Quan Shen

Hepatitis E Virus Infection among Domestic Animals in Eastern China

Hepatitis E virus (HEV) is a zoonotic pathogen of which several species of animal were reported as reservoirs. Antibodies to HEV and HEV RNA have been detected in some Chinese population and swine groups but few other domestic animals. In this study, to investigate the HEV prevalence, we tested sera from 788 pigs, 100 cows, 50 goats, 49 horses, 101 pet dogs, 105 chickens, 47 duck and 45 pigeons in eastern China for anti‐HEV immunoglobulin G (IgG). We also tested 50% of the swine sera, all of sera from the other domestic animals and 13 Shanghai human sera which were positive for anti‐HEV immunoglobulin M (IgM) for HEV RNA using reverse transcriptase‐polymerase chain reaction. Our results indicated that 82.5% (222/269) of the sows, 53.9% (104/193) of the 4‐ to 6‐month‐old swine, 63.4% (168/265) of the 1‐ to 3‐month‐old swine, 55.7% (34/61) of the slaughterhouse swine, 24% (12/50) of the goats, 16.3% (8/49) of the horses, 17.8% (21/101) of the pet dogs, 6% (6/100) of the cows, 12.8% (6/47) of the ducks, 4.4% (2/45) of the pigeons and 1.9% (2/105) of the chickens exhibited positive for anti‐HEV IgG. Inhibition assay confirmed the infection with HEV or HEV‐like viruses in these domestic animals except pigeons and chickens. From the sera, we isolated 18 swine HEV strains, one horse HEV strain and two human HEV strains. Sequence analysis showed that the horse HEV isolate and one swine isolate belonged to genotype 3. The other isolates belonged to genotype 4. The two human isolates were phylogenetically closely related to eight of the swine isolates. In short, the presence of anti‐HEV antibody had been confirmed in several species of domestic animals in eastern China and HEV RNA has been identified in swine, human and horse. This suggested that the authorities should pay more attention to the prevalence of HEV in eastern China.

Cross-species infection of hepatitis E virus in a zoo-like location, including birds

Hepatitis E virus (HEV) is a zoonotic pathogen of which several species of animals are considered to be reservoirs. Thirty-eight faecal samples, obtained from 22 species of animals including birds in a wildlife first-aid centre in Eastern China, were tested for HEV RNA. Our survey revealed that in total 28.9% (95% confidence interval 14.5-43.4) of the faecal samples from various mammals and birds were HEV RNA positive. Sequence and phylogenetic analyses of the 11 isolates demonstrated that all sequences clustered in genotype 4 with 96-100% identity to each other. In addition, serum samples from seven animal handlers have shown that five (71.4%) were seropositive. The findings imply that cross-species infection of HEV had probably occurred in this zoo-like location, and moreover, birds can be infected naturally with mammalian HEV.

Hepatitis E Virus Infection in Central China Reveals No Evidence of Cross-Species Transmission between Human and Swine in This Area

Hepatitis E virus (HEV) is a zoonotic pathogen of which several species of animal were reported as reservoirs. Swine stands out as the major reservoir for HEV infection in humans, as suggested by the close genetic relationship of swine and human virus. Since 2000, Genotype 4 HEV has become the dominant cause of hepatitis E disease in China. Recent reports showed that genotype 4 HEV is freely transmitted between humans and swine in eastern and southern China. However, the infection status of HEV in human and swine populations in central China is still unclear. This study was conducted in a rural area of central China, where there are many commercial swine farms. A total of 1476 serum and 554 fecal specimens were collected from the general human and swine populations in this area, respectively. The seroepidemiological study was conducted by enzyme-linked immunosorbent assay. Conserved genomic sequences of open reading frame 2 were detected using reverse transcription-PCR. The results indicated that the overall viral burden of the general human subjects was 0.95% (14/1476), while 7.0% (39/554) of the swine excreted HEV in stool. The positive rate of anti-HEV IgG and IgM in the serum samples was 7.9% (117/1476) and 1.6% (24/1476), respectively. Phylogenetic analysis based on the 150 nt partial sequence of the capsid protein gene showed that the 53 swine and human HEV isolates in the current study all belonged to genotype 4, clustering into three major groups. However, the HEV isolates prevalent in the human and swine populations were classified into known distinct subgenotypes, which suggested that no cross-species transmission between swine and humans had taken place in this area. This result was confirmed by cloning and phylogenetic analysis of the complete capsid protein gene sequence of three representative HEV strains in the three major groups. The cross reactivity between anti-HEV IgG from human sera and the two representative strains from swine in central China was confirmed by Dot-blot assay. In conclusion, although all the HEV strains prevalent in central China belonged to genotype 4, there is no evidence of cross-species transmission between human and swine in this area.

Prevalence of antibody to hepatitis E virus among pet dogs in the Jiang-Zhe area of China

In order to investigate the prevalence of HEV infection and immunity in pet dogs in the Jiang-Zhe area of China, we examined 192 dog serum samples collected from 11 animal hospitals in this area for anti-HEV IgG and HEV RNA, using ELISA and RT-PCR assays, respectively. Two dogs were injected intravenously with swine HEV to obtain anti-HEV antibody positive serum for ELISA assay. After the ELISA assay, 26 samples showing OD values higher than 0.20 were selected and examined by Dot-blot assay, and 88.5% (23/26) were confirmed to be positive for anti-HEV IgG. Although our results indicated swine HEV could experimentally induce anti-HEV IgG in the serum, we failed to detect HEV RNA in the serum samples from either the 2 intravenously injected dogs or 192 serum samples collected from animal hospitals. This implied that dog may not be a reservoir of HEV in this area.

The First Chinese Porcine Sapovirus Strain That Contributed to an Outbreak of Gastroenteritis in Piglets

Sapovirus (SaV) belongs to the family Caliciviridae and is a causative agent of gastroenteritis in children and adults ([6][1], [2][2]). On the basis of complete capsid gene sequences, SaV can be divided into five genogroups, among which GI, GII, GIV, and GV infect humans, whereas SaV GIII infects

Prevalence of four different subgenotypes of genotype 4 hepatitis E virus among swine in the Shanghai area of China

BackgroundHepatitis E virus (HEV) is a zoonotic pathogen of which swine was reported as major reservoirs. HEV has been divided into 4 different genotypes according to phylogenetic analysis. Recent reports showed that genotype 4 HEV is freely transmitted between humans and swine in eastern China, including Shanghai area. This paper investigated the recent infection status of HEV among swine population of Shanghai area in China.Methods480 swine faecal specimens were collected from 23 farms which distribute all over Shanghai from September to November, 2007 and tested for the presence of HEV RNA by the polymerase chain reaction (PCR).ResultsOur results showed that 26.1% (6/23) of the swine farms were positive for HEV RNA and the positive rate of the six farms were ranged from 9.1% to 33.3%. The HEV RNA positive rate for total samples were 5% (24/480). The resulted positive band specific for HEV was sequenced and sequence analysis indicated that all of these isolates belonged to genotype 4 HEV. Phylogenetic analysis showed that the 24 isolates clustered into 4 distinct subgroups, sharing 83.3–89.7% inter-subgroup and 97–99% intra-subgroup identities. More over, isolates in three of the four subgroups closely clustered with previous identified strains, sharing up high to 97% identity with them.ConclusionThese results suggested that there were 4 different subgenotypes of HEV prevalent in Shanghai, and some of them may not be indigenous to Shanghai but introduced from other geographic regions.

MOLECULAR DETECTION AND PREVALENCE OF PORCINE CALICIVIRUSES IN EASTERN CHINA FROM 2008 TO 2009

Caliciviruses causing diarrhea have been reported in both industrial and developing countries, including China, in recent years. Porcine caliciviruses that are closely related to human sapoviruses (SaVs) and noroviruses (NoVs) have also been detected in swine, which has raised discussion about the animal reservoir and the potential risk for zoonotic transmission to humans. The objective of this work was to determine the frequency and age distribution of SaVs and NoVs in pigs and to characterize the strains prevalent in eastern China. A total of 904 stool samples from pigs of different ages were collected from eastern China from April 2008 to March 2009 and tested for both SaVs and NoVs using reverse transcription-polymerase chain reaction (RT–PCR). Our results indicate that 8 (0.9%) stool samples were positive for SaVs and 2 (0.2%) for NoVs. Phylogenetic analysis of partial sequences of the RNA-dependent RNA polymerase (RdRp) gene indicated that all of the SaV strains belonged to the GIII SaVs, while the two NoV strains belonged to the GII NoV genogroup. The 8 SaV strains were further divided into two clusters, which clustered closely with the Netherlands isolate (AY615804) and the Chinese strain (EU599212), respectively. The two NoV strains shared about 67.3–67.6% nucleotide homology with a human norovirus strain (DQ369797), the only NoV strain from mainland China available in GenBank. Moreover, our results suggest that SaV infections are more frequent in 0-1xa0month-old pigs than in older ones. In conclusion, the present study provides evidence that PoSaVs and PoNoVs exist in swine in eastern China.

Genomic organization and recombination analysis of human norovirus identified from China

Noroviruses (NoVs) are one of the major causal agents of acute gastroenteritis in both industrial and developing countries including China. Recent studies have revealed that NoV genome is highly prone to mutation and recombination which may lead to emergence of new strains. In the present study, three full-length genomes of human NoV from China were determined and the genomic organization and recombination were analyzed. They had similar genome organization and contained three predicted ORFs, though the 5′UTR of those three strains were 2, 4 and 8 nucleotides, respectively. Phylogenetic analysis showed that the HU/GII/SHANGHAI/SH312/2008/CHN strain may be a recombinant of GII-3 capsid and GII-4 polymerase. To confirm the finding and detect the breakpoints where the recombination event occurred, we performed recombination analysis based on the genomic sequences of HU/GII/SHANGHAI/SH312/2008/CHN as the query sequence, and AB220921/NOV/JP/GII-4 and AB365435/NOV/US/GII-3 as the background sequences, using RPD software. Results indicated that the two parental strains were AB220921/NOV/JP/GII-4 and AB365435/NOV/US/GII-3. The breakpoint for this recombination event located at position 5,107xa0nt of the genome (in the ORF1 and ORF2 overlap).

Isolation and characterization of a genotype 4 Hepatitis E virus strain from an infant in China

In the present study, a genotype 4 HEV strain was identified in the fecal specimen from a seven months old infant with no symptom of hepatitis in Shanghai Childrens hospital. The full capsid protein gene (ORF2) sequence of this strain was determined by RT-PCR method. Sequence analysis based on the full ORF2 sequence indicated that this HEV strain shared the highest sequence identity (97.6%) with another human HEV strain isolated from a Japanese patient who was infected by genotype 4 HEV during traveling in Shanghai. Phylogenetic analysis showed that this genotype 4 HEV was phylogenetically far from the genotype 4 HEV strain that was commonly prevalent in Shanghai swine group, suggesting that this strain may not come from swine group and not involved in zoonotic transmission in this area.

Molecular characterization and phylogenetic analysis of the complete genome of a porcine sapovirus from Chinese swine

BackgroundPorcine sapovirus was first identified in the United States in 1980, hitherto, several Asian countries have detected this virus. In 2008, the first outbreak of gastroenteritis in piglets caused by porcine sapovirus in China was reported. The complete genome of the identified SaV strain Ch-sw-sav1 was sequenced and analyzed to provide gene profile for this outbreak.MethodsThe whole genome of Ch-sw-sav1 was amplified by RT-PCR and was sequenced. Sequence alignment of the complete genome or RNA dependent RNA polymerase (RdRp) gene was done. 3 end of ORF2 with 21-nt nucleotide insertion was further analyzed using software.ResultsSequence analysis indicated that the genome of Ch-sw-sav1 was 7541 nucleotide long with two ORFs, excluding the 17 nucleotides ploy (A) at the 3 end. Phylogenetic analysis based on part of RdRp gene of this strain showed that it was classified into subgroup GIII. Sequence alignment indicated that there was an inserted 21-nt long nucleotide sequence at the 3 end of ORF2. The insertion showed high antigenicity index comparing to other regions in ORF2.ConclusionCh-sw-sav1 shared similar genetic profile with an American PEC strain except the 21-nt nucleotide at the 3 end of ORF2. The insert sequence shared high identity with part gene of Sus scrofa clone RP44-484M10.