Jinghui Zhao

Isolation of Irkut Virus from a Murina leucogaster Bat in China

Background and objectives Bats are recognized as a major reservoir of lyssaviruses; however, no bat lyssavirus has been isolated in Asia except for Aravan and Khujand virus in Central Asia. All Chinese lyssavirus isolates in previous reports have been of species rabies virus, mainly from dogs. Following at least two recent bat-associated human rabies-like cases in northeast China, we have initiated a study of the prevalence of lyssaviruses in bats in Jilin province and their public health implications. A bat lyssavirus has been isolated and its pathogenicity in mice and genomic alignment have been determined. Results We report the first isolation of a bat lyssavirus in China, from the brain of a northeastern bat, Murina leucogaster. Its nucleoprotein gene shared 92.4%/98.9% (nucleotide) and 92.2%/98.8% (amino acid) identity with the two known Irkut virus isolates from Russia, and was designated IRKV-THChina12. Following intracranial and intramuscular injection, IRKV-THChina12 produced rabies-like symptoms in adult mice with a short inoculation period and high mortality. Nucleotide sequence analysis showed that IRKV-THChina12 has the same genomic organization as other lyssaviruses and its isolation provides an independent origin for the species IRKV. Conclusions We have identified the existence of a bat lyssavirus in a common Chinese bat species. Its high pathogenicity in adult mice suggests that public warnings and medical education regarding bat bites in China should be increased, and that surveillance be extended to provide a better understanding of Irkut virus ecology and its significance for public health.

Ferret badger rabies origin and its revisited importance as potential source of rabies transmission in Southeast China

BackgroundThe frequent occurrence of ferret badger-associated human rabies cases in southeast China highlights the lack of laboratory-based surveillance and urges revisiting the potential importance of this animal in rabies transmission. To determine if the ferret badgers actually contribute to human and dog rabies cases, and the possible origin of the ferret badger-associated rabies in the region, an active rabies survey was conducted to determine the frequency of rabies infection and seroprevalence in dogs and ferret badgers.MethodsA retrospective survey on rabies epidemics was performed in Zhejiang, Jiangxi and Anhui provinces in southeast China. The brain tissues from ferret badgers and dogs were assayed by fluorescent antibody test. Rabies virus was isolated and sequenced for phylogenetic analysis. The sera from ferret badgers and dogs were titrated using rabies virus neutralizing antibodies (VNA) test.ResultsThe ferret badgers presented a higher percentage of rabies seroconversion than dogs did in the endemic region, reaching a maximum of 95% in the collected samples. Nine ferret badger-associated rabies viruses were isolated, sequenced, and were phylogenetically clustered as a separate group. Nucleotide sequence revealed 99.4-99.8% homology within the ferret badger isolates, and 83-89% homology to the dog isolates in the nucleoprotein and glycoprotein genes in the same rabies endemic regions.ConclusionsOur data suggest ferret badger-associated rabies has likely formed as an independent enzootic originating from dogs during the long-term rabies infestation in southeast China. The eventual role of FB rabies in public health remains unclear. However, management of ferret badger bites, rabies awareness and control in the related regions should be an immediate need.

Characterization of a rabies virus isolate from a ferret badger (Melogale moschata) with unique molecular differences in glycoprotein antigenic site III

Rabies virus was isolated from the brain of a Chinese ferret badger (Melogale moschata) and identified as having an R333Q substitution within its glycoprotein antigenic site III. Additionally, compared with vaccine strains and other rabies virus isolates from dogs and ferret badgers in China, the isolate had five other amino acid substitutions in its glycoprotein: P(-17)L in the signal peptide, R88H, L225M, and D422E in the ectoplasmic region, and G478E in the cytoplasmic region. This isolate possessed high virulence in suckling, weanling and adult mice. These data indicate that this is a unique rabies virus with a molecular signature that differentiates it from other strains circulating in terrestrial mammals in China. We propose that rabies virus circulates in some ferret badgers in an independent epidemiological cycle unique to China following spillover from domestic dogs or other hosts infected with rabies.

Analysis of an Outbreak of Human Rabies in 2009 in Hanzhong District, Shaanxi Province, China

Between March and August 2009, there was an outbreak of rabies in both humans and dogs in Hanzhong District, Shaanxi province, China. About 7300 humans were bitten by dogs and 20 died of rabies due to failure to perform postexposure prophylaxis. The local authorities therefore conducted a dog slaughter campaign. From a random selection of brains of dogs culled in the campaign, 0/27 tested positive for rabies virus by immunofluorescence. Of two dogs known to have bitten humans, one was shown to contain live rabies virus by immunofluorescence and mouse intracerebral inoculation. Serological studies during the outbreak revealed that only 1/27 dog was antibody positive: after a mass vaccination campaign, 20.8% seroconverted. Lack of canine vaccination was clearly the main reason for dog rabies spread and human infection. Phylogenetic analysis of a virus isolate showed that its genomic sequence was closely related to the clade 1 rabies strains widely circulating in China. The highest homology was found with the isolate circulating in Sichuan province, a neighboring province south of Shaanxi, indicating the spread of rabies from the south to the north.

Molecular characterization of a rabies virus isolate from a rabid dog in Hanzhong District, Shaanxi Province, China.

A canine rabies virus, Shaanxi-HZ-6, was isolated in Shaanxi Province, China, in 2009. Its genome has been completely sequenced and found to be closely related to the China I rabies virus strains widely circulating in China. The genomic length was 11,923 base pairs, and the overall organization of the genome was similar to that of other rabies virus isolates. Compared with isolates CQ92 and J, 84 amino acid substitutions (7 in the N gene, 15 in P, 6 in M, 25 in G, 31 in L) were observed in strain Shaanxi-HZ-6. Amino acid substitutions of R264H and V332I were noted in the G protein antigenic site I and site III, respectively. Residue 333 of the G protein, which is considered to be associated with pathogenicity, was Arg in Shaanxi-HZ-6. These and other substitutions may help provide an explanation why the China I lineage strain maintains its prevalence in China.

Complete Genome Sequence of a Rabies Virus Isolate from a Ferret Badger (Melogale moschata) in Jiangxi, China

ABSTRACT The genome of ferret badger rabies virus JX09-17(fb), isolated in Jiangxi Province, China, in 2009 has been completely sequenced. The genomic length is 11, 923 nucleotides (nt) with an overall organization similar to that of other rabies virus isolates. JX09-17(fb) is closely related to Chinese epidemic canine isolates in clade I.

Fox- and raccoon-dog-associated rabies outbreaks in northern China

Dear Editor,Rabies is a generally fatal disease caused by the rabies virus(RABV),and is transmitted mainly by Carnivora and Chiroptera(Fooks A R,et al.,2014;Tao X,et al.,2013).In China,stray dogs and some wild animals(e.g.,Chinese ferret badgers,foxes,and raccoon dogs)are the principal reservoirs for RABV(Hu R L,et al.,2009).Historically,rabies in wild foxes and raccoon dogs(Nyctereutes procyonoides)was recorded in the early

Experimental Oral Immunization of Ferret Badgers (Melogale moschata) with a Recombinant Canine Adenovirus Vaccine CAV-2-E3Δ-RGP and an Attenuated Rabies Virus SRV9

Abstract Ferret badgers (Melogale moschata) are a major reservoir of rabies virus in southeastern China. Oral immunization has been shown to be a practical method for wildlife rabies management in Europe and North America. Two groups of 20 ferret badgers were given a single oral dose of a recombinant canine adenovirus-rabies vaccine, CAV-2-E3&Dgr;-RGP, or an experimental attenuated rabies virus vaccine, SRV9. At 21 days, all ferret badgers had seroconverted, with serum virus-neutralizing antibodies ranging from 0.1 to 4.5 IU/mL. Titers were >0.50 IU/mL (an acceptable level) in 17/20 and 16/20 animals receiving CAV-2-E3&Dgr;-RGP or SRV9, respectively. The serologic results indicate that the recombinant CAV-2-E3&Dgr;-RGP is at least as effective as the attenuated rabies virus vaccine. Both may be considered for additional research as oral rabies vaccine candidates for ferret badgers.

Epidemic and Maintenance of Rabies in Chinese Ferret Badgers (Melogale moschata) indicated by Epidemiology and the Molecular Signatures of Rabies Viruses

An epidemic of Chinese ferret badger-associated human rabies was investigated in Wuyuan county, Jiangxi province and rabies viruses isolates from ferret badgers in different districts in Jiangxi and Zhejiang provinces were sequenced with their nucleotides and amino acids and aligned for epidemiological analysis. The results showed that the human rabies in Wuyuan are only associated with ferret badger bites; the rabies virus can be isolated in a high percentage of ferret badgers in the epidemic areas in Jiangxi and Zhejiang provinces; the isolates share the same molecular features in nucleotides and have characteristic amino acid signatures, i.e., 2 sites in the nucleoprotein and 3 sites in the glycoprotein, that are distinct from virus isolates from dogs in the same region. We conclude that rabies in Chinese ferret badgers has formed an independent transmission cycle and ferret badgers may serve as another important rabies reservoir independent of dog rabies in China.

A SYBR-green I quantitative real-time reverse transcription-PCR assay for rabies viruses with different virulence.

Dear Editor,To establish a more specific diagnostic assay than thecurrent methods for rabies virus(RABV)infection in ca-nines and wildlife species,we developed a SYBR-GreenI quantitative real-time reverse transcription-PCR(RT-PCR).Three primers were designed and synthesized:oneof which,located within the leader sequences and target-ed for the whole viral genome amplification to obtain its