Chapter 9 181

    Addendum 197

    203

    Table of Contents

  • 8

  • 9

    A brief history of the Brachyspiraceae

    .

    .

    a .

  • 10

    0.02 (~24bp)

    Figure 1:

  • 11

    .

    Figure 2:

  • 12

    Chapter 1

    4

    .

  • 13

    .

    .

    Figure 3:

  • 14

    Chapter 1

    Chapter 9

    The various Brachyspira species

    Brachyspira hyodysenteriae (hog: pig; dysentery: severe diarrhoea)

  • 15

    Tabl

    e 1:

    56

    Spec

    ies

    Indo

    leHi

    ppur

    ate

    Type

    of r

    efer

    ence

    stra

    in*

    ++

    ++

    +

    ++

    ++

    + +

    ++ +

    *

    44+ +

    ++ +

    ++

    *

    +

  • 16

    0 . 03

    ' B . i b a r a k i ' H I S 2 4 / 1 1 / 9 9

    B.innocen

    s B256

    'B.suanana' AN

    4859/03

    'B.m

    uris '

    A10

    /06

    B . a a l b o r g i 5 1 3 A

    B.a

    l vi n

    i pu

    l li

    C1

    'B .hampsoni i ' c l

    uster I NSH-16

    B.hyodysenter iae B

    78

    B. intermedia PW

    S/A

    'B.murid

    arum'

    A58/06

    B.murdoch i i 56-150

    'B.hominis' adhucomuHca09

    B. p

    i lo s

    i co l

    i P 4

    3 /6 /

    7 8

    'B.r

    aus

    ' A

    249/

    07

    ' B .hampson i i ' c lus ter I I NSH-24

    'B.corv i ' AN

    968/2/04

    ' B. p

    u ll i '

    AN

    304/

    04

    'B.ca

    nis' D

    og A2

    T .p r imi

    a ZAS - 2

    A

    0 . 0 4

    B.murd

    ochi i 5

    6-150

    B.canis Dog A2

    B. p

    u ll i

    AN3

    04/0

    4

    B. a

    l vi n

    i pu

    l li

    C1

    B.suana na AN 4859/03

    B.intermedia PWS/A

    B.ham

    pson i i c lus ter I I NSH

    -24B

    .ham

    pson

    i i

    c lus

    ter

    I N

    SH-1

    6

    B.in

    noce

    ns B

    256

    B.hyodysenter iae B78

    B.aa lborg i 513A

    B .corv i AN968/2/04B . pi l o s

    i c ol i P

    4 3 /6 / 7

    8

    T . p r i m i a Z A S - 2

    B

    Figure 4:

  • 17

    Brachyspira innocens (innocens: innocent)

    .

    Brachyspira murdochii (Murdoch: Australian university where much research into

    .

    Brachyspira pilosicoli (pilo: hairy; colon: colon)

    chapter 9

  • 18

    Chapter 1

    Brachyspira intermedia (intemediate: referring to the fact that the phenotype is between B. hyodystenteriae and B. innocens)

    ..

    In

    Brachyspira alvinipulli

  • 19

    Brachyspira aalborgi

    Brachyspira canis (canine: dog)

    p In

    Brachyspira corvi (corvus: crows)

  • 20

    Chapter 1

    44

    Brachyspira pulli

    Brachyspira muridarum and Brachyspira muris

    .

    .

    .

    (sus: pig ( Anas platyrhynchos))

  • 21

    Brachyspira hampsonii (Prof. Hampson: prominent Brachyspira

    Brachyspira ibaraki

  • 22

    Tabl

    e 2:

    Spec

    ies

    B. a

    albo

    rgi

    B. a

    lvin

    ipul

    liB.

    hyo

    dyse

    nter

    iae

    B. in

    noce

    nsB.

    inte

    rmed

    iaB.

    mur

    doch

    iiB.

    pilo

    sico

    liB

    . can

    is

    B. c

    orvi

  • 23

    Spec

    ies

    B. h

    omin

    is

    B. m

    urid

    arum

    B

    . mur

    is

    B. p

    ulli

    B. h

    amps

    onii

    B

    . ham

    pson

    ii

  • 24

    Chapter 1

    Brachyspira hominis (hominis: human)

    Zoonosis

  • 25

    Epidemiological evidence

  • 26

    Chapter 1

    .

    .

    .

  • 27

  • 28

    Chapter 1

    .

  • 29

    Figure 5:

  • 30

    Chapter 1

    Table 3:

    Country Colon biopsies Appendices Faeces*

    Concluding

  • 31

    Aim of this thesischapter 2

    chapter 4 (chapters 2 8

    In chapter 3 Chapter 5

    chapter 8. Chapter 6chapter 7

    chapter 5

  • 32

    Chapter 1

    45

    16

    41

    17

    41

    86 10

    33

    36

    11

    189

    11

    10

    1

    31

    1

    50

    49

    3

    88

    27

    12

    37

    85

    70

    82

    22

    34

    57

    31

    40

    33

    125

    63

    Int J 46

    References

  • 33

    160

    Int J 29

    Int J 47

    48

    35

    36

    28

    9

    14

    2

    24

    7

    23

    39

    53

    136

    178

    8

    67

    59

    106

    53

    130

    67

    Occurrence

    153

    15

    52

  • 34

    Chapter 1

    146

    158

    124

    137

    37

    102

    134

    46

    23

    47

    111

    23

    119

    53

    59

    146

    12

    52

    121

    99

    21

    22

    28

    53

    35

    91

    63

    32

    105

    15

    121

    36

  • 35

    41

    68

    12

    13

    61

    42

    59

    39

    2

    59

    J 22

    21

    J 113

    113

    26

    93

    146

    34

    153

    16

    158

    53

    21

    33

    64

    65

    J 31

    141

    48

    138

    143

  • 36

    Chapter 1

    7

    103

    31

    134

    179

    150

    38

    38

    224

    187

    13

    74

    134

    37

    Int J 18

    4

    45

    49

    7

    30

    49

    35

    30

    59

    101

    21

    90

    39

    41

    23

  • 37

  • 38

  • 4

    4

  • 40

    Chapter 2

    Abstract

    Methods

    Results

    p

    Conclusions

  • 41

    4

    Jensen

  • 42

    Chapter 2

    .

    Aims of this study

  • 43

    MethodsEthics statement

  • 44

    Chapter 2

    Processing of FFPE biopsy samples

  • 45

  • 46

    Table 1:

    Primer set 53 Total length C Time Purpose

    PCR volume

    PCR consumables

    PCR protocol

  • 47

    m

    Sequencing

    Figure 1:

    StepA (212bp)60 271

    StepB (253bp)203 445

    StepC (258bp)366 623

    Bspp (82bp)135 216

    5 31000 200 300 400 500 600 700

    AmpliconPrimer

  • 48

    Chapter 2

  • 49

    Results

    A

    PCR results

    p

    p p

  • 50

    -2

    0

    2

    4

    6

    8

    10

    12

    66 68 70 72 74 76 78 80 82 84 86 88 90 92 94 96

    B. aalborgi)

    B. pilosicoli)

    Figure 2:

    Table 2

    Sequence result

  • 51

    B. pilosicoli

    B. burgdorferi

    B. murdochii B. innocens

    B. hyodysenteriae B. intermedia

    B. alvinipulli

    B. aalborgi

    0.01

    B. pilosicoli

    Brachyspira

    B. hominis

    Brachyspira aalborgi

    Figure 3:

  • 52

    Chapter 2

    (p

    Table 3:

    p

    # + +

  • 53

    Discussion

    .

  • 54

    Chapter 2

    m

  • 55

  • 56

    Chapter 2

  • 57

    48

    35

    160

    2

    Int J 46

    40

    67

    63

    106

    62

    41

    16

    12

    53

    39

    References

    23

    53

    42

    16

    31

    49

    7

    41

    115

    12

    2

    39

    59

    116

  • 58

    Chapter 2

    197

    25 Suppl 3

    37

    27

    12

    132

    83

    95

    32

    23

    40

    67

    136

    178

    35

  • 59

    Supplementary Table S1:

  • 60

  • 62

    Chapter 3

    Abstract

    Methods

    Results

    Conclusion

  • 63

    Appendiceal Spirochaetosis in Children

    .

    .

  • 64

    Chapter 3

    .

  • 65

    Appendiceal Spirochaetosis in Children

    Materials and methodsEthics statement

    Sample processing

  • 66

    Chapter 3

  • 67

    Table 1:

    53 Amplicon

    PCR volume

    PCR protocol

  • 68

    Chapter 3

    Histopathology

  • 69

    Appendiceal Spirochaetosis in Children

    Results

    Table 2:

    Immunohistochemistry

    PCR result + ++ +++

    p

    (p

  • 70

    Chapter 3

    Discussion

    Table 3:

    Total appendices Control group Other diagnoses

    4

  • 71

    Appendiceal Spirochaetosis in Children

    (p

  • 72

    Chapter 3

  • 73

    Appendiceal Spirochaetosis in Children

    24

    17

    12

    27

    88

    90

    23

    8

    3

    16

    7

    160

    67

    35

    2

    142

    2

    59

    39

    53

    39

    23

    154

    52

    J 38

    53

    4

    31

    Am J 132

    55

    25 Suppl 3

    51

    7

    References

  • 74

    Chapter 3

    37

    33

    65

  • 75

  • 76

  • 44

  • 78

    Chapter 4

    Abstract

    (p

  • 79

    As a

  • 80

    Chapter 4

    Materials and methods

    Table 1: a

    a

  • 81

    Results

    p

    p

  • 82

    Chapter 4

    Discussion

    p

  • 83

  • 84

    Chapter 4

  • 85

    16

    3

    7

    53

    23

    59

    2

    J 22

    45

    82

    4

    53

    J 38

    42

    16

    31

    7

    48

    85

    101

    21

    21

    39

    70

    82

    22

    53

    132

    31

    References

  • 86

    Supplementary table S1:

    Sample ID

    BacteriaParasites

    S. entericaC. jejuni

    S. dysenteriaeSTEC/EPEC

    G. lam

    bliaD. fragilis

    C. parvum

    +++

    *+

    +*

    *+

    *+

    *

    +*

    *

    +#

    +++

  • 87

  • 88

  • A

    4

    4

  • 90

    Chapter 5

    Abstract

  • 91

    Brachyspira species

    .

    (

  • 92

    Chapter 5

    . In

    (alp

    .

  • 93

    Material and methodsMLST design

    Table 1:

    Criterion Database

  • 94

    Chapter 5

    atpG

  • 95

    atpG

  • 96

    Table 2:

    Locus TargetAmplicon (bp)

    Primers**Total Useful Min

    atpG

    PCR volume

    PCR consumables

    PCR protocol

  • 0.02 (~24bp)

    Colour-plate A:

  • Colour-plate B:

  • Colour-plate C:

  • -2

    0

    2

    4

    6

    8

    10

    12

    65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96

    513A (B. aalborgi)

    WesB (B. pilosicoli)

    Colour-plate D:

  • -2

    0

    2

    4

    6

    8

    10

    12

    65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96

    513A (B. aalborgi)

    WesB (B. pilosicoli)

    Colour-plate E:

  • B. aalborgi

    B. pilosicoli

    B. hominis

    B. hyodysenteriae

    B. murdochii

    B. alvinipulli

    B. innocens

    B. intermedia

    Colour-plate F:

  • B. innocens

    B. intermedia

    B. aalborgi

    B. pilosicoli

    B. hominis

    B. hyodysenteriae

    B. murdochii

    B. alvinipulli

    B. innocens

    B. intermedia

    Colour-plate G:

  • Colour-plate H:

  • Colour-plate I:

  • Colour-plate J:

    0.05

    WW25

    200900828

    H4-2

    200909930

    Q95.3281.0

    WF1

    201124725

    Q97.2224.3.1

    201221374

    200513182a

    Q95.0635.6

    SC1

    Q98.0026.12

    VTN021f

    513A

    HRM_4BHRM_2B

    Q95.2078.2

    Q97.2110.4.11

    3295.90BW015/C138

    Q1588.5

    EM1

    OF_11

    199800660

    Q97.000.6.8

    N1

    QF1

    HRM7A

    7072i

    201102362

    Q98.0033.72

    201204927

    P43/6/78

    Q98.0072.08

    200831665

    201009156

    K1

    B256

    B-78

    Q98.0037.6

    T.primitia_ZAS-2

    HRM7

    201101966

    T1

    9803.1

    PWS/A

    f199600014

    201105386

    KP5

    201013692

    H21

    200410620a

    M1

    V1_H141

    L72200300944

    WesB

    201125230

    WIA8

    C1

    D9201243

    V1_H11

    201009139

    201200678VTN022f

    RV1

    Q1404.6A

    200410620b

    H38-2

    H60-2

    Q91.1223.3

    200805431

    200517505

    Q98.0028.3

    J_F2

    200709033

    WA1

    95/1000

    200232081

    200513182b

    Q94.0354.0.6

    56-150

    Q98.0072.31

    Human

    Pig

    Chicken

    Dog

  • 0.05

    9803.1

    T1

    HRM7A

    D9201243

    Q91.1223.3

    HRM_4B200909930

    HRM_2B

    WW25

    Q95.3281.0

    SC1

    Q98.0072.31

    200805431

    201009156

    W015/C138

    JF2

    B-78

    Q94.0354.0.6

    201200678

    N1

    OF_11

    200709033

    M1

    C1

    V1_H141

    200300944

    201221374

    WA1

    56-150

    200513182a

    200517505

    H21

    7072i

    201013692

    Q98.0028.3

    201101966

    VTN022f

    Q95.2078.2

    201125230

    WesB

    95/1000

    Q97.2224.3.1

    Q1404.6A

    Q97.2110.4.11Q95.0635.6

    VTN021f

    WIA8

    200513182b

    200831665

    201009139

    201102362

    P43/6/78

    QF1

    200410620b

    Q97.000.6.8

    513A

    201105386

    Q98.0026.12

    200900828

    200232081

    201204927

    WF1

    Q1588.5

    3295.90B

    200410620a

    PWS/A

    Q98.0072.08

    HRM7

    EM1

    H38-2

    K1

    KP5

    H60-2

    T.primitia_ZAS-2

    201124725

    B256

    RV1

    V1_H11

    H4-2

    A

    B

    C

    Human

    Pig

    Chicken

    Dog

    Colour-plate K:

  • Human PigChicken Dog

    Colour-plate L:

    The Netherlands

    Australia

    Italy

    Papua New Guinea

    United Kingdom

    United States of America

  • The Netherlands

    Australia

    Italy

    Papua New Guinea

    United Kingdom

    United States of AmericaColour-plate M:

  • OverijsselSIR biopsies: 0.94 (0.93 - 0.95)

    SIR HIS: 0.96 (0.72 - 1.24)Pigs 483.2/km2

    Poultry 2,943.3/km2

    HIV-centres 2/30

    GelderlandSIR biopsies: 1.08 (1.08 - 1.09)

    SIR HIS: 0.48 (0.36 - 0.64)Pigs 442.9/km2

    Poultry 3,439.3/km2

    HIV-centres 2/30

    DrentheSIR biopsies: 0.92 (0.90 - 0.93)

    SIR HIS: 0.54 (0.29 - 0.90)Pigs 107,5/km2

    Poultry 2,363.8/km2

    HIV-centres 0/30

    FrieslandSIR biopsies: 1.04 (1.02 - 1.05)

    SIR HIS: 0.21 (0.08 - 0.42)Pigs 31.8/km2

    Poultry 2,187.2/km2

    HIV-centres 1/30

    GroningenSIR biopsies: 0.89 (0.88 - 0.90)

    SIR HIS: 0.29 (0.13 - 0.55)Pigs 63.7/km2

    Poultry 2.376.6/km2

    HIV-centres 1/30

    FlevolandSIR biopsies: 1.20 (1.18 - 1.22)

    SIR HIS: 2.24 (1.62 - 3.02)Pigs 40.8/km2

    Poultry 1,461.9/km2

    HIV-centres 2/30

    UtrechtSIR biopsies: 0.97 (0.96 - 0.98)

    SIR HIS: 0.93 (0.71 - 1.20)Pigs 219.1/km2

    Poultry 1,227.9/km2

    HIV-centres 2/30

    Noord-HollandSIR biopsies: 1.01 (1.01 - 1.02)

    SIR HIS: 1.84 (1.63 - 2.08)Pigs 9.7/km2

    Poultry 493.4/km2

    HIV-centres 9/30

    Zuid-HollandSIR biopsies: 0.99 (0.98 - 0.99)

    SIR HIS: 0.46 (0.37 - 0.57)Pigs 56.5/km2

    Poultry 390.0/km2

    HIV-centres 7/30

    Noord-BrabantSIR biopsies: 0.82 (0.81 - 0.82)

    SIR HIS: 0.58 (0.46 - 0.73)Pigs 1,078.6/km2

    Poultry 5,330.5/km2

    HIV-centres 2/30

    LimburgSIR biopsies: 1.27 (1.26 - 1.28)

    SIR HIS: 2.92 (2.51 - 3.38)Pigs 790.6/km2

    Poultry 6,880.0/km2

    HIV-centres 1/30

    ZeelandSIR biopsies: 0.86 (0.85 - 0.88)

    SIR HIS: 1.03 (0.64 - 1.58)Pigs 43.6/km2

    Poultry 1,015.2/km2

    HIV-centres 1/30

    Colour-plate N:

  • OverijsselSIR biopsies: 0.94 (0.93 - 0.95)

    SIR HIS: 0.96 (0.72 - 1.24)Pigs 483.2/km2

    Poultry 2,943.3/km2

    HIV-centres 2/30

    GelderlandSIR biopsies: 1.08 (1.08 - 1.09)

    SIR HIS: 0.48 (0.36 - 0.64)Pigs 442.9/km2

    Poultry 3,439.3/km2

    HIV-centres 2/30

    DrentheSIR biopsies: 0.92 (0.90 - 0.93)

    SIR HIS: 0.54 (0.29 - 0.90)Pigs 107,5/km2

    Poultry 2,363.8/km2

    HIV-centres 0/30

    FrieslandSIR biopsies: 1.04 (1.02 - 1.05)

    SIR HIS: 0.21 (0.08 - 0.42)Pigs 31.8/km2

    Poultry 2,187.2/km2

    HIV-centres 1/30

    GroningenSIR biopsies: 0.89 (0.88 - 0.90)

    SIR HIS: 0.29 (0.13 - 0.55)Pigs 63.7/km2

    Poultry 2.376.6/km2

    HIV-centres 1/30

    FlevolandSIR biopsies: 1.20 (1.18 - 1.22)

    SIR HIS: 2.24 (1.62 - 3.02)Pigs 40.8/km2

    Poultry 1,461.9/km2

    HIV-centres 2/30

    UtrechtSIR biopsies: 0.97 (0.96 - 0.98)

    SIR HIS: 0.93 (0.71 - 1.20)Pigs 219.1/km2

    Poultry 1,227.9/km2

    HIV-centres 2/30

    Noord-HollandSIR biopsies: 1.01 (1.01 - 1.02)

    SIR HIS: 1.84 (1.63 - 2.08)Pigs 9.7/km2

    Poultry 493.4/km2

    HIV-centres 9/30

    Zuid-HollandSIR biopsies: 0.99 (0.98 - 0.99)

    SIR HIS: 0.46 (0.37 - 0.57)Pigs 56.5/km2

    Poultry 390.0/km2

    HIV-centres 7/30

    Noord-BrabantSIR biopsies: 0.82 (0.81 - 0.82)

    SIR HIS: 0.58 (0.46 - 0.73)Pigs 1,078.6/km2

    Poultry 5,330.5/km2

    HIV-centres 2/30

    LimburgSIR biopsies: 1.27 (1.26 - 1.28)

    SIR HIS: 2.92 (2.51 - 3.38)Pigs 790.6/km2

    Poultry 6,880.0/km2

    HIV-centres 1/30

    ZeelandSIR biopsies: 0.86 (0.85 - 0.88)

    SIR HIS: 1.03 (0.64 - 1.58)Pigs 43.6/km2

    Poultry 1,015.2/km2

    HIV-centres 1/30

    1 Appendix2 Coecum3 Ascending colon4 Transversing colon5 Descending colon6 Sigmoid7 Rectum

    12

    3

    4

    5

    67

    A

    12

    3

    4

    5

    67

    B

    12

    3

    4

    5

    67

    C

    12

    3

    4

    5

    67

    D

    Colour-plate O:

  • Colour-plate P:

    A

    B

  • 97

    ResultsSelected MSLT genes

    ( atpG

    atpG

  • 98

    0 . 05

    201122

    867_B.

    aalborg

    i

    201022967g_B.aalborgi

    95 /1000_GenBank_B.p i los ico l i

    201024300_B.aalborgi

    ilocisolip.B

    _669101102

    201105351b_B.aalborgi

    B256_B.innocens

    200704900_B.aalborgi

    200216407_B.aalborgi

    ilocis

    olip.B

    _931

    9001

    02200410620a_B .p i l os i co l i

    201129345_B.aalborgi

    PWS/A_B

    .intermedia

    sinimoh.B_b295300102

    200920244_B.aalborgi

    ilocis

    olip.

    B_44

    9003

    002

    200917097_B.aalborgi

    201050465_B.aalborgi

    20111037

    4b_B.aalb

    orgi

    ilocisolip.B_

    032521102

    200232081_B .p i l o s i co l i

    igroblaa.B_e153501102

    200902757_B.

    aalborgi

    201003592a_B

    .hom

    inis

    200913478_

    B.aalborgi

    201022967d_B.aalborgi

    200430368_B.aalborgi

    2011

    1160

    3_B.aalbo

    rgi

    iloc

    isoli

    p.B_

    6519

    0010

    2

    ilocisolip

    .B_26320

    1102

    201104608_B.aalbo

    rgi

    200813138f_B.aa lborg i

    200813138a_B.aalborgi

    200813138g_B.aalborgi

    201022967b_B.aalborgi

    201122669_B.aalborgi

    201120

    460_B.

    aalbor

    gi

    iloc

    isoli

    p.B_

    7294

    0210

    2

    200805639_B.aalborgi

    200813138e_B.aalborgi

    200813340_B.a

    alborg

    i

    200922746_B.aalborgi

    200925995_B.hominis

    201003

    249_B.

    aalborgi

    201021258_B.hominis

    200414761_B.aalborgi

    200811646_B.aalborgi

    201024348_B.hominis

    200608126*_B.hom

    inis

    sinimoh.

    B_343806002

    200813138d_B.aalborgi

    201013692_B .p i l o s i co l i

    2011

    2389

    8_B.aalbo

    rgi

    200703075_B.a

    alborg

    i

    201108603_B.hom

    inis

    201024610_B.aalborgi

    igro

    blaa

    .B_

    c153

    5011

    02 igroblaa.B_d153501102

    20111037

    4d_B.aalb

    orgi

    200709653_B.aalborgi

    200813138c_B.aalborgi

    201003592c_B

    .hom

    inis

    201022967h_B.aalborgi

    ilocis

    olip.B

    _828

    0090

    02

    200501138_B.a

    alborg

    i

    sini

    moh.

    B_70

    0611

    102

    201104140_B.hominis

    200918072_B.aalborgi

    201150177_B.aalborgi

    iloc

    isoli

    p.B_

    4731

    2210

    2

    201024466_B.aalborgi

    200602985_B.aalborgi201

    110374

    a_B.aal

    borgi

    iloc

    isoli

    p.B_

    5274

    2110

    2

    201150471_B.aalborgi

    200113332_B.aalborgi

    ilocisolip

    .B_6835

    01102

    201014350_B.aalborgi

    201040076_B.aalborgi

    201110374g

    _B.aalborgi

    ilocis

    olip.B

    _03990

    9002

    201129

    144_B.

    aalbor

    gi

    200901135_

    B.aalborgi

    iloc

    isoli

    p.B_

    0660

    0899

    1

    201104406_B.aalborgi

    201015580_B.aalborgi

    56-150_B.murdochi i

    C1 _ B

    . a l v i n i p u l l i

    200808214_B.a

    alborg

    i

    200914805_B.aalborgi

    201018

    213_B.aa

    lborgi

    B-78_B

    .hyodysenter iae

    2011

    1430

    3_B.aalbo

    rgi

    T. p r im

    i t i a_ZAS-2

    sini

    moh.

    B_30

    0706

    00220

    0803874_B.hominis

    200513182b_B .p i l os i co l i

    200923973_B.a

    alborgi

    201004

    769_B.aa

    lborgi

    201103

    059_B.

    aalbor

    gi

    sini

    moh.

    B_56

    6207

    002

    200624827_B.aalborg

    i

    20111193

    3_B.aalbo

    rgi

    P 4 3 / 6 / 7 8 _B . p i l o s i c o l i

    201104149_B.aalborgi

    iloci

    soli

    p.B_

    5057

    1500

    2

    ilocisolip.B_5661380

    02201023408_B

    .aalborgi

    201119045_B.aalborgi

    2011

    0535

    1g_B

    .aalbo

    rgi

    2 00513182a_B .p i l os i co l i

    sinimoh.

    B_207415002

    201202009_B.aalborgi

    200702703_B.aa

    lborgi

    201113577_B.aalborgi

    200813138h_B.aa

    lborgi

    200901

    634_B.a

    alborgi

    200101917_B.aalborgi201022967i_B

    .aa lborg i

    201050211_B.aalborgi

    igro

    blaa

    .B_

    a153

    5011

    02

    iloc

    isoli

    p.B_

    3309

    0700

    2

    201109833_B.aalborgi

    WesB_B .p i l o s i co l i

    201124639_B.aalborgi

    200900639_B.aalborgi

    200707443_B.aalborgi

    201003592d_B

    .hom

    inis

    201022967c_B.aalborgi

    200716125_B.aalborgi

    201022967f_B.aa lborg i

    igroblaa.B_f153501102

    20081681

    7_B.aalb

    orgi

    200904

    021_B.aa

    lborgi

    201108681_B.aalbo

    rgi

    201022967e_B.aalborgi

    ilocisolip.B_502306991

    200813138b_B.aalborgi

    513A_B.aa

    lborgi

    iloc

    isoli

    p.B_

    8760

    0210

    2

    iloc

    isoli

    p.B_

    1345

    0800

    2

    201003592e_B

    .hom

    inis

    201001908*_B.aalborg i

    200410620b_B .p i l os i co l i

    sinimoh.

    B_115509002

    200917337_B.aalborgi

    201007259_B.aalborgi

    WA1_G

    enBank_B.hyodysenteriae

    201102752_B.aalborgi

    Figure 1:19

    200813

    138a_B

    .aalborg

    i

    200918072_

    B.aalborgi

    200813138c_B.aalborgi

    200923973_B.aalborgi

    201129345_B.aalborgi

    20081313

    8d_B.aalb

    orgi

    201022967d

    _B.aalborgi

    201150471_B.aalborgi

    200813138f_B.aa lborg i

    200805639_B.aalborgi

    200917097_B.aalborgi

    200608126*_B.hominis

    201022967f_B.aa lb

    org i

    201022967g_B

    .aalborgi

    200430368_B.aalborgi

    20081313

    8g_B.aalb

    orgi

    201104140_B.hominis

    201022967c_B.aalborgi

    200900639_B.aalborgi

  • 99

    200704900_B.aalborgi

    20111037

    4b_B.aalb

    orgi

    200902757_B.

    aalborgi

    201003

    249_B.

    aalborgi

    20111037

    4d_B.aalb

    orgi201

    110374

    a_B.aal

    borgi

    201110374g

    _B.aalborgi

    200624827_B.aalborg

    i2007027

    03_B.aalborgi

    200917337_B.aalborgi

    0 . 06

    200216407_B.aalborg

    i

    2007

    0744

    3_B.aalbo

    rgi

    2 00410620a_B .p i l os i co l i

    201105351d_B

    .aalborgi

    200813

    138a_B

    .aalborg

    i

    200513182b_B .p i l os i co l i

    200813340_B.a

    alborg

    i

    201120

    460_B.

    aalborgi

    iloc

    isoli

    p.B_

    7294

    0210

    2

    201003592b_B.hominis

    201111933_B.a

    alborg

    i

    WA1_GenBank_B.hyodysenteriae

    201024466_B.aalborgi

    B256_B

    .innocens

    201109833_B.aalborgi

    200918072_

    B.aalborgi

    201004769_B.aalborgi

    200414761_B.aalborgi

    200813138c_B.aalborgi

    iloc

    isoli

    p.B_

    5057

    1500

    2

    201111603_B.aalborgi

    201007259_B.aalborgi

    201003249_B.aalborgi

    200923973_B.aalborgi

    200702665_B.hominis

    200905

    511_B.

    hominis

    PWS/A_B

    .intermedia

    200913478_B.aalborgi

    201022967h_B

    .aalborgi

    201129345_B.aalborgi

    2007

    0965

    3_B.aalbo

    rgi

    igroblaa.B_*809100102

    2011

    0868

    1_B.aalbo

    rgi

    201104608_B.a

    alborg

    i

    igro

    blaa

    .B_

    5703

    0700

    2

    200914805_B.aalborgi

    ilocisolip.B_066008991

    200716125_B.aalborgi

    201113577_B.aalborgi

    20111037

    4a_B.aalb

    orgi

    ilocisol

    ip.B_1

    345080

    02

    201050211_B.aalborgi

    200901

    634_B.aa

    lborgi

    C1_B

    . a l v i n i p u l l i

    201024300_B.aalborgi

    201014350_B.aalborgi

    200902757_B.aa

    lborgi

    201110374d

    _B.aalborgi

    20081313

    8d_B.aalb

    orgi

    201022967d

    _B.aalborgi

    200113332_B.aalborgi

    200808214_B.aalborgi

    95 /1000_GenBank_B.p i los ico l i

    ilocis

    olip.B

    _566

    1380

    02

    201202009_B.aalborgi

    201003592c_B.hominis

    201123898_B.aalborgi

    200514702_B.hominis

    200813138h_B.aalborgi

    201122867_B.aalborgi

    201003592d_B.hominis

    2010

    2340

    8_B.aalbo

    rgi

    iloc

    isoli

    p.B_

    6835

    0110

    2

    ilocis

    olip.B

    _33090

    7002

    201022967e_B.aalborgi

    201150471_B.aalborgiigroblaa.

    B_647229002

    200813138f_B.aa lborg i

    201103

    059_B.

    aalbor

    gi

    200920

    244_B.aa

    lborgi

    P 4 3 / 6 / 7 8 _B . p i l o s i c o l i200813138e_B.aalborgi

    200805639_B.aalborgi

    B-78_B.hyodysenter iae

    2010

    2461

    0_B.aalbo

    rgi

    201040076_B.aalborgi

    ilocis

    olip.B

    _263

    2011

    02

    201021258_B.hominis

    201003592e_B.ho

    minis

    201105351a_B.aalborgi

    20111037

    4b_B.aalb

    orgi

    ilocisol

    ip.B_87

    600210

    2

    201114303_B.aalborgi

    200917337_B.a

    alborg

    i

    201102

    752_B.

    aalbor

    gi

    200816817_B.aalborgi

    ilocisolip

    .B_6519

    00102200410620b_B .p i l os i co l i

    iloc

    isoli

    p.B_

    6691

    0110

    2

    iloc

    isoli

    p.B_

    5023

    0699

    1

    200917097_B.aalborgi

    200608343_B.hominis

    201104149_B.aalborgi

    200513182a_B .p i l os i co l i

    ilocisolip.B_8280090

    02

    200608126*_B.hominis

    ilocisolip.B_296

    310102

    201150177_B.aalborgi

    201022967f_B.aa lb

    org i

    201022967g_B

    .aalborgi

    201050465_B.aalborgi

    iloc

    isoli

    p.B_

    0399

    0900

    2

    201015580_B.aalborgi

    201104406_B.aalborgi

    200430368_B.aalborgi

    iloc

    isoli

    p.B_

    4490

    0300

    2

    201105351f_B

    .aa lborgi

    200624827_B.aalborgi

    201105351g_B

    .aalborgi

    201122

    669_B.aa

    lborgi

    igroblaa.B_009407002

    201018213_B.aalborgi

    200925995_B.hominis

    201009139_B .p i l o s i co l i

    2011

    0535

    1c_B

    .aalbo

    rgi

    20081313

    8g_B.aalb

    orgi

    201022967i_B.aa lborg i

    201003592a_B.hominis

    200602

    985_B.

    aalborg

    i

    sinimoh.

    B_843420102

    2009

    0402

    1_B.aalbo

    rgi

    200607003_B.hominis

    201116007_B.hominis

    513A_B.aa lborg i

    201104140_B.hominis

    201110374g_B

    .aalborgi

    2011

    0535

    1b_B

    .aalbo

    rgi

    201119045_B.aalborgi

    WesB

    _B.p i l o s i co l i

    iloc

    isoli

    p.B_

    5274

    2110

    2

    igro

    blaa

    .B_

    3072

    0700

    2

    201022967c_B.aalborgi

    iloci

    soli

    p.B_

    4731

    2210

    2

    201105351e_B

    .aalborgi

    igro

    blaa

    .B_

    8311

    0500

    2

    200101917_B.aalborgi

    sini

    moh.

    B_47

    8308

    002

    201108603_B.hominis

    200900639_B.aalborgi

    ilocisolip.B_

    032521102

    ilocis

    olip.

    B_18

    0232

    002

    201124

    639_B.a

    alborgi

    201022967b_B.aalborgi

    igro

    blaa

    .B_

    5311

    0900

    2

    200813138b_B.aalborgi

    igroblaa.B_441921102

    56-150_B.m

    urdochi i

    200811646_B.aalborgi

    T.pr im

    i t i a_ZAS-2

    Figure 2:

  • 100

    0 . 06

    200709653_B.a

    alborg

    i

    iloc

    isoli

    p.B_

    0399

    0900

    2

    WesB_B .p i l o s i co l i

    201114303_B.aalborgi

    ilocisolip.B_566138002

    200702665_B.hominis

    200813138b_B.aalborgi

    201150471_B.aalborgi

    200624827_B.aalborgi

    201021258_B.hominis

    igro

    blaa

    .B_

    9414

    0110

    2

    201108681_B.a

    alborg

    i

    201122

    867_B.aa

    lborgi

    200816

    817_B.

    aalbor

    gi

    iloc

    isoli

    p.B_

    6835

    0110

    2

    200902757_B.aalborgi

    igroblaa.B_936421102

    200914

    805_B.aa

    lborgi

    200602985_B.aalborgi

    200514702_B.hominis

    2009

    0402

    1_B.aalbo

    rgi

    200608343_B.hominis

    iloc

    isoli

    p.B_

    0325

    2110

    2200811646_B.aalborgi

    200917337_B.aalborgi

    C 1 _ B . a l v i n i p u l l i

    iloc

    isoli

    p.B_

    a026

    0140

    02

    201003592c_B.hominis 20

    0805639_B.aalborgi

    201015580_B.aalborgi

    201003592d_B.hominis

    B256_B.innocens

    201022967e_B.aalborgi

    200803874_B.hominis

    200920244_B.aalborgi

    201104608_B.a

    alborg

    i

    200925995_B.hominis

    T.pr imi ti a_Z

    AS-2

    201123898_B.aalborgi

    200513182a_B .p i l os i co l i

    WA1_GenBank_B.hyodysenteriae

    200917097_B.aalborgi

    201129144_B.aalborgi

    ilocis

    olip.

    B_50

    5715

    002

    iloc

    isoli

    p.B_

    1345

    0800

    2

    201110374d_B

    .aalborgi

    200900639_B.

    aalborgi

    200813

    138g_B

    .aalborg

    i

    ilocis

    olip.B

    _18023

    2002

    201116007_B.hominis

    iloc

    isoli

    p.B_

    8760

    0210

    2

    ilocis

    olip.B_

    473122

    102

    PWS/A_B.intermedia

    201007259_B.aalborgi

    200901135_B.aalborgi i

    locis

    olip.

    B_44

    9003

    002

    201022967d_B.aalborgi

    201111603_B.aalborgi 2

    0115

    0177

    _B.aalbo

    rgi

    95 /1000_GenB

    ank_B.p i los ico l i

    201110374b_B

    .aalborgi

    201018213_B.aalbo

    rgi

    ilocisolip

    .B_6691

    01102

    201050

    465_B.

    aalbor

    gi

    iloc

    isoli

    p.B_

    2963

    1010

    2

    200813

    138a_B

    .aalbor

    gi

    20091807

    2_B.aalb

    orgi

    201022967c_B.aalborgi

    200702703_B.aalborgi

    200414761_B.aalborgi

    20110440

    6_B.aalbo

    rgi

    iloc

    isoli

    p.B_

    b026

    0140

    02

    200813138d_B.aalborgi

    200501138_B.aa

    lborgi

    201119045_B.aalborgi

    201111933_B.aalborgi

    B-78_B.hyodysenter iae

    P43 / 6 / 7 8 _B . p i l o s i c o l i

    200913478_

    B.aalborgi

    200922746_B.aalborgi

    200113332_

    B.aalborgi

    200813138e_B.aalborgi

    iloc

    isoli

    p.B_

    6519

    0010

    2

    200716

    125_B.

    aalborgi

    513A_B.aa lborg i

    ilocisolip.B_263201102

    201003249_B.aalborgi

    201105351b_B.aalborgi

    iloc

    isoli

    p.B_

    8280

    0900

    2

    201105351c_B.aalborgi

    201105351d_B.aalborgi

    200707443_B.aalborgi

    20070490

    0_B.aalb

    orgi

    200513182b_B .p i l os i co l i

    ilocisolip.B_330907002

    201024610_B.aalborgi

    201050211_B.aalborgi

    20081313

    8c_B.aalb

    orgi

    56-150_B.murdochi i

    200905511_B.hominis

    201103059_B.aalborgi

    iloc

    isoli

    p.B_

    9319

    0010

    2

    201104140_B.hominis

    201202009_B.aalborgi

    201122669_B.aalborgisi

    nimo

    h.B_

    8434

    2010

    220

    0923

    973_

    B.aalbo

    rgi

    igroblaa.B_053410102

    200808214_B.aalborgi

    iloc

    isoli

    p.B_

    7294

    0210

    2

    201024300_B.a

    alborg

    i

    201003592b_B.hominis

    iloc

    isoli

    p.B_

    5274

    2110

    2

    Figure 3:

    201113577_B.aalborgi

    201129345_B.aalborgi

    200430368_B.aalborgi200716125_B.aalborgi

    200923973_B.aalborgi

    20111600

    7_B.homi

    nis

    200914805_B.aalborgi

    201003249_B.aalborgi

    20110440

    6_B.aalb

    orgi20115

    0177_B.aal

    borgi

    201015580_B.aalbo

    rgi

    201122669_B.aalborgi

    200816817_B.aalborgi

    201108681_B.aalborgi

    201150471_

    B.aalborgi

    201102752_B.aalborgi

    201050

    465_B.a

    alborgi

    200917337_B.aalborgi

    201202009_B.a

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  • 101

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    20081313

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    Figure 4:

  • 102

    0 . 06

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    igroblaa.B_646118002

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    ilocisolip.B_931900102

    T . p r im i t i a_ZAS-2

    Figure 5:

  • 103

    T .p r im i t i a_ZAS-2

    atpG

    Table 3:

    Locus MLST Length (bp)variable sites

    AATLength

    variable sites

    atpG

    **

  • 104

    Figure 6:

    B. aalborgi

    B. hominis

    B. pilosicoli

    B. hyodysenteriae

    B. murdochii

    B. alvinipulli B. intermedia

    B. innocens

  • 105

    B. intermedia

    B. aalborgi

    B. hominis

    B. pilosicoli

    B. hyodysenteriae

    B. murdochii

    B. alvinipulli B. intermedia

    B. innocens

    Figure 7:

  • 106

    Chapter 5

  • 107

    Figure 8:

  • 108

    Chapter 5

    Discussion

  • 109

    As

    atpG

    .

  • 110

    Chapter 5

    .

  • 111

  • 112

    Chapter 5

    32

    4

    36

    36

    3

    77

    153

    43

    14

    43

    62

    References

    2

    154

    52

    39

    J 38

    16

    7

    53

    53

    23

    136

    12

    5

    2

  • 113

    4

    1

    13

    95

    29

    51

    27

    12

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    13

    67

    41

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    39

    138

    7

    102

  • 114

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    ilocisolip

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    200901135_B.aalborgi

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    2011

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    2009

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    102

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    B256_B

    .innocens200101917_B.aalborgi

    200901

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    sini

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    200608343_B.hominis

    201116007_B.hominis

    201007259_

    B.aalborgi

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    0 . 06

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    201102

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    201122867_B.aal

    borgi

    201050465_B.aalborgi

    200905511_B.hominis

    200602985_

    B.aalborgi

  • 115

    200624827_B.aalborgi

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    i

    iloc

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    B_52

    7421

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    2 00 8

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    2 0 0 8 1 3 34 0 _ B . a a

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    200922746_B. aal bor g

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  • 116

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  • 117

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