Epidemiology and pathogenicity of African bat lyssaviruses Wanda Markotter University of Pretoria South Africa.

Epidemiology and pathogenicity of

African bat lyssaviruses

Wanda MarkotterUniversity of Pretoria

South Africa

Irkut

WCBVKhujand

RABV (gt1)

ABLV (gt7)

MOKV (gt3)

LBV (gt2)

DUVV (gt4)

EBLV-1 (gt5)

EBLV-2 (gt6)

Aravan

Phylogroup II

Phylogroup IPhylogroupIII

19702006

1981

DUVV (GT 4)

1986

3 isolates from South Africa

19702006

1981

DUVV (GT 4)

1986

3 isolates from South Africa

Paweska et al., 2006. Fatal human infection with rabies-related Duvenhage virus, South Africa. Emerg Infect Dis.2006;12:1965-1967.

LBV isolates

1956Nigeria

Central African Republic1974

South Africa1980(3 isolates)1982

Senegal1985

Zimbabwe1986

1990

Ethiopia1989-1990

Egypt1999

Guinea1985

?

RABV(M13215)

RABV(M31046)

RABV(U22482)

RABV(U42707)

RABV(D42112)

RABV(U22656)

RABV(AF467949)

RABV(U22486)

RABV(U27221)

RABV(AF351834)

ABLV(AF006497)

ABLV(AF081020)

EBLV2(U22847)

EBLV2(U22846)

Khujand(AY262024)

DUVV(AY996324)

DUVV(U22848)

Aravan(AY2620323)

Irkut(AY333112)

EBLV1(U22845)

EBLV1(U22844)

MOKVZIM(S59448)

MOKVETH(AY333111)

LBVNIG(U22842)

LagSA2004(DQ499945)

LagSA2003(DQ499944)

LagSA2005(DQ201180)

Mongoose2004(DQ499948)

LagSA2006(EF525668)

LBVETH(AY333110)

LagSA2003canine(EF525669)

WCBV(AY333113)

100

100

100

42

74

59

100

100

100

99

47

96

55

41

93

80

100

95

86

57

65

59

42

57

56

0.05

Several new LBV isolates in 2003-2006

from South Africa

4 isolates from fruit bats

RABV(M13215)

RABV(M31046)

RABV(U22482)

RABV(U42707)

RABV(D42112)

RABV(U22656)

RABV(AF467949)

RABV(U22486)

RABV(U27221)

RABV(AF351834)

ABLV(AF006497)

ABLV(AF081020)

EBLV2(U22847)

EBLV2(U22846)

Khujand(AY262024)

DUVV(AY996324)

DUVV(U22848)

Aravan(AY2620323)

Irkut(AY333112)

EBLV1(U22845)

EBLV1(U22844)

MOKVZIM(S59448)

MOKVETH(AY333111)

LBVNIG(U22842)

LagSA2004(DQ499945)

LagSA2003(DQ499944)

LagSA2005(DQ201180)

Mongoose2004(DQ499948)

LagSA2006(EF525668)

LBVETH(AY333110)

LagSA2003canine(EF525669)

WCBV(AY333113)

100

100

100

42

74

59

100

100

100

99

47

96

55

41

93

80

100

95

86

57

65

59

42

57

56

0.05

Several new LBV isolates in 2003-2006

from South Africa

4 isolates from fruit bats

Markotter W, Randles J, Rupprecht CE, Sabeta CT, Wandeler AI, Taylor PJ, Nel LH.Recent Lagos bat virus isolations from bats (suborder Megachiroptera) in South Africa. Emerg Infect Dis. 2006;12:504-506.

RABV(M13215)

RABV(M31046)

RABV(U22482)

RABV(U42707)

RABV(D42112)

RABV(U22656)

RABV(AF467949)

RABV(U22486)

RABV(U27221)

RABV(AF351834)

ABLV(AF006497)

ABLV(AF081020)

EBLV2(U22847)

EBLV2(U22846)

Khujand(AY262024)

DUVV(AY996324)

DUVV(U22848)

Aravan(AY2620323)

Irkut(AY333112)

EBLV1(U22845)

EBLV1(U22844)

MOKVZIM(S59448)

MOKVETH(AY333111)

LBVNIG(U22842)

LagSA2004(DQ499945)

LagSA2003(DQ499944)

LagSA2005(DQ201180)

Mongoose2004(DQ499948)

LagSA2006(EF525668)

LBVETH(AY333110)

LagSA2003canine(EF525669)

WCBV(AY333113)

100

100

100

42

74

59

100

100

100

99

47

96

55

41

93

80

100

95

86

57

65

59

42

57

56

0.05

1 isolate from a mongoose

Several new LBV isolates in 2003-2006 from South Africa

First isolation of LBV from terrestrial wildlife

RABV(M13215)

RABV(M31046)

RABV(U22482)

RABV(U42707)

RABV(D42112)

RABV(U22656)

RABV(AF467949)

RABV(U22486)

RABV(U27221)

RABV(AF351834)

ABLV(AF006497)

ABLV(AF081020)

EBLV2(U22847)

EBLV2(U22846)

Khujand(AY262024)

DUVV(AY996324)

DUVV(U22848)

Aravan(AY2620323)

Irkut(AY333112)

EBLV1(U22845)

EBLV1(U22844)

MOKVZIM(S59448)

MOKVETH(AY333111)

LBVNIG(U22842)

LagSA2004(DQ499945)

LagSA2003(DQ499944)

LagSA2005(DQ201180)

Mongoose2004(DQ499948)

LagSA2006(EF525668)

LBVETH(AY333110)

LagSA2003canine(EF525669)

WCBV(AY333113)

100

100

100

42

74

59

100

100

100

99

47

96

55

41

93

80

100

95

86

57

65

59

42

57

56

0.05

1 isolate from a mongoose

Several new LBV isolates in 2003-2006 from South Africa

First isolation of LBV from terrestrial wildlife

Markotter W, Kuzmin I, Rupprecht CE, Randles J, Sabeta CT, Wandeler AI, Nel LH. Isolation of Lagos bat virus from water mongoose. Emerg Infect Dis. 2006;12:1913-1918.

RABV(M13215)

RABV(M31046)

RABV(U22482)

RABV(U42707)

RABV(D42112)

RABV(U22656)

RABV(AF467949)

RABV(U22486)

RABV(U27221)

RABV(AF351834)

ABLV(AF006497)

ABLV(AF081020)

EBLV2(U22847)

EBLV2(U22846)

Khujand(AY262024)

DUVV(AY996324)

DUVV(U22848)

Aravan(AY2620323)

Irkut(AY333112)

EBLV1(U22845)

EBLV1(U22844)

MOKVZIM(S59448)

MOKVETH(AY333111)

LBVNIG(U22842)

LagSA2004(DQ499945)

LagSA2003(DQ499944)

LagSA2005(DQ201180)

Mongoose2004(DQ499948)

LagSA2006(EF525668)

LBVETH(AY333110)

LagSA2003canine(EF525669)

WCBV(AY333113)

100

100

100

42

74

59

100

100

100

99

47

96

55

41

93

80

100

95

86

57

65

59

42

57

56

0.05

1 isolate from a dog

Several new LBV isolates in 2003-2006 from South Africa

11 isolates from South Africa

LBV

Total: 18 isolates

N gene analysis

LBV

Gt 3

Gt 2

WCBV

Lineage C

Lineage B

Lineage A

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA198290

71

78

99

100100

100100

100

98

88

93

0.05

Gt 1

Gt 7

Gt 6

Gt 4

Gt 5

Gt 3

Gt 2

WCBV

Irkut

Aravan

Khujand

Lineage C

Lineage B

Lineage A

AY170412 AY170413 AY170414 AY170415 AY170404 AF351850 AF045166 AF351852 AF351841 AY705373 AF394883 AF394884

AF351833 AY039228 RVU22640 RVU22653 RVU22628

AB085828 RAVRABCVS RAVCGA

AB044824 AY573937 AY573965 AY573949 NC 003243

AF006497 AY573948 AY573958 AY573955 AY573956 AY262023

AY863405 RVU22847 AY863404 AY863407

AY863408 AY863406

AY262024 AY333112 AY996323 AY996324

AY863369 AY863381 AY863398 AY863401 AY863390 AY863385 AY863387 AY863388 AY863383

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA1982

938282

100

100

99

100

100

100

5887

100

94

70

81

100

69100

90

71

78

99

100100

100100

100

100

9996

58

70

100

91

7399

100

8199

100

100

100

57

68

100

100

100

56

79

73

32

80

79

99

9888

93

0.05

Gt 3

Gt 2

WCBV

Lineage C

Lineage B

Lineage A

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA198290

71

78

99

100100

100100

100

98

88

93

0.05

Gt 1

Gt 7

Gt 6

Gt 4

Gt 5

Gt 3

Gt 2

WCBV

Irkut

Aravan

Khujand

Lineage C

Lineage B

Lineage A

AY170412 AY170413 AY170414 AY170415 AY170404 AF351850 AF045166 AF351852 AF351841 AY705373 AF394883 AF394884

AF351833 AY039228 RVU22640 RVU22653 RVU22628

AB085828 RAVRABCVS RAVCGA

AB044824 AY573937 AY573965 AY573949 NC 003243

AF006497 AY573948 AY573958 AY573955 AY573956 AY262023

AY863405 RVU22847 AY863404 AY863407

AY863408 AY863406

AY262024 AY333112 AY996323 AY996324

AY863369 AY863381 AY863398 AY863401 AY863390 AY863385 AY863387 AY863388 AY863383

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA1982

938282

100

100

99

100

100

100

5887

100

94

70

81

100

69100

90

71

78

99

100100

100100

100

100

9996

58

70

100

91

7399

100

8199

100

100

100

57

68

100

100

100

56

79

73

32

80

79

99

98

88

93

0.05

94.8-99.9%

99.3%

Intrinsic variation

Gt 3

Gt 2

WCBV

Lineage C

Lineage B

Lineage A

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA198290

71

78

99

100100

100100

100

98

88

93

0.05

Gt 1

Gt 7

Gt 6

Gt 4

Gt 5

Gt 3

Gt 2

WCBV

Irkut

Aravan

Khujand

Lineage C

Lineage B

Lineage A

AY170412 AY170413 AY170414 AY170415 AY170404 AF351850 AF045166 AF351852 AF351841 AY705373 AF394883 AF394884

AF351833 AY039228 RVU22640 RVU22653 RVU22628

AB085828 RAVRABCVS RAVCGA

AB044824 AY573937 AY573965 AY573949 NC 003243

AF006497 AY573948 AY573958 AY573955 AY573956 AY262023

AY863405 RVU22847 AY863404 AY863407

AY863408 AY863406

AY262024 AY333112 AY996323 AY996324

AY863369 AY863381 AY863398 AY863401 AY863390 AY863385 AY863387 AY863388 AY863383

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA1982

938282

100

100

99

100

100

100

5887

100

94

70

81

100

69100

90

71

78

99

100100

100100

100

100

9996

58

70

100

91

7399

100

8199

100

100

100

57

68

100

100

100

56

79

73

32

80

79

99

98

88

93

0.05

Between B and C 82.7-83.3%

Between A and B,C 79.1-80.7%

Gt 3

Gt 2

WCBV

Lineage C

Lineage B

Lineage A

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA198290

71

78

99

100100

100100

100

98

88

93

0.05

Gt 1

Gt 7

Gt 6

Gt 4

Gt 5

Gt 3

Gt 2

WCBV

Irkut

Aravan

Khujand

Lineage C

Lineage B

Lineage A

AY170412 AY170413 AY170414 AY170415 AY170404 AF351850 AF045166 AF351852 AF351841 AY705373 AF394883 AF394884

AF351833 AY039228 RVU22640 RVU22653 RVU22628

AB085828 RAVRABCVS RAVCGA

AB044824 AY573937 AY573965 AY573949 NC 003243

AF006497 AY573948 AY573958 AY573955 AY573956 AY262023

AY863405 RVU22847 AY863404 AY863407

AY863408 AY863406

AY262024 AY333112 AY996323 AY996324

AY863369 AY863381 AY863398 AY863401 AY863390 AY863385 AY863387 AY863388 AY863383

AY333113 MVU22843

Y09762 AY333111

LBVAFR1999 LBVSEN1985) LBVNIG1956

LBVCAR1974 LBVZIM1986 LagSA2004 LagSA2003 LBVSA2006) Mongoose2004 LBVSA1980(1248) LBVSA1980(640) LBVSA1980(679) LBVSA1982

938282

100

100

99

100

100

100

5887

100

94

70

81

100

69100

90

71

78

99

100100

100100

100

100

9996

58

70

100

91

7399

100

8199

100

100

100

57

68

100

100

100

56

79

73

32

80

79

99

98

88

93

0.05

Between B and C 82.7-83.3%

Between A and B,C 79.1-80.7%

<80% nt identity

? New genotype

0.80.9

11.11.21.31.41.51.61.7

Gene

Min

imum

intr

agen

otyp

ic

iden

tity/

Max

imum

inte

rgen

otyp

ic

iden

tity

Lineage A

Lineage A-C

Lineage B and C

A value of < 1 indicates an overlap.

Irk u t

W C B VK h u ja n d

R A B V (g t1 )

A B LV (g t7 )

M O K V (g t3 )

L B V (g t2 )

D U V V (g t4 )

E B LV-1 (g t5 )

E B LV-2 (g t6 )

A ra v a n

Phylogroup II

Phylogroup I

Pathogenicity

i.m. and i.c.

i.c.

Intracerebral (i.c.) and intramuscular (i.m.) inoculation

020406080

100

Virus isolate

Dea

ths

(%) A

B

C

1 x 102 LD50

1 x 103 LD50

1 x 106 LD50

Conclusions

• Both DUVV and LBV is endemic in South Africa but not frequently reported due to lack of surveillance

• Genetic diversity of LBV has been underestimated• Molecular phylogeny divided previously reported LBV isolates into

three lineages independent of the gene used.• Two isolates previously classified as LBV fulfill the criteria to be a

new lyssavirus genotype• Pathogenic profiles of phylogroup II isolates differ and some isolates

indicate the same pathogenicity as a gt 1 representatives• Pathogenic characteristics of lyssavirus genotypes or phylogroups

should not be decided based on one representative isolate

Acknowledgements

• Prof. Louis H. Nel (UP, South Africa)• Dr. Charles E. Rupprecht (CDC, USA)• Dr. Ivan Kuzmin (CDC, USA)• Dr. Claude T. Sabeta (OVI, South Africa)• Dr. Anthony R. Fooks (VLA, UK)• Dr. Florence Cliquet (AFFSA, France)• Dr. Janusz T Paweska (Special Pathogens, NICD, South Africa)• Dr. Robert Swanepoel (Special Pathogens, NICD, South Africa)• Miss. Charmaine Wilsenach (UP, South Africa)