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A Century of Shope Papillomavirus in Museum Rabbit Specimens.

Escudero Duch C, Williams RA, Timm RM, Perez-Tris J, Benitez L - PLoS ONE (2015)

Bottom Line: Prevalence of symptomatic individuals was significantly higher in Sylvilagus (1.8%) than Lepus.Using multiple methodologies, rolling circle amplification and, multiple isothermal displacement amplification in addition to PCR, greatly improved detection rates.Our results do not clarify whether strains are host species specific, though they are consistent with SfPV specificity to genus Sylvilagus.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology III, Faculty of Biological Sciences, Universidad Complutense de Madrid, Madrid, Spain.

ABSTRACT
Sylvilagus floridanus Papillomavirus (SfPV) causes growth of large horn-like tumors on rabbits. SfPV was described in cottontail rabbits (probably Sylvilagus floridanus) from Kansas and Iowa by Richard Shope in 1933, and detected in S. audubonii in 2011. It is known almost exclusively from the US Midwest. We explored the University of Kansas Natural History Museum for historical museum specimens infected with SfPV, using molecular techniques, to assess if additional wild species host SfPV, and whether SfPV occurs throughout the host range, or just in the Midwest. Secondary aims were to detect distinct strains, and evidence for strain spatio-temporal specificity. We found 20 of 1395 rabbits in the KU collection SfPV symptomatic. Three of 17 lagomorph species (S. nuttallii, and the two known hosts) were symptomatic, while Brachylagus, Lepus and eight additional Sylvilagus species were not. 13 symptomatic individuals were positive by molecular testing, including the first S. nuttallii detection. Prevalence of symptomatic individuals was significantly higher in Sylvilagus (1.8%) than Lepus. Half of these specimens came from Kansas, though new molecular detections were obtained from Jalisco-Mexico's first-and Nebraska, Nevada, New Mexico, and Texas, USA. We document the oldest lab-confirmed case (Kansas, 1915), pre-dating Shope's first case. SfPV amplification was possible from 63.2% of symptomatic museum specimens. Using multiple methodologies, rolling circle amplification and, multiple isothermal displacement amplification in addition to PCR, greatly improved detection rates. Short sequences were obtained from six individuals for two genes. L1 gene sequences were identical to all previously detected sequences; E7 gene sequences, were more variable, yielding five distinct SfPV1 strains that differing by less than 2% from strains circulating in the Midwest and Mexico, between 1915 and 2005. Our results do not clarify whether strains are host species specific, though they are consistent with SfPV specificity to genus Sylvilagus.

No MeSH data available.


Related in: MedlinePlus

Maximum likelihood tree of partial E7 SfPV 1 sequences.The tree was constructed using six 153 bp E7 sequences obtained in our study (1R, 2R, etc.), supplemented with six sequences from previous studies (GenBank accession numbers shown), along with locality and date. Host species is indicated by color (blue = S. audubonii, red = S. floridanus, white = Sylvilagus sp.). Numbers indicate bootstrap support for internal nodes (with 1000 repetitions). Partial sequence 16R was manually added to its closest relatives (resolved sequence was 100% identical to K02708 and AJ404003), but it was excluded from the bootstrap analysis. The assumed position of sequence 16R is represented with a dashed line.
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pone.0132172.g003: Maximum likelihood tree of partial E7 SfPV 1 sequences.The tree was constructed using six 153 bp E7 sequences obtained in our study (1R, 2R, etc.), supplemented with six sequences from previous studies (GenBank accession numbers shown), along with locality and date. Host species is indicated by color (blue = S. audubonii, red = S. floridanus, white = Sylvilagus sp.). Numbers indicate bootstrap support for internal nodes (with 1000 repetitions). Partial sequence 16R was manually added to its closest relatives (resolved sequence was 100% identical to K02708 and AJ404003), but it was excluded from the bootstrap analysis. The assumed position of sequence 16R is represented with a dashed line.

Mentions: The viral L1 sequence (179 bp) from isolates with the stronger signal (1R, 2R, 3R, and 16R) corresponding to S. floridanus showed 100% identity to previous published SfPV1a sequences [11,19,20], all from Kansas. In contrast, E7 sequence (153 bp, excluding primer sequences) from isolates 1R, 2R, 3R, 6R, 7R, and 16R showed some differences (Fig 3 and S5 Fig). Partial E7 sequences generated in this study, below the minimum sequence length for accession to GenBank, are shown in S1 and S2 Tables. Both AIC and BIC selected Jukes–Cantor (JC69) as the best model of nucleotide substitution for the alignment of E7 sequences, with a significant proportion of invariable sites. The maximum likelihood tree (built with the JC+i model of nucleotide substitution) showed low support for most internal nodes, as was expected due to the short length of the alignment, but still allowed the provisional placement of sequences obtained in this study into a broader phylogenetic framework. Sequences 1R and 7R from northcentral (1955) and southeastern (1915) Kansas share 100% identity. The 3R and 6R sequences (both southcentral Nebraska, 2005) were most closely related to one another and grouped with JF303889 (Hershey strain) [20] from Kansas, 1980s (county not provided; identity > 99%), and the identical U09494 (Whidbey Island, Washington state, 1962; USNM 567614). The resolved region of 16R (southwestern Kansas 1994; just 131 bp), was identical to AJ404003 (a4 strain), from southcentral Kansas (1983 [21]), and CTPV “Shope” K02708 [19], also southcentral Kansas (collected at some point between 1966–1977, pers. comm. G. Orth, 30 October 2014). 2R (Jalisco, Mexico, 1966) showed similarity (98.7%) to this group. The subtype b sequence SfPV1 type B AJ243287 [11], also southcentral Kansas (1983) and KC797688, from southeastern Colorado (from S. audubonii, 2010 [12]) are identical. It is noticeable that each sister group occurs within a 45 year period, although the topology of the tree implicates the presence of representatives of each major virus lineage throughout the study period. One clade has not been observed since 1955, a second has not been observed since 1994, while the other two have been observed in the last decade. The three recent clades overlap temporally. All clades occur sympatrically, in Kansas at least. Three groups are relatively close geographically, locality of detection is ca < 600 km (Cherokee and Smith counties, Kansas; Dawson County, Nebraska, and Kansas (county unknown); Kingman County, Kansas and Larimer County, Colorado). Only the identical (for this partial E7 sequence) K02708 and AJ404003, both from southcentral Kansas, and 3R and 6R from southeastern Colorado, were geographically closest to the sequence with which they shared highest identity. The remaining group contains cases from Grant and Kingman counties, Kansas and Jalisco, Mexico, which are approximately 1900 km apart.


A Century of Shope Papillomavirus in Museum Rabbit Specimens.

Escudero Duch C, Williams RA, Timm RM, Perez-Tris J, Benitez L - PLoS ONE (2015)

Maximum likelihood tree of partial E7 SfPV 1 sequences.The tree was constructed using six 153 bp E7 sequences obtained in our study (1R, 2R, etc.), supplemented with six sequences from previous studies (GenBank accession numbers shown), along with locality and date. Host species is indicated by color (blue = S. audubonii, red = S. floridanus, white = Sylvilagus sp.). Numbers indicate bootstrap support for internal nodes (with 1000 repetitions). Partial sequence 16R was manually added to its closest relatives (resolved sequence was 100% identical to K02708 and AJ404003), but it was excluded from the bootstrap analysis. The assumed position of sequence 16R is represented with a dashed line.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493010&req=5

pone.0132172.g003: Maximum likelihood tree of partial E7 SfPV 1 sequences.The tree was constructed using six 153 bp E7 sequences obtained in our study (1R, 2R, etc.), supplemented with six sequences from previous studies (GenBank accession numbers shown), along with locality and date. Host species is indicated by color (blue = S. audubonii, red = S. floridanus, white = Sylvilagus sp.). Numbers indicate bootstrap support for internal nodes (with 1000 repetitions). Partial sequence 16R was manually added to its closest relatives (resolved sequence was 100% identical to K02708 and AJ404003), but it was excluded from the bootstrap analysis. The assumed position of sequence 16R is represented with a dashed line.
Mentions: The viral L1 sequence (179 bp) from isolates with the stronger signal (1R, 2R, 3R, and 16R) corresponding to S. floridanus showed 100% identity to previous published SfPV1a sequences [11,19,20], all from Kansas. In contrast, E7 sequence (153 bp, excluding primer sequences) from isolates 1R, 2R, 3R, 6R, 7R, and 16R showed some differences (Fig 3 and S5 Fig). Partial E7 sequences generated in this study, below the minimum sequence length for accession to GenBank, are shown in S1 and S2 Tables. Both AIC and BIC selected Jukes–Cantor (JC69) as the best model of nucleotide substitution for the alignment of E7 sequences, with a significant proportion of invariable sites. The maximum likelihood tree (built with the JC+i model of nucleotide substitution) showed low support for most internal nodes, as was expected due to the short length of the alignment, but still allowed the provisional placement of sequences obtained in this study into a broader phylogenetic framework. Sequences 1R and 7R from northcentral (1955) and southeastern (1915) Kansas share 100% identity. The 3R and 6R sequences (both southcentral Nebraska, 2005) were most closely related to one another and grouped with JF303889 (Hershey strain) [20] from Kansas, 1980s (county not provided; identity > 99%), and the identical U09494 (Whidbey Island, Washington state, 1962; USNM 567614). The resolved region of 16R (southwestern Kansas 1994; just 131 bp), was identical to AJ404003 (a4 strain), from southcentral Kansas (1983 [21]), and CTPV “Shope” K02708 [19], also southcentral Kansas (collected at some point between 1966–1977, pers. comm. G. Orth, 30 October 2014). 2R (Jalisco, Mexico, 1966) showed similarity (98.7%) to this group. The subtype b sequence SfPV1 type B AJ243287 [11], also southcentral Kansas (1983) and KC797688, from southeastern Colorado (from S. audubonii, 2010 [12]) are identical. It is noticeable that each sister group occurs within a 45 year period, although the topology of the tree implicates the presence of representatives of each major virus lineage throughout the study period. One clade has not been observed since 1955, a second has not been observed since 1994, while the other two have been observed in the last decade. The three recent clades overlap temporally. All clades occur sympatrically, in Kansas at least. Three groups are relatively close geographically, locality of detection is ca < 600 km (Cherokee and Smith counties, Kansas; Dawson County, Nebraska, and Kansas (county unknown); Kingman County, Kansas and Larimer County, Colorado). Only the identical (for this partial E7 sequence) K02708 and AJ404003, both from southcentral Kansas, and 3R and 6R from southeastern Colorado, were geographically closest to the sequence with which they shared highest identity. The remaining group contains cases from Grant and Kingman counties, Kansas and Jalisco, Mexico, which are approximately 1900 km apart.

Bottom Line: Prevalence of symptomatic individuals was significantly higher in Sylvilagus (1.8%) than Lepus.Using multiple methodologies, rolling circle amplification and, multiple isothermal displacement amplification in addition to PCR, greatly improved detection rates.Our results do not clarify whether strains are host species specific, though they are consistent with SfPV specificity to genus Sylvilagus.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology III, Faculty of Biological Sciences, Universidad Complutense de Madrid, Madrid, Spain.

ABSTRACT
Sylvilagus floridanus Papillomavirus (SfPV) causes growth of large horn-like tumors on rabbits. SfPV was described in cottontail rabbits (probably Sylvilagus floridanus) from Kansas and Iowa by Richard Shope in 1933, and detected in S. audubonii in 2011. It is known almost exclusively from the US Midwest. We explored the University of Kansas Natural History Museum for historical museum specimens infected with SfPV, using molecular techniques, to assess if additional wild species host SfPV, and whether SfPV occurs throughout the host range, or just in the Midwest. Secondary aims were to detect distinct strains, and evidence for strain spatio-temporal specificity. We found 20 of 1395 rabbits in the KU collection SfPV symptomatic. Three of 17 lagomorph species (S. nuttallii, and the two known hosts) were symptomatic, while Brachylagus, Lepus and eight additional Sylvilagus species were not. 13 symptomatic individuals were positive by molecular testing, including the first S. nuttallii detection. Prevalence of symptomatic individuals was significantly higher in Sylvilagus (1.8%) than Lepus. Half of these specimens came from Kansas, though new molecular detections were obtained from Jalisco-Mexico's first-and Nebraska, Nevada, New Mexico, and Texas, USA. We document the oldest lab-confirmed case (Kansas, 1915), pre-dating Shope's first case. SfPV amplification was possible from 63.2% of symptomatic museum specimens. Using multiple methodologies, rolling circle amplification and, multiple isothermal displacement amplification in addition to PCR, greatly improved detection rates. Short sequences were obtained from six individuals for two genes. L1 gene sequences were identical to all previously detected sequences; E7 gene sequences, were more variable, yielding five distinct SfPV1 strains that differing by less than 2% from strains circulating in the Midwest and Mexico, between 1915 and 2005. Our results do not clarify whether strains are host species specific, though they are consistent with SfPV specificity to genus Sylvilagus.

No MeSH data available.


Related in: MedlinePlus