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Phylogeny and differentiation of reptilian and amphibian ranaviruses detected in Europe.

Stöhr AC, López-Bueno A, Blahak S, Caeiro MF, Rosa GM, Alves de Matos AP, Martel A, Alejo A, Marschang RE - PLoS ONE (2015)

Bottom Line: In this study, we characterized a number of ranaviruses detected in wild and captive animals in Europe based on sequence data from six genomic regions (major capsid protein (MCP), DNA polymerase (DNApol), ribonucleoside diphosphate reductase alpha and beta subunit-like proteins (RNR-α and -β), viral homolog of the alpha subunit of eukaryotic initiation factor 2, eIF-2α (vIF-2α) genes and microsatellite region).Comparative analyses show that among the closely related amphibian-like ranaviruses (ALRVs) described to date, three recently split and independently evolving distinct genetic groups can be distinguished.These findings underline the wide host range of ranaviruses and the emergence of pathogen pollution via animal trade of ectothermic vertebrates.

View Article: PubMed Central - PubMed

Affiliation: Fachgebiet für Umwelt- und Tierhygiene, Universität Hohenheim, Stuttgart, Germany.

ABSTRACT
Ranaviruses in amphibians and fish are considered emerging pathogens and several isolates have been extensively characterized in different studies. Ranaviruses have also been detected in reptiles with increasing frequency, but the role of reptilian hosts is still unclear and only limited sequence data has been provided. In this study, we characterized a number of ranaviruses detected in wild and captive animals in Europe based on sequence data from six genomic regions (major capsid protein (MCP), DNA polymerase (DNApol), ribonucleoside diphosphate reductase alpha and beta subunit-like proteins (RNR-α and -β), viral homolog of the alpha subunit of eukaryotic initiation factor 2, eIF-2α (vIF-2α) genes and microsatellite region). A total of ten different isolates from reptiles (tortoises, lizards, and a snake) and four ranaviruses from amphibians (anurans, urodeles) were included in the study. Furthermore, the complete genome sequences of three reptilian isolates were determined and a new PCR for rapid classification of the different variants of the genomic arrangement was developed. All ranaviruses showed slight variations on the partial nucleotide sequences from the different genomic regions (92.6-100%). Some very similar isolates could be distinguished by the size of the band from the microsatellite region. Three of the lizard isolates had a truncated vIF-2α gene; the other ranaviruses had full-length genes. In the phylogenetic analyses of concatenated sequences from different genes (3223 nt/10287 aa), the reptilian ranaviruses were often more closely related to amphibian ranaviruses than to each other, and most clustered together with previously detected ranaviruses from the same geographic region of origin. Comparative analyses show that among the closely related amphibian-like ranaviruses (ALRVs) described to date, three recently split and independently evolving distinct genetic groups can be distinguished. These findings underline the wide host range of ranaviruses and the emergence of pathogen pollution via animal trade of ectothermic vertebrates.

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PCR amplification of the microsatellite region from the studied ranavirus isolates and FV3.The amplicons were separated by electrophoresis in 4% agarose gel. Lane 1: 10 bp marker, lane 2: ToRV1 (60 bp), lane 3: ToRV2 (62 bp), lane 4: GGRV (65 bp), lane 5: ACRV (70 bp), lane 6: ASRV (76 bp), lane7: JSpRV (101 bp), lane 8: LMRV (134 bp), lane 9: FV3 (138 bp), lane 10: DGRV (156 bp), lane 11: CH8/96 (164 bp), lane 12: ZPRV1 (230 bp), lane 13: ZPRV2 (288 bp), lane 14: BPRV (351 bp), lane 15: negative control, lane 16: 50 bp marker
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pone.0118633.g002: PCR amplification of the microsatellite region from the studied ranavirus isolates and FV3.The amplicons were separated by electrophoresis in 4% agarose gel. Lane 1: 10 bp marker, lane 2: ToRV1 (60 bp), lane 3: ToRV2 (62 bp), lane 4: GGRV (65 bp), lane 5: ACRV (70 bp), lane 6: ASRV (76 bp), lane7: JSpRV (101 bp), lane 8: LMRV (134 bp), lane 9: FV3 (138 bp), lane 10: DGRV (156 bp), lane 11: CH8/96 (164 bp), lane 12: ZPRV1 (230 bp), lane 13: ZPRV2 (288 bp), lane 14: BPRV (351 bp), lane 15: negative control, lane 16: 50 bp marker

Mentions: The microsatellite region previously described in FV3, STIV, and CMTV was successfully amplified and visualized from all ranavirus isolates (Fig. 2, PNTRV not shown). For most isolates, the size of the amplicons clearly differed from one another (ToRV1: 60 bp, ToRV2: 62 bp, GGRV: 65 bp, ACRV: 70 bp, ASRV: 76 bp, JSpRV: 101 bp, PNTRV: 130 to 140 bp, LMRV: 134 bp, DGRV (Dopasia gracilis ranavirus): 156 bp, CH8/96 (Testudo hermanni ranavirus): 164 bp, ZPRV1 (Zuerich Pelophylax collection ranavirus 1): 230 bp, ZPRV2: 288 bp, BPRV (blood python ranavirus): 351 bp). The single virus which was not isolated in cell culture (NCRV (Neurergus crocatus ranavirus)) could not be visualized. An attempt to sequence the products failed.


Phylogeny and differentiation of reptilian and amphibian ranaviruses detected in Europe.

Stöhr AC, López-Bueno A, Blahak S, Caeiro MF, Rosa GM, Alves de Matos AP, Martel A, Alejo A, Marschang RE - PLoS ONE (2015)

PCR amplification of the microsatellite region from the studied ranavirus isolates and FV3.The amplicons were separated by electrophoresis in 4% agarose gel. Lane 1: 10 bp marker, lane 2: ToRV1 (60 bp), lane 3: ToRV2 (62 bp), lane 4: GGRV (65 bp), lane 5: ACRV (70 bp), lane 6: ASRV (76 bp), lane7: JSpRV (101 bp), lane 8: LMRV (134 bp), lane 9: FV3 (138 bp), lane 10: DGRV (156 bp), lane 11: CH8/96 (164 bp), lane 12: ZPRV1 (230 bp), lane 13: ZPRV2 (288 bp), lane 14: BPRV (351 bp), lane 15: negative control, lane 16: 50 bp marker
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4338083&req=5

pone.0118633.g002: PCR amplification of the microsatellite region from the studied ranavirus isolates and FV3.The amplicons were separated by electrophoresis in 4% agarose gel. Lane 1: 10 bp marker, lane 2: ToRV1 (60 bp), lane 3: ToRV2 (62 bp), lane 4: GGRV (65 bp), lane 5: ACRV (70 bp), lane 6: ASRV (76 bp), lane7: JSpRV (101 bp), lane 8: LMRV (134 bp), lane 9: FV3 (138 bp), lane 10: DGRV (156 bp), lane 11: CH8/96 (164 bp), lane 12: ZPRV1 (230 bp), lane 13: ZPRV2 (288 bp), lane 14: BPRV (351 bp), lane 15: negative control, lane 16: 50 bp marker
Mentions: The microsatellite region previously described in FV3, STIV, and CMTV was successfully amplified and visualized from all ranavirus isolates (Fig. 2, PNTRV not shown). For most isolates, the size of the amplicons clearly differed from one another (ToRV1: 60 bp, ToRV2: 62 bp, GGRV: 65 bp, ACRV: 70 bp, ASRV: 76 bp, JSpRV: 101 bp, PNTRV: 130 to 140 bp, LMRV: 134 bp, DGRV (Dopasia gracilis ranavirus): 156 bp, CH8/96 (Testudo hermanni ranavirus): 164 bp, ZPRV1 (Zuerich Pelophylax collection ranavirus 1): 230 bp, ZPRV2: 288 bp, BPRV (blood python ranavirus): 351 bp). The single virus which was not isolated in cell culture (NCRV (Neurergus crocatus ranavirus)) could not be visualized. An attempt to sequence the products failed.

Bottom Line: In this study, we characterized a number of ranaviruses detected in wild and captive animals in Europe based on sequence data from six genomic regions (major capsid protein (MCP), DNA polymerase (DNApol), ribonucleoside diphosphate reductase alpha and beta subunit-like proteins (RNR-α and -β), viral homolog of the alpha subunit of eukaryotic initiation factor 2, eIF-2α (vIF-2α) genes and microsatellite region).Comparative analyses show that among the closely related amphibian-like ranaviruses (ALRVs) described to date, three recently split and independently evolving distinct genetic groups can be distinguished.These findings underline the wide host range of ranaviruses and the emergence of pathogen pollution via animal trade of ectothermic vertebrates.

View Article: PubMed Central - PubMed

Affiliation: Fachgebiet für Umwelt- und Tierhygiene, Universität Hohenheim, Stuttgart, Germany.

ABSTRACT
Ranaviruses in amphibians and fish are considered emerging pathogens and several isolates have been extensively characterized in different studies. Ranaviruses have also been detected in reptiles with increasing frequency, but the role of reptilian hosts is still unclear and only limited sequence data has been provided. In this study, we characterized a number of ranaviruses detected in wild and captive animals in Europe based on sequence data from six genomic regions (major capsid protein (MCP), DNA polymerase (DNApol), ribonucleoside diphosphate reductase alpha and beta subunit-like proteins (RNR-α and -β), viral homolog of the alpha subunit of eukaryotic initiation factor 2, eIF-2α (vIF-2α) genes and microsatellite region). A total of ten different isolates from reptiles (tortoises, lizards, and a snake) and four ranaviruses from amphibians (anurans, urodeles) were included in the study. Furthermore, the complete genome sequences of three reptilian isolates were determined and a new PCR for rapid classification of the different variants of the genomic arrangement was developed. All ranaviruses showed slight variations on the partial nucleotide sequences from the different genomic regions (92.6-100%). Some very similar isolates could be distinguished by the size of the band from the microsatellite region. Three of the lizard isolates had a truncated vIF-2α gene; the other ranaviruses had full-length genes. In the phylogenetic analyses of concatenated sequences from different genes (3223 nt/10287 aa), the reptilian ranaviruses were often more closely related to amphibian ranaviruses than to each other, and most clustered together with previously detected ranaviruses from the same geographic region of origin. Comparative analyses show that among the closely related amphibian-like ranaviruses (ALRVs) described to date, three recently split and independently evolving distinct genetic groups can be distinguished. These findings underline the wide host range of ranaviruses and the emergence of pathogen pollution via animal trade of ectothermic vertebrates.

Show MeSH