Members in the genus Bocaparvovirus are closely related to human health and have a wide host range. The diverse hosts raise the possibility of crossing species barrier, which is a feature of emerging viruses. Among the mammalian hosts, rodents are generally acknowledged to be important reservoirs of emerging viruses. Here, rodent samples collected from six provinces and autonomous regions of China (Liaoning, Inner Mongolia, Tibet, Xinjiang, Guangxi and Yunnan) were used to investigate the prevalence and distribution of bocaparvoviruses. By using next-generation sequencing first, a partial non-structural protein 1 (NS1) gene belonging to a possible novel bocaparvovirus was discovered. Following this, PCR-based screening of NS1 gene was conducted in 485 rodent samples, with 106 positive results found in seven rodent species (Rattus norvegicus, Mus musculus, Apodemus agrarius, Cricetulus barabensis, Rattus flavipectus, Rattus rattus and Rhombomys opimus). Finally, six nearly full-length genomes and three complete CDS were obtained and the newly identified bocaparvovirus was tentatively named rodent bocavirus (RoBoV). RoBoV has three ORFs: NS1, NP1, and VP, which are characteristics of bocaparvoviruses. Phylogenetic analyses revealed that porcine bocavirus isolate PBoV-KU14, a member of Ungulate bocaparvovirus 4, was the most related virus to RoBoV, with 92.1–92.9% amino acid identities in NS1 protein. Alignments of RoBoV-related sequences showed RoBoV isolates could be classified into two clades, demonstrating an inter-host genetic diversity. The results indicate a potential interspecies transmission of RoBoV between rodents and swine and expand our knowledge on bocaparvoviruses in rodent populations.
We assessed the practicality and effectiveness of using variation in the mitochondrial COI and COII genes to discriminate species and reconstruct the phylogeny of anophelene mosquitoes. Phylogenetic relationships among the subfamily Anophelinae were inferred from portions of the mitochondrial COI (92 species) and COII genes (108 species). Phylogenetic trees were reconstructed on the basis of parsimony, maximum likelihood and Bayesian methods. The suitability of COI and COII gene variation for identifying cryptic species was compared by comparing the sequence divergence within species groups and complexes. The results show that the COI gene was more useful for identifying sibling and cryptic species, but that phylogenetic relationships reconstructed using the COII gene were more similar to those based on morphological data. We conclude that: (1) there is a significant molecular divergence among An. sinensis; (2) the COI and COII are valid genetic markers for resolving taxonomic relationships among anopheline mosquitoes and the resultant phylogeny raises some questions about the taxonomic status of anopheline species groups and complexes; (3) the genus Anopheles is not demonstrably monophyletic with regard to the genus Bironella; (4) the subgenera Kerteszia and Nyssorhynchus are monophyletic; (5) below the group-level, COI data support the existence of monophyletic taxa within the Anopheles funestus, Anopheles maculipennis and Anopheles strode and Anopheles barbirostris subgroups, and within the Anopheles nuneztovari complex, whereas COII data support the monophyletic taxa within the Anopheles minimus and Anopheles oswaldoi subgroups, and Anopheles hyrcanus group. The monophyletic taxa within the Anopheles gambiae and Anopheles albitarsis complexes are supported by both COI and COII data.
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