We report the isolation and characterization of a novel bat coronavirus which is much closer to the severe acute respiratory syndrome coronavirus (SARS-CoV) in genomic sequence than others previously reported, particularly in its S gene. Cell entry and susceptibility studies indicated that this virus can use ACE2 as a receptor and infect animal and human cell lines. Our results provide further evidence of the bat origin of the SARS-CoV and highlight the likelihood of future bat coronavirus emergence in humans.T he 2002-2003 outbreak of severe acute respiratory syndrome coronavirus (SARS-CoV) was a significant public health threat at the beginning of the 21st century (1). Initial evidences showed that the masked palm civet (Paguma larvata) was the primary suspect in the animal origin of SARS-CoV (2, 3). Later studies suggested that Chinese horseshoe bats are natural reservoirs and that the masked palm civet most likely served as an intermediate amplification host for SARS-CoV (4, 5). From our longitudinal surveillance of bat SARS-like coronavirus (SL-CoV) in a single bat colony of the species Rhinolophus sinicus in Kunming, Yunnan Province, China, we found a high prevalence of diverse SL-CoVs (6). Whole-genome sequence comparison revealed that these SL-CoVs have 78% to 95% nucleotide sequence identities to SARS-CoV, with the major differences located in the spike protein (S) genes and the region of open reading frame 8 (ORF8). We recently isolated a bat SL-CoV strain (WIV1) and constructed an infectious clone of another strain (SHC014); significantly, these strains are closely related to SARS-CoV and capable of using the same cellular receptor (angiotensin-converting enzyme 2 [ACE2]) as SARS-CoV (6, 7). Despite the high similarity in genomic sequences and receptor usage of these two strains, there is still some difference between the N-terminal domains of the S proteins of SARS-CoV and other SL-CoVs, indicating that other unknown SL-CoVs are circulating in bats.Here we report the isolation of a new SL-CoV strain, named bat SL-CoV WIV16. SL-CoV WIV16 was isolated from a single fecal sample of Rhinolophus sinicus, which was collected in Kunming, Yunnan Province, in July 2013. The full genomic sequence of SLCoV WIV16 (GenBank accession number KT444582) was determined and contained 30,290 nucleotides (nt) and a poly(A) tail which is slightly larger than those of SARS-CoVs and other bat SL-CoVs (6, 8-13). The WIV16 genome has a 40.9% GϩC content and short untranslated regions (UTRs) of 264 and 339 nt at the 5= and 3= termini, respectively. Its gene organization is identical to that of WIV1 and slightly different from that of the civet SARS-CoV and other bat SL-CoVs due to an additional ORF (name ORFx) detected between the ORF6 and ORF7 genes of the WIV1 and WIV16 genomes (data not shown). The conserved transcriptional regulatory sequence was identified upstream of ORFx, indicating that this is likely to be a potential functional gene. The overall nucleotide sequence of WIV16 has 96% identity (higher than that of any p...
Self-healing materials are notable for their ability to recover from physical or chemical damage. We report that commodity copolymers, such as poly(methyl methacrylate)/n-butyl acrylate [p(MMA/nBA)] and their derivatives, can self-heal upon mechanical damage. This behavior occurs in a narrow compositional range for copolymer topologies that are preferentially alternating with a random component (alternating/random) and is attributed to favorable interchain van der Waals forces forming key-and-lock interchain junctions. The use of van der Waals forces instead of supramolecular or covalent rebonding or encapsulated reactants eliminates chemical and physical alterations and enables multiple recovery upon mechanical damage without external intervention. Unlike other self-healing approaches, perturbation of ubiquitous van der Waals forces upon mechanical damage is energetically unfavorable for interdigitated alternating/random copolymer motifs that facilitate self-healing under ambient conditions.
bIncreasing data indicate that bats harbor diverse viruses, some of which cause severe human diseases. In this study, sequenceindependent amplification and high-throughput sequencing (Solexa) were applied to the metagenomic analysis of viruses in bat fecal samples collected from 6 locations in China. A total of 8,746,417 reads with a length of 306,124,595 bp were obtained. Among these reads, 13,541 (0.15%) had similarity to phage sequences and 9,170 (0.1%) had similarity to eukaryotic virus sequences. A total of 129 assembled contigs (>100 nucleotides) were constructed and compared with GenBank: 32 contigs were related to phages, and 97 were related to eukaryotic viruses. The most frequent reads and contigs related to eukaryotic viruses were homologous to densoviruses, dicistroviruses, coronaviruses, parvoviruses, and tobamoviruses, a range that includes viruses from invertebrates, vertebrates, and plants. Most of the contigs had low identities to known viral genomic or protein sequences, suggesting that a large number of novel and genetically diverse insect viruses as well as putative mammalian viruses are transmitted by bats in China. This study provides the first preliminary understanding of the virome of some bat populations in China, which may guide the discovery and isolation of novel viruses in the future. Bats, the only flying mammal and the second most diverse group of mammals, are natural reservoirs of many emerging viruses (7, 38). Some bat viruses cause severe human diseases, such as lyssavirus, Hendra virus, Nipah virus, Ebola virus, and Marburg virus (16,23,29,41,47). Most of the known bat viruses have been discovered in apparently healthy bats (10,14,19,20,26,34,49). When bats are experimentally infected with henipavirus or the rabies virus, the bats shed the virus but do not produce any clinical syndrome like those observed in other animals and humans (1,17,30,40). This phenomenon may be due to the adaption of viruses to their host species, preinfection with related nonpathogenic viruses, or some unique characteristics of the bat immune system to viral infection. Considering the species richness and wide geographic distribution of bats, bats may be a source of many additional unknown viruses.The techniques for isolating viruses are straightforward but are not always successful because the majority of viruses are unable to grow in cell culture due to a lack of susceptible cell lines, a low titer of virus, or toxicity of environmental samples. PCR assays are sequence-dependent and extremely sensitive and rapid but have been complicated by the lack of a universally conserved gene or gene marker for all types of viruses. Recently, the sequence-independent amplification of viral nucleic acids followed by shotgun sequencing or pyrosequencing has been used to discover an enormous diversity of virus species and genotypes in marine and fresh water (2,6,12,37,39,45), human feces (3, 5, 43), animal tissues, and bat feces (11,13,24,32,42).In this study, six bat fecal samples were collected from six locations...
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