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Journal of Clinical Microbiology, August 2007, p. 2751-2753, Vol. 45, No. 8
0095-1137/07/$08.00+0     doi:10.1128/JCM.00230-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

LETTER TO THE EDITOR

Evidence for Bovine Origin of VP4 and VP7 Genes of Human Group A Rotavirus G6P[14] and G10P[14] Strains{triangledown}


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Earlier studies have proposed that human G6P[14] (PA169-like VP7), G8P[14], and G10P[14] group A rotavirus strains resulted from interspecies transmission between humans and cattle (1, 4, 9, 11, 13, 14). However, to our knowledge, PA169-like G6 strains have not been detected in cattle, and the only reported bovine P[14] strain, Sun9, shared borderline VP8* amino acid identities (88.3% to 90.3%) with human P[14] rotaviruses (3) and from phylogenetic analysis appeared to be distantly related to human P[14] strains. In the present study, we report the detection of PA169-like G6 with P[11] and G10P[14] strains (closely related to human P[14] rotaviruses) in calves with diarrhea.

Multiplex seminested PCR using type-specific primers described earlier (2, 5-8, 10) and/or sequencing-based genotyping of 130 (22.7%) group A rotavirus strains (designated RUBV strains for ruminant [RU] and bovine [BV]) detected by RNA electrophoresis in polyacrylamide gels from diarrheic calves (n = 573) from November 2001 to March 2005 in and around Kolkata in eastern India revealed an increased prevalence of G6P[11] strains (89.2%) (Table 1). All 116 RUBV G6P[11] strains exhibited identical electropherotypes in polyacrylamide gels, and 23 of these strains, detected during different time periods of the year and scattered over the entire surveillance period, were sequenced for the VP7 and VP8* genes. Sequence analysis of the VP7 gene of the 23 G6 strains revealed two distinct G6 lineages, with 14 bovine rotavirus strains exhibiting amino acid identities of 95.2% to 98.0% with bovine G6P[11] strain VMRI-29 and 9 bovine rotavirus strains exhibiting amino acid identities of 96.0% to 98.8% with human G6P[14] strain Hun5. This was confirmed by phylogenetic analysis, where representative RUBV strains related to VMRI-29 (RUBV118, RUBV273, and RUBV287) and Hun5 (RUBV28, RUBV315, and RUBV319) clustered within the conserved G6P[11] bovine MC27-like and G6P[14] human PA169-like lineages (Fig. 1A). On the other hand, sequence analysis of the VP8* genes of the 23 RUBV G6P[11] strains revealed maximum amino acid identities (94.9% to 97.3%) with the bovine G10P[11] strain B223, and this genetic relatedness was corroborated by phylogenetic analysis of the VP8* amino acid sequences of RUBV strains related to the MC27-like and PA169-like G6 lineages (Fig. 1B). Phylogenetic analysis of the VP6 and NSP4 genes of a representative strain (RUBV319) pointed towards the bovine origin of PA169-like RUBV strains (data not shown).


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  		TABLE 1. Distribution of G and P genotypes of bovine group A rotavirus strains detected between November 2001 and March 2005 from diarrheic calves in and around Kolkata, West Bengal, India

 

Figure 1
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  		FIG. 1. (A) Phylogenetic analysis of the deduced VP7 amino acid sequences of G6P[11] RUBV strains with representative G6 strains. The five G6 lineages are bracketed at the right. The positions of PA169-like and MC27-like RUBV strains have been indicated with a brace. The tree was rooted with the cognate sequence of G3P[9] strain AU-1. (B) Phylogenetic analysis of the VP8* amino acid sequence of RUBV G6P[11] strains (PA169-like and MC27-like) with strains representing the 27 P genotypes. The other P[11] strains used in the analysis were from earlier published data, and their G genotypes have been mentioned. The cluster comprising RUBV strains has been delineated with a brace. The tree was rooted with the cognate sequence of the human group C rotavirus Bristol strain. Both trees were constructed using the neighbor-joining tree method (random number generator seed of 111 and 1,000 bootstrap trials).

 
Contrary to observations from other parts of the country (12), the prevalence of G10 strains was unusually low (4.6%) (Table 1). Interestingly, the G10P[14] strain RUBV81 shared maximum VP7 deduced amino acid identities of 97.5% and 96.2% with bovine G10P[11] strain B8 and human G10P[14] strain Mc35, respectively. By phylogenetic analysis, strains RUBV81, B8, and Mc35 clustered together (Fig. 2A). The VP8* portion of the VP4 gene of RUBV81 shared maximum amino acid identities of 92.7%, 95.5% to 96.8%, and 96.4% to 97.9% with cognate sequences of the human G10P[14], G8P[14], and G6P[14] strains, respectively, and exhibited amino acid identities of 90.7% and 91.1% to 92.3% with the bovine G8P[14] strain Sun9 and lapine P[14] strains, respectively. By phylogenetic analysis, RUBV81 clustered with human G6P[14], G8P[14], and G10P[14] strains (Fig. 2B). The bovine origin of RUBV81 was confirmed by phylogenetic analyses of its VP6 and NSP4 genes (data not shown).


Figure 2
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  		FIG. 2. (A) Phylogenetic analysis of the deduced VP7 amino acid sequence of G10P[14] strain RUBV81 with representative G10 strains. The tree was rooted with the cognate sequence of G3P[9] strain AU-1. (B) Clustering of the VP8* amino acid sequence of RUBV81 with those of P[14] strains as revealed by phylogenetic analysis. The lineage comprised of human P[14] strains has been marked with a bracket. The tree was rooted with the cognate sequence of the human group C rotavirus Bristol strain. Both trees were constructed using the neighbor-joining tree method (random number generator seed of 111 and 1,000 bootstrap trials).

 
Therefore, the genetic relatedness of RUBV81 to Mc35 provides direct evidence for the bovine origin of the VP4 and VP7 genes of human G10P[14] strains. Similarly, the detection of a bovine P[14] allele closely related to those of human P[14] strains and PA169-like G6 rotaviruses in calves from the same geographical region suggests that G6P[14] strains might have originated from reassortment events involving bovine G6 (PA169-like) and P[14] (RUBV81-like) strains and were subsequently transmitted to humans.

GenBank accession numbers of VP7 and VP4 nucleotide sequences are EF199473 to EF199486 and EF199487 to EF199500, EF199501 to EF199509 and EF199510 to EF199518, and EF200547 and EF200548 for MC27-like G6P[11], PA169-like G6P[11], and G10P[14] RUBV strains, respectively; EF200566 and EF200573 for the VP6 and NSP4 genes of RUBV81, respectively; and EF200571 and EF200578 for the VP6 and NSP4 genes of RUBV319, respectively.


   ACKNOWLEDGMENTS
 
The research was funded by the Department of Biotechnology, Government of India. S. Ghosh, V. Varghese, and S. Samajdar were supported by Senior Research Fellowships from the Indian Council of Medical Research and the Council of Scientific and Industrial Research, Government of India.


   FOOTNOTES
 
{triangledown} Published ahead of print on 30 May 2007. Back


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Souvik Ghosh
Vici Varghese
Sudipta Samajdar
Manju Sinha
Trailokya N. Naik*
Division of Virology
National Institute of Cholera
and Enteric Diseases
Kolkata-700010, India

Nobumichi Kobayashi
Department of Hygiene
Sapporo Medical University
School of Medicine
Sapporo-060, Japan

* Phone: 91-33-23701176, Fax: 91-33-23705066, E-mail: tnaik{at}satyam.net.in; tnnaik{at}gmail.com

Journal of Clinical Microbiology, August 2007, p. 2751-2753, Vol. 45, No. 8
0095-1137/07/$08.00+0     doi:10.1128/JCM.00230-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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