Jelle Matthijnssens PhD Laboratory of Clinical and Epidemiological Virology Rega Institute for Medical Research University of Leuven, Belgium

DISTRIBUTION AND REASSORTMENT AMONG HUMAN AND ANIMAL ROTAVIRUSES REVEALED BY COMPARATIVE GENOME ANALYSES

11th International Rotavirus Symposium, Delhi, India, September 3-5 2014 SEQUENCE-RBASEDOTAVIRUSCLASSIFICATIONSTRUCTUREFOR 11 GENE SEGMENTS SEQUENCE-BASED CLASSIFICATION FOR 11 GENE SEGMENTS

GENE PRODUCT PERCENTAGE IDENTITY # GENOTYPES GENOTYPE DESIGNATION CUT-OFF VALUES VP7 80% 27 G Glycosylated VP4 80% 37 P Protease sensitive VP6 85% 20 I Inner capsid protein VP1 83% 11 R RNA-dependent RNA-polymerase VP2 84% 11 C Core protein VP3 81% 10 M Methyltransferase NSP1 79% 22 A Interferon Antagonist NSP2 85% 11 N NTPase NSP3 85% 14 T Translation Enhancer NSP4 85% 18 E Enterotoxin NSP5 91% 13 H pHosphoprotein Gx-P[x]-Ix-Rx-Cx-Mx-Ax-Nx-Tx-Ex-Hx COMPLETE ROTAVIRUS GENOMES

• Complete rotavirus genomes in GenBank:

2012 2014

 Human rotaviruses ≈ 400 > 800  Bovine rotaviruses ≈ 20 > 25  Porcine rotaviruses ≈ 15 > 20  Equine rotaviruses ≈ 8 > 10  Canine rotaviruses ≈ 5 > 5 COMPLETE ROTAVIRUS GENOMES

• Complete rotavirus genomes in GenBank:

2012 2014

 Human rotaviruses ≈ 400 > 800  Bovine rotaviruses ≈ 20 > 25  Porcine rotaviruses ≈ 15 > 20  Equine rotaviruses ≈ 8 > 10  Canine rotaviruses ≈ 5 > 5 COMPLETE ROTAVIRUS GENOMES

• Complete rotavirus genomes in GenBank:

2012 2014

 Human rotaviruses ≈ 400 > 800  Bovine rotaviruses ≈ 20 > 25  Porcine rotaviruses ≈ 15 > 20  Equine rotaviruses ≈ 8 > 10  Canine rotaviruses ≈ 5 > 5 GENOTYPE CONSTELLATIONS

Genotype constellation G-genotype P-genotype

VP7 VP4 VP3 VP6 VP1 VP2

NSP1 NSP2 NSP3 NSP4 NSP5 Human Wa-like Gx P[x] I1 R1 C1 M1 A1 N1 T1 E1 H1 G1, G3, G4, G9, G12 P[8], P[6]

Human DS-1-like Gx P[x] I2 R2 C2 M2 A2 N2 T2 E2 H2 G2, G8 P[4], P[6]

Human AU-1-like Gx P[x] I3 R3 C3 M3 A3 N3 T3 E3 H3 G3, G12 P[3],P[9] P[9]

I1 A1 T1 Porcine Gx P[x] R1 C1 M1 N1 E1 H1 G4, G5, G9, G11 P[6],P[7],P[13],P[23] I5 A8 T7

A3 Bovine-like Gx P[x] I2 R2 C2 M2 A11 N2 T6 E2 H3 G6, G8, G10 P[1],P[5],P[11],P[14] A13

I2 E2 Equine Gx P[x] R2 C2 M3 A10 N2 T3 H7 G3, G12G14 P[12] I6 E12

I3 R3 C3 M3 A3 N3 H3 Cat/Dog Gx P[x] T3 E3 G3, G6 P[9] I2 R2 C2 M2 A9 N2 H6

G1 P[5] I2 R2 C2 M1 A3 N2 T6 E2 H3 G2 P[5] I2 R2 C2 M1 A3 N2 T6 E2 H3 G3 P[5] I2 R2 C2 M2 A3 N2 T6 E2 H3 Vaccine strains G4 P[5] I2 R2 C2 M2 A3 N2 T6 E2 H3 G6 P[8] I2 R2 C2 M2 A3 N2 T6 E2 H3 G1 P[8] I1 R1 C1 M1 A1 N1 T1 E1 H1 GENOTYPE CONSTELLATIONS

 Genotype constellations are rather well conserved per host species

 Some genotypes are shared between genotype constellations of different host species GENOTYPE CONSTELLATIONS

 Genotype constellations are rather well conserved per host species

 Some genotypes are shared between genotype constellations of different host species

 Interspecies transmission are frequently described in literature, although usually they represent dead-end infections GENOTYPE CONSTELLATIONS

 Genotype constellations are rather well conserved per host species

 Some genotypes are shared between genotype constellations of different host species

 Interspecies transmission are frequently described in literature, although usually they represent dead-end infections

 Reassortments can occur, resulting in reassortant strains that can circulate in the population GENOTYPE CONSTELLATIONS

 3 examples of rotavirus interspecies transmission and reassortment events GENOTYPE CONSTELLATIONS

 3 examples of rotavirus interspecies transmission and reassortment events

 Animal gene segments in human G2P[4] strains in Belgium GENOTYPE CONSTELLATIONS

 3 examples of rotavirus interspecies transmission and reassortment events

 Animal gene segments in human G2P[4] strains in Belgium

 Unusual bovine-like rotavirus strain in child during rotavirus surveillance in Belgium GENOTYPE CONSTELLATIONS

 3 examples of rotavirus interspecies transmission and reassortment events

 Animal gene segments in human G2P[4] strains in Belgium

 Unusual bovine-like rotavirus strain in child during rotavirus surveillance in Belgium

 Possible interspecies transmission of bat rotaviruses to humans and horses I) G2P[4] IN BELGIUM

 Increased proportion of G2P[4] strains after Rotarix™ introduction in Belgium in 2006 I) G2P[4] IN BELGIUM

 Increased proportion of G2P[4] strains after Rotarix™ introduction in Belgium in 2006

 G2P[4] strains isolated between 1999-2013 were selected based on their phylogenetic clustering of VP7 and VP4 I) G2P[4] IN BELGIUM

 Increased proportion of G2P[4] strains after Rotarix™ introduction in Belgium in 2006

 G2P[4] strains isolated between 1999-2013 were selected based on their phylogenetic clustering of VP7 and VP4

 The NSP4 segment of 89 G2P[4] strains was sequenced

I) G2P[4] IN BELGIUM

 Increased proportion of G2P[4] strains after Rotarix™ introduction in Belgium in 2006

 G2P[4] strains isolated between 1999-2013 were selected based on their phylogenetic clustering of VP7 and VP4

 The NSP4 segment of 89 G2P[4] strains was sequenced

 30 samples were selected for complete genome analyses (ongoing) I) G2P[4] IN BELGIUM

RVA/Human-wt/BEL/17/2002/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/21/2005/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/23/2005/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/24/2005/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/31/2007/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/34/2007/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/38/2007/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H3 RVA/Human-wt/BEL/41/2007/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H3 RVA/Human-wt/BEL/45/2008/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/67/2009/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2 RVA/Human-wt/BEL/75/2010/G2P[4] G2 P[4] I2 R2 C2 M2 A2 N2 T2 E2 H2

 More detailed information can be found on poster P120 II) NEW G/P-GENOTYPES IN HUMAN CHILD

 Rotavirus surveillance in Belgium 2013-2014 II) NEW G/P-GENOTYPES IN HUMAN CHILD

 Rotavirus surveillance in Belgium 2013-2014  Unknown VP7 and VP4 rotavirus sequences detected in an infant II) NEW G/P-GENOTYPES IN HUMAN CHILD

 Rotavirus surveillance in Belgium 2013-2014  Unknown VP7 and VP4 rotavirus sequences detected in an infant  Tentative assigned G28 and P[38] II) NEW G/P-GENOTYPES IN HUMAN CHILD

 Rotavirus surveillance in Belgium 2013-2014  Unknown VP7 and VP4 rotavirus sequences detected in an infant  Tentative assigned G28 and P[38]  Complete genome was determined using Illumina Sequencing

RVA/Human-wt/BEL/F06018/2014/G28P[38] G28 P[38] I2 R2 C2 M2 A11 N2 T6 E2 H3

A3 Bovine-like Gx P[x] I2 R2 C2 M2 A11 N2 T6 E2 H3 A13 III) ROTAVIRUSES IN BATS

RVA/Bat-wt/KEN/KE4582/2007/G25P[6] G25 P[6] I5 C8 N8 T11 E2 H6

RVA/Bat-tc/CHN/MSLH14/2012/G3P[3] G3 P[3] I8 R3 C3 M3 A9 N3 T3 E3 H6 RVA/Bat-tc/CHN/MYAS33/2013/G3P[10] G3 P[10] I8 R3 C3 M3 A9 N3 T3 E3 H6

RVA/Horse-wt/ARG/E3198/2008/G3P[3]EmergJ InfectVirol. 2013Dis.G3 2010P[3] I3 R3 C3 M3 A9 N3 T3 E3 H6 RVA/Human-wt/THA/CMH222/2001/G3P[3] G3 P[3] I8 E3 RVA/Human-wt/THA/CMH079/2005/G3P[10] G3 P[10] I8 E3 H6 Stoliczka trident bat, Unpublished CONCLUSIONS

 Conserved rotavirus genome constellations are present in different host species including humans, despite the constant exposure to animal rotaviruses  Interspecies transmitted viruses usually cause dead- end infections  Reassortant viruses may circulate in the human population for a prolonged period of time  With the decreased circulation of the typical human rotaviruses due to vaccination, unusual animal rotaviruses may have increasing opportunities to infect humans, necessitating continued surveillance CONCLUSIONS

 Conserved rotavirus genome constellations are present in different host species including humans, despite the constant exposure to animal rotaviruses  Interspecies transmitted viruses usually cause dead- end infections  Reassortant viruses may circulate in the human population for a prolonged period of time  With the decreased circulation of the typical human rotaviruses due to vaccination, unusual animal rotaviruses may have increasing opportunities to infect humans, necessitating continued surveillance CONCLUSIONS

 Conserved rotavirus genome constellations are present in different host species including humans, despite the constant exposure to animal rotaviruses  Interspecies transmitted viruses usually cause dead- end infections  Reassortant viruses may circulate in the human population for a prolonged period of time  With the decreased circulation of the typical human rotaviruses due to vaccination, unusual animal rotaviruses may have increasing opportunities to infect humans, necessitating continued surveillance CONCLUSIONS

 Conserved rotavirus genome constellations are present in different host species including humans, despite the constant exposure to animal rotaviruses  Interspecies transmitted viruses usually cause dead- end infections  Reassortant viruses may circulate in the human population for a prolonged period of time  With the decreased circulation of the typical human rotaviruses due to vaccination, unusual animal rotaviruses may have increasing opportunities to infect humans, necessitating continued surveillance ACKNOWLEDGEMENTS

University of Leuven Rega Institute for Medical Research Elisabeth Heylen Mark Zeller Marc Van Ranst

Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun, China Biao He Xia Lele Changchun Tu Jelle Matthijnssens PhD Laboratory of Clinical and Epidemiological Virology Rega Institute for Medical Research University of Leuven, Belgium

DISTRIBUTION AND REASSORTMENT AMONG HUMAN AND ANIMAL ROTAVIRUSES REVEALED BY COMPARATIVE GENOME ANALYSES

11th International Rotavirus Symposium, Delhi, India, September 3-5 2014