1

A better understanding of the behaviour of different topotypes of Foot-and-Mouth Disease Viruses

circulating in the domestic and wildlife populations.

Dwarka, RM, Maree, FF, Esterhuysen, JJ, Botha, B, Mtshali, N

ARC, Onderstepoort Veterinary Institute, Transboundary Animal Diseases Programme

 

   

   

   

Pool 2   O, A, Asia 1  

Pool 1   O, A, Asia 1

Pool 3   O, A, Asia 1  

The conjectured status of FMD showing approximate distribution of regional virus pools.

Pool 7 O, A

Pool 5 O, A, SAT 1, 2

Pool 4   A, O, SAT 1, 2, 3 Pool 6

SAT 1, 2, 3

Intermediate, sporadic

Endemic

Free

Free. Virus present in game parks

Free with vaccination

Countries with multiples zones: FMD-free, free with vaccination or not free

2

•  In Africa, FMD is widespread throughout the continent

•  Six of the seven serotypes occur on the continent

•  Only serotype Asia 1 not recorded

•  Different ‘patterns’ of disease occur

Vosloo et al., 2002

Map of typed FMD outbreaks in Africa

•  FMD epidemiology is complex on the African continent

•  Serotype distribution differs between geographic regions

•  Wildlife are involved in certain regions

•  Intratypic variants occur within serotypes

Foot-and-mouth disease within pools 4, 5, 6 in Africa

3

Molecular epidemiology of FMD in Africa

•  The genes encoding the structural proteins are used to determine phylogenetic relationships

cccccc

ORF 5’ 3’ UTR UTR

poly A L 1A 1B, 1C, 1D P2 P3

Structural proteins Non-structural proteins

1D: outer capsid coding gene used for initial phylogenetic analysis P1: - Resolve the phylogeny obtained with the 1D gene sequencing

- Greater depth in sequencing information covering the antigenic region of the capsid

- Strains can be selected for vaccine matching studies and structural studies using mathematically modelling Genome length sequencing: Recombination of virus populations

can be studied between/within serotypes

Gene Regions used for sequencing

cccccc

ORF 5’ 3’ UTR UTR

poly A L 1A 1B, 1C, 1D P2 P3

4

Definition of genotypes: group or cluster of closely related viruses When a number of genotypes cluster/group together in a particular geographic location we term this a topotype

Common factors for all serotypes KNP/9/03/Ribye-Waterhole

KNP/1/06/2/Masakosa KNP/06/03/Shangoni SAR/2/04/Selwana-farm

SAR/15/04/Dzumeri SAR/11/01/Craigieburn

KNP/31/95 KNP/07/88 KNP/19/89 KNP/32/92/Boyela

KNP/5/91/Satara SAR/1/03/Masisi

ZIM/08/94 MAL/01/03

MAL/1/08 ZIM/12/02/Chipinge

ZIM/13/02/Chipinge ZIM/6/03/Harare ZIM/4/03/Harare ZIM/7/03/Chinhoyi

MAL/3/75 MAL/01/04/Malawi

ZAM/9/93 ZAM/10/93

ZAM/7/96 BOT/31/98/Vumbura NAM/4/07/Caprivi SWA/4/89

ZIM/10/02/Bikita ZIM/01/01/Bulawayo

ZIM/01/02/Beitbridge BOT/13/02

BOT/4/06/2 BOT/5/06/2

ZIM/34/90 ZIM/267/98/Chizarira

ZIM/7/95/Sengwa NAM/01/92

08/467/Kavango/Namibia BOT/18/98

BOT/1/98/Nxaraga NAM/286/98

ZIM/08/02/Lupane ZIM/07/83/Nyamandlovu

ZIM/4/88 ZIM/42/97

ZIM/48/97 SAU4/00

93 92 99

100

100

100

100

92

100 98

100

99

100

100

92

100

100

100

100

89

81 99

95

87

92

78 100

98 100

78

78

84

61 65

0.05

I

III

II

Database of FMD isolates from buffalo in the KNP KNP/9/03/Ribye-Waterhole

KNP/1/06/2/Masakosa KNP/06/03/Shangoni SAR/2/04/Selwana-farm

SAR/15/04/Dzumeri SAR/11/01/Craigieburn

KNP/31/95 KNP/07/88 KNP/19/89 KNP/32/92/Boyela

KNP/5/91/Satara SAR/1/03/Masisi

ZIM/08/94 MAL/01/03

MAL/1/08 ZIM/12/02/Chipinge

ZIM/13/02/Chipinge ZIM/6/03/Harare ZIM/4/03/Harare ZIM/7/03/Chinhoyi

MAL/3/75 MAL/01/04/Malawi

ZAM/9/93 ZAM/10/93

ZAM/7/96 BOT/31/98/Vumbura NAM/4/07/Caprivi SWA/4/89

ZIM/10/02/Bikita ZIM/01/01/Bulawayo

ZIM/01/02/Beitbridge BOT/13/02

BOT/4/06/2 BOT/5/06/2

ZIM/34/90 ZIM/267/98/Chizarira

ZIM/7/95/Sengwa NAM/01/92

08/467/Kavango/Namibia BOT/18/98

BOT/1/98/Nxaraga NAM/286/98

ZIM/08/02/Lupane ZIM/07/83/Nyamandlovu

ZIM/4/88 ZIM/42/97

ZIM/48/97 SAU4/00

93 92 99

100

100

100

100

92

100 98

100

99

100

100

92

100

100

100

100

89

81 99

95

87

92

78 100

98 100

78

78

84

61 65

0.05

5

SAT - 1 KNP/3/07 Dzombo KNP/5/07 Dzombo KNP/10/07 Bububu KNP/19/07 Langtoon Dam KNP/21/07 Langtoon Dam

SAT - 2 KNP/4/07 Dzombo KNP/11/07 Bububu KNP/20/07 Langtoon Dam KNP/25/07 Boyela

SAT -3 KNP/2/07 Dzombo KNP/26/07 Boyela

Limpopo river

Pafuri Levuvhu river

Olifants river

Punda Maria

Mopani

Shingwedzi

Olifants

Letaba

Phalaborwa

Shingwedzi river

KNP NORTH

Mphongolo

Bububu

Langtoon dam

Boyela

Dzombo Bakoor Shigumane

Ø DZOMBO – SAT-1, SAT-2 & SAT-3 Ø LANGTOON DAM – SAT-1 & SAT-2 Ø BUBUBA – SAT-1 & SAT-2 Ø BOYELA – SAT-2 & SAT-3

KNP isolates updated

SAT -1 Letaba isolates -Eight cluster within

the southern topotype but in four different genotypes

Isolates cluster within the Southern topotype, but in four different genotypes.

Limpopo river Pafuri Levuvhu river

Olifants river

Tim

bava

ti riv

er

Punda Maria

Mopani

Shingwedzi

Olifants Letaba

Satara Klaserie PNR

Sabie Sabie PNR

Timbavati PNR Orpen

Lower Sabie

Crocodile river

Skukuza

Pretoriuskop

Malelane

Shingwedzi river

Crocodile Bridge

Mbyamiti river

KNP/2/06/Maseya KNP/3/06/Maseya

KNP/2/02/Elandskuil KNP/26/08

KNP/893/98/Shipane W/M KNP/35/92/SHINGWEDZI

KNP/6/06/Makwadsi 10/1364/Zidlele DT 10/1693/Feedlot GP

KNP/8/95/MONDSWENI KNP/7/03/Shangoni

KNP/1584/98/Baranukee KNP/8/08 KNP/14/08 KNP/10/07/Bububu KNP/3/07/Dzombo KNP/8/07/Dzombiene

SAR/9/81 GN13/91 KNP/01/01/Shipandi-W/M

SAR/7/03/buffalo SAR/6/03/buffalo

SAR/1/00/Phalaborwa KNP20/89/KWA-MFAMEBTO

SAR/2/10/Phalaborwa SAR/8/10/Gravelotte

KNP/397/98/RIETPAN KNP/400/98/RIETPAN

KNP/22/08 KNP/24/08

KNP/17/96 SAR/08/02/Mauricedale

KNP/21/07/Langtoon Dam KNP/15/08 KNP/11/03/Masorini KNP/18/08 KNP/17/08

ZIM/05/99 SAR/1/09/Makoko DT KNP/5/05 KNP/148/91

53

81

79

99

73 100

100 97

66

100

100 100

100

99

90

100

Phylogenetic analysis SAT 1 KNP viruses

SAT-1 (2008)

I I

II

III

IV

KNP

6

South Africa: Wildlife/ cattle interface

7

Genetic variability of FMDV based on P1 sequencing

SAR1

/200

9

SA

R2/2

009

SA

R4

/20

09

SA

R3/2

009

SAR7

/200

9

SAR5/2009SAR6/2009KNP/196/91/1

KNP/148/91/1SAR/09/81/1KNP/41/95/1ZIM/HV/03/90

ZIM/GN/13/90

ZIM/25/90/1

MOZ/03/02/1

ZAM/02/93/1

TAN/37/99/1

KEN/05/98/1

ZIM/06/94/1

NAM/307/98/1

BOT/3/06/1

UG

A/03/99/1U

GA

/01/97/1N

IG/05/81/1

SU

D/0

3/7

6/1

NIG

/08/76/1N

IG/1

5/7

5/1

NIG

/06/7

6/1

SE

N/0

7/8

3/2

RW

A/0

2/0

1/2

ZA

I/01/7

4/2

UG

A/0

2/0

2/2

KE

N/0

3/5

7/2

SA

U/0

6/0

0/2

ER

I/12/

89/2

GH

A/08

/91/

2

SEN

/05/75

/2

ANG/04

/74/2

ZIM

/14/90

/2

ZIM/17

/91/2

ZIM/34/90/2

BOT/4/06/2

ZIM/07/83/2

ZIM/01/88/2

ZAM/07/96/2 RHO/01/48/2 KEN/08/99/2

ZIM/08/94/2 ZIM/GN/10/91

SAR/16/83/2

KNP/51/93/2

KN

P/02/89/2

KN

P/1

9/8

9/2

UGA/02/97/3

ZAM/04/96/3

BE

C/16/53/3

KN

P/1

0/9

0/3

ZIM

/05/9

1/3

A22

A1

2 O1

Ka

uf

O1T

ai

0.05

SAT1

SAT3

SAT2

Type A Type O

Neighbour-joining tree depicting the relationships between nucleotide sequences of the P1 regions of 5 FMDV serotypes

8

KNP/148/91 [Buffalo]

KNP/ 41/95 [Buffalo]

ZIM/6/94 [Buffalo]

NAM/307/98 [Buffalo]

MOZ/3/02 [Bovine]

KEN/05/98 [Bovine]

UGA/1/97 [Buffalo]

NIG 8/76 [Bovine]

BHK-21 IB-RS2 CHO SAT1

Southern Africa

East Africa West Africa

Plaque morphology assists as a measurement of adaptation to BHK cells

Peninah Nsamba, Belinda. Blignaut and Francois. F. Maree TADP, OVI and University of Pretoria

SAT1 Plaque Phenotypes in different cell lines

SAT2 Plaque Phenotypes in different cell lines

SAT2/KNP 2/89 [Impala]

SAT2/KNP 51/93 [Impala]

SAT2/ZIM 1/88 [Buffalo]

SAT2/ANG/4/74 [Bovine]

SAT2/UGA/2/02 [Bovine]

SAT2/RWA/2/01 [Bovine]

SAT2/SEN/7/83 [Bovine]

SAT2/GHA/8/91 [Bovine]

BHK-21 IB-RS2 CHO SAT2

East Africa

West Africa

Southern Africa

Passage History of virus isolates - PK or BTY – 1-2 times - IB-RS – 3-4 times

Peninah Nsamba, Belinda. Blignaut and Francois. F. Maree TADP, OVI and University of Pretoria

9

Pool 6

Topotype 1

Topotype 3

Topotype 2

SAR/2/09

SAR/4/09

SAR/1/09

SAR/3/09

SAR/7/09

SAR/5/09

SAR/6/09

KNP/196/91

KNP/148/91

KNP/11/03

SAR/8/02

SAR/9/03

SAR/33/00

KNP/41/95

SAR/7/03

KNP/07/03

KNP/10/03

ZIM/HV/03/90

ZIM/GN/13/90

SAR/09/81

KEN/05/98

TAN/37/99

ZAM/01/06

ZAM/02/93

ZIM/03/95

ZIM/14/98

ZIM/25/90

ZIM/11/03

MOZ/01/02

MOZ/03/02

BOT/01/06

BOT/03/06

NAM/01/10

ZIM/03/03

ZIM/06/94

NAM/272/98

BOT/02/98

NAM/307/98

NAM/308/98

UGA/01/97

UGA/03/99

100

99100

99

92

100

69

98

51

86

100

98

100

67

100

77

100

98

100

81

100

100

8384

88

64

100

95

66

69

53

65

100

41

26

72

23

0.05

Vaccine matching studies

Antigenic variation (r values)

•  r-value = heterologous neutralising titer / homologous titer

•  r-value range: 0.4-1.0: good cross-protection against outbreak strain 0.2-0.4: reasonable cross-protection; a potent vaccine will still protect against outbreak virus 0-0.2: poor/no cross-protection

10

Topotype 1 Topotype 2 Topotype 3

ZAM/1/06 SAR/9/81 BOT/1/06 KNP/196/91

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

r-value criteria

SAT2 vaccine strains

SAT2 reference viruses

Per

cent

age

viru

ses

in e

ach

cate

gory

SAT2 antigenic variation

Vaccine match based on r-values

11

Fig. 2; Maree et al.

A B

SAT1/SAR/9/81 SAT2/ZIM/7/83

Structural models of FMDV

Capsid structures of SAT 1 and SAT 2. Variation in surface exposed regions. Study of correlation between the field isolates and vaccines

SAT-1:

Topotype I

Topotype IITopotype III

Topotype IV

Topotype V

Topotype VITopotype VII

Topotype VIII

MORROCCO

MAURITANIA

ALGERIA

GHANA

MALI

GUINEAGUINEA-BISAU

SIERRA-LEONE

LIBERIA

BURKINAFASO

GAMBIA

TOGOBENIN

NIGERIA

NIGER

EGYPT

SOMALIA

LYBIA

CHAD

SUDAN

CAMEROON

CENTRALAFRICANREPUBLIC

ETHIOPIA

EQUATORIALGUINEA UGANDA

RWANDA

TANZANIA

ZAIREGABONCONGO

ANGOLA

NAMIBIA

ZAMBIA

ZIMBABWE

BOTSWANA

SWAZILAND

LESOTHOSOUTHAFRICA

TUNISIA

KENYA

COTED’IVOIRE

SENEGAL

ERITREA

YEMEN

SAUDI ARABIA

ISRAELJORDAN

BURUNDI

12

SAT-2:

Topotype I

Topotype IITopotype III

Topotype IV

Topotype V

Topotype VITopotype VII

Topotype VIIITopotype IX

Topotype XIVTopotype XIIITopotype XII

Topotype XITopotype X

MORROCCO

MAURITANIA

ALGERIA

GHANA

MALI

GUINEA GUINEA- BISAU

SIERRA- LEONE

LIBERIA

BURKINA FASO

GAMBIA TOGO

B E N I N

NIGERIA

NIGER

EGYPT

SOMALIA

LYBIA

CHAD SUDAN

CAMEROON CENTRAL AFRICAN REPUBLIC

ETHIOPIA

EQUATORIAL GUINEA UGANDA

RWANDA

TANZANIA

ZAIRE GABON CONGO

ANGOLA

NAMIBIA

ZAMBIA ZIMBABWE

BOTSWANA SWAZILAND

LESOTHO SOUTH AFRICA

TUNISIA

KENYA

COTE D’IVOIRE

SENEGAL

ERITREA YEMEN

SAUDI ARABIA

ISRAEL JORDAN

BURUNDI

Buffalo/cattle sampling in SADC countries during 2010-2012:

•  Malawi •  Tanzania •  Zambia •  Mozambique •  Angola

Regional Activities SADC TADs Project

13

Concluding remarks •  A comprehensive database of FMD viruses and

sequences from SADC countries enables a better understanding on the evolution of the virus.

•  When a FMD virus database is represented by diverse topotypes, it enables diagnosticians to better understand and trace the possible origins of outbreak strains.

•  Understanding the possible source of an outbreak could play a role in the immediate control strategy for that region. It also enables scientists to assess if a particular topotype is dominant in a region, which could have an impact on vaccine strain selection for that area.

Concluding remarks •  If vaccination is applied as a control measure, a

topotype rich database of viruses will enable better vaccine matching studies to be performed to evaluate the most appropriate vaccine strain.

•  Scientists can study the relationships between FMD viruses from different topotypes and perform vaccine matching studies across topotypes,

•  However, for a holistic regional control strategy, concerted collaborative efforts between governments, industry, farmers and research institutions are central to effectively mitigating the risk posed by a disease such as FMD.

14

Thank You