东北边境地区野猪及放养杂交野猪戊型肝炎流行病学调查

为了解东北边境地区野猪及放养杂交野猪群体猪戊型肝炎病毒(HEV)感染情况,于2015—2018年在吉林省、黑龙江省的中朝、中俄边境和内蒙古自治区境内加格达奇周边地区采集6月龄以上杂交野猪血清、粪便或肛拭子样品共520份,采集野猪血清和粪便样品共248份。ELISA检测、RT-nPCR检测、全基因组测序、同源性及进化分析结果显示,杂交野猪和野猪感染HEV的血清抗体总阳性率为34.1%(136/399);核酸总阳性率为1.56%(12/771),12份核酸阳性样品均来自杂交野猪,病毒基因组ORF2部分核苷酸序列同源性为85.4%～100.0%,属于基因4型,4a、4b亚型。对4a亚型的1份阳性样品(LJG-18)进行病毒全基因组扩增测序,其核苷酸序列与日本的人源毒株JKO-ChiSai98C同源性最高,为94.9%,与吉林省猪源毒株Ch-S-1同源性为90.2%。结果表明:东北边境地区放养杂交野猪群具有较高的HEV血清抗体阳性率,HEV流行毒株以4a亚型为主。本试验针对我国野猪及放养杂交野猪群体开展猪戊型肝炎流行病学调查,为该病的流行情况提供了新的科学数据,对我国养猪业健康发展和公共卫生安全具有重要意义。     

Nationwide survey of hepatitis E virus infection among wildlife in Japan

In Japan, hepatitis E virus (HEV) causes hepatitis in humans through the consumption of raw or undercooked meat, including game meat. In the present study, nationwide surveillance of HEV infection among a total of 5,557 wild animals, including 15 species, was conducted in Japan. The prevalence of anti-HEV antibodies in wild boar was 12.4%, with higher positive rates in big boars (over 50 kg, 18.4%) than in small individuals (less than 30 kg, 5.3%). Furthermore, HEV RNA was more frequently detected in piglets than in older boars. Interestingly, the detection of HEV among wildlife by ELISA and RT-PCR suggested that HEV infection in Sika deer was a very rare event, and that there was no HEV infection among wild animals except for wild boar, Sika deer and Japanese monkeys. In conclusion, wild boar, especially piglets, are at high risk of HEV infection, while other wild animals showed less risk or no risk of HEV transmission.     

Serological survey of antibodies to four pathogens in wild boars in Nagano Prefecture,Japan

Wild boars (Sus scrofa) captured or found dead in Nagano Prefecture were surveyed for antibodies to Aujeszky’s disease virus (ADV), Erysipelothrix rhusiopathiae, porcine reproductive and respiratory syndrome virus (PRRSV), and Toxoplasma gondii. While all 168 samples tested were negative for anti-ADV antibodies and all 140 samples tested were negative for anti-PRRSV antibodies, all 190 samples tested were positive for anti-E. rhusiopathiae antibodies and 12 of 180 samples were positive for anti-toxoplasma antibodies. These results suggest that since E. rhusiopathiae and T. gondii cause zoonotic diseases, in addition to wild boars being a potential source of infection for domestic pigs, caution should be taken when handling wild boars or eating wild boar meat because of the possibility of human infection.     

Serological evidence for influenza virus infection in Korean wild boars

Serum samples from 1,011 wild boars hunted in 2012 were collected for serological surveillance for 4 subtypes (pandemic A (H1N1) 2009 and classical H1N1, H1N2 and H3N2) of swine influenza virus (SIV). Samples from 12 of the boars were identified as positive for SIV (pandemic A (H1N1) 2009, n=9; classical H1N1, n=2; and H1N2, n=1) by a hemagglutination inhibition test (HI test) and a nucleoprotein (NP)-based enzyme-linked immunosorbent assay (NP-ELISA). Although the overall seroprevalence of SIV in the Korean wild boar population was somewhat low compared with that in China and the U.S.A., the apparent prevalence of pandemic H1N1 was notable. Therefore, continuous monitoring of the wild boar population is needed as it may be a major reservoir for pandemic H1N1, facilitating its spread to humans and domestic pigs.     

猪戊型肝炎病毒荧光定量PCR检测方法的建立及其分子流行病学

对已发表的戊型肝炎病毒(HEV)不同基因型毒株全序列进行分析,针对ORF3的保守区设计引物和探针优化建立了HEV荧光RT-PCR检测体系。分析表明该反应体系具有良好的扩增效率、特异性和稳定性,且检测灵敏度比普通RT-PCR方法高10～100倍。利用所建立的荧光定量PCR方法对采集自浙江及其周边上海、江苏等地区规模化猪场的样品以及HEV抗体阳性人血清样品进行了HEV核酸检测。其中,77份人血清样本仅有2份为HEV核酸阳性(2.6%)。而猪粪便样品阳性率则高达20.5%(56/273),且所调查的猪场均存在HEV感染。进一步的分子流行特征分析表明,浙江及周边地区的猪HEV毒株多为基因Ⅳ型,与其他地区的Ⅳ型HEV毒株同源性较高(83.4%～89.5%),但它们之间也存在较多的变异位点。猪源HEV毒株PJ-1则与多数Ⅲ型HEV毒株同源性较高(88%),进化分析也表明其为基因Ⅲ型。人血清样品中检测到的2份HEV毒株Human-1与Human-2与本地区Ⅳ型猪源毒株在分子进化关系上具有很高的亲源性,它们分布于同一分支内,而不构成独立分支。以上分析结果表明浙江及其周边地区猪HEV感染较为广泛,且以基因Ⅳ型毒株为主,但也存在...     

A serological survey of porcine reproductive and respiratory syndrome virus in wild boar in Gifu Prefecture,Japan

Porcine reproductive and respiratory syndrome (PRRS) is an infectious swine disease caused by the PRRS virus (PRRSV) that results in economic loss to the pig-rearing industry. To study PRRSV infection in wild boars and pigs, we conducted a serological survey in Gifu Prefecture, Japan, from 2020 to 2021. Three out of 453 (0.7%) wild boar sera were positive for PRRSV antibodies in a commercial ELISA. However, given that PRRSV RNA was not detected in these three wild boars and the specificity and sensitivity of the test kit, these are considered as false positives. Although seropositive pigs were found in multiple pig farms in the study area, the role of wild boars as a source of PRRS to pig farms appeared to be minimal.     

Detection of hepatitis E virus RNA in saliva for diagnosis of acute infection

Diagnosis of acute hepatitis E virus (HEV) infection is established by detection of anti‐HEV IgM antibodies by ELISA or by amplification of serum viral RNA. Here, we evaluate the diagnostic value of testing HEV RNA in saliva to identify patients with acute HEV infection. Prospective proof‐of‐concept study including patients with acute hepatitis. Whole blood and neat saliva samples were obtained from all patients. Saliva samples were processed and analysed for HEV RNA by RT‐PCR within 2 hr after collection. A total of 34 patients with acute hepatitis and 12 healthy donors were included in the study. HEV RNA in serum was confirmed by RT‐PCR in eight of these patients (23.5%; 95% CI: 12.2%–40.2%). HEV was isolated in the saliva of eight of 34 patients (23.5%; 95% CI: 12.2%–40.2%). All patients with HEV RNA amplified in saliva had detectable HEV RNA in serum. HEV was isolated neither in the saliva of any of the 26 patients without detectable HEV RNA in serum nor in healthy donors. Our study suggests that acute HEV infection could be diagnosed by assessing viral load in saliva.     

Development of a novel enzyme-linked immunosorbent assay to detect anti-IgG against swine hepatitis E virus

Swine hepatitis E virus (HEV) is widespread throughout pigs in both developing and industrialized countries. This virus is an important zoonotic agent and a public concern worldwide. Infected pigs are asymptomatic, so diagnosing swine HEV relies on detection of the virus or antibodies against the virus. However, several obstacles need to be overcome for effective and practical serological diagnosis. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) that used a purified recombinant capsid protein of swine HEV. The potential clinical use of this assay was evaluated by comparing it with a commercial kit (Genelabs Technologies, Diagnostics, Singapore). Results of the ELISA were highly correlated with those of the commercial kit with a sensitivity of 97% and specificity of 95%. ROC (receiving operator characteristic) analysis of the ELISA data produced a value of 0.987 (95% CI, 0.977~0.998, p < 0.01). The cut-off value for the ELISA was also determined using negative pig sera. In summary, the HEV-specific ELISA developed in the present study appears to be both practical and economical.     

Endemic infections of bovine viral diarrhea virus genotypes 1b and 2a isolated from cattle in Japan between 2014 and 2020

Bovine viral diarrhea virus (BVDV) is a causative agent of bovine viral diarrhea. In Japan, a previous study reported that subgenotype 1b viruses were predominant until 2014. Because there is little information regarding the recent epidemiological status of BVDV circulating in Japan, we performed genetic characterization of 909 BVDV isolates obtained between 2014 and 2020. We found that 657 and 252 isolates were classified as BVDV-1 and BVDV-2, respectively, and that they were further subdivided into 1a (35 isolates, 3.9%), 1b (588, 64.7%), 1c (34, 3.7%), and 2a (252, 27.7%). Phylogenetic analysis using entire E2 coding sequence revealed that a major domestic cluster in Japan among BVDV-1b and 2a viruses were unchanged from a previous study conducted from 2006 to 2014. These results provide updated information concerning the epidemic strain of BVDV in Japan, which would be helpful for appropriate vaccine selection.     

Seroprevalence of hepatitis E virus in dogs in Switzerland

Hepatitis E virus (HEV) is the causative agent of an acute and in most cases self‐limiting hepatitis. Of the four major HEV genotypes that infect humans, genotype 3 and 4 are zoonotic and have been identified in humans but predominantly in pigs and wild boar, which are considered the main reservoirs. However, the known host range of zoonotic HEV may be increasing to comprise additional species, including companion animals. Several studies have identified contact with dogs as a risk factor for HEV infection in humans, yet information on the occurrence of HEV in Swiss dogs is lacking. To examine a possible risk of exposure, this study was designed to assess the seroprevalence of HEV in 84 Swiss dogs. Serum and plasma samples collected from four veterinary clinics were screened for HEV‐specific antibodies by HEV‐antibody ELISA test kit. In addition, information of 22 dogs regarding the country of origin, the type of dog feed and any history of hunting was recorded. Samples from seropositive animals were also screened for the presence of HEV RNA by quantitative real‐time RT‐PCR (qRT‐PCR). Overall, 38% (32 of 84) of the dogs tested seropositive for anti‐HEV, indicating exposure to HEV. Among the 22 dogs for which information was available, HEV‐specific antibodies were detected in three of five dogs that were born abroad, in one of two dogs that were fed a raw meat‐based diet, and in one hunting dog. No viral RNA could be detected in any of the serum and plasma samples; thus, the genotype of the strains remained undetermined. This study provides further evidence for canine exposure and susceptibility to HEV and highlights the need to further assess the risks of HEV transmission to humans with contact to dogs.     

Prevalence of hepatitis E virus infection among people and swine in mainland China: A systematic review and meta‐analysis

Hepatitis E virus (HEV) infection remains an important public health problem, and it is endemic primarily in developing countries. This study aimed to evaluate the seroprevalence of HEV among the general population, occupational population and swine in mainland China and its risk factors based on a systematic review and meta‐analysis. Systematic search from EMBASE, PubMed, Web of Science, Cochrane Library and several Chinese databases, such as Wanfang (WF) Data, China National Knowledge Infrastructure (CNKI), Chinese Science and Technology Periodical Database (VIP) and SINOMED, was searched from inception up to 25 April 2018. The overall seroprevalence of HEV and its corresponding 95% confidence interval (CI) as well as the correlation coefficients between different groups were estimated using stata 12.0 and r ‐3.4.1 software. Potential sources of heterogeneity were explored using subgroup and sensitivity analyses. Twenty‐eight studies with 57,274 participants (including human and swine) were included. The seroprevalence of anti‐HEV immunoglobulin G (IgG) among the general population, occupational population and swine was 27.3% (95% CI: 22.4–32.2), 47.4% (95% CI: 40.1–54.8) and 66.4% (95% CI: 61.7–71.1), respectively. The overall prevalence of IgM among the general population was 1.8% (95% CI: 0.7–2.9). The odds ratio for the occupational population, as compared to the general population, was 2.63. The highest anti‐HEV IgG prevalence (59%) was observed in East China, whereas the lowest (34.8%) was noted in Northeast and North China. In the occupational population, the highest prevalence (77.0%) was observed among swine vendors. Seven studies included 30,392 participants (humans and swine); the correlation coefficient for the prevalence of anti‐HEV IgG between the professional population and adult pigs was 0.88. Sensitivity analyses showed that the stability of results was not considered significant. This research found that HEV is common in China, and contact with pork or other pig products may be an important mode of HEV transmission.     

Pseudorabies virus infection in hunting dogs in Oita,Japan: Report from a prefecture free from Aujeszky’s disease in domestic pigs

We isolated two pseudorabies virus (PRV) isolates (designated OT-1 and OT-2) from two hunting dogs exhibiting neurological manifestations after eating the flesh of wild boar hunted in Oita prefecture, Kyushu Island, Japan. The isolates corresponded to a previously reported PRV (MY-1 strain) isolated from a hunting dog in neighboring Miyazaki prefecture, and it clustered into genotype II based on the glycoprotein C sequence. Our results suggest that this common PRV strain may have been maintained in wild boars on Kyushu Island even though domestic pigs in this area have attained an Aujeszky’s disease-free status.     

Detection of Mycobacterium tuberculosis complex antibodies in free-ranged wild boar and wild macaques in selected districts in Selangor and reevaluation of tuberculosis serodetection in captive Asian elephants in Pahang,Peninsular Malaysia

Tuberculosis (TB) is a chronic inflammatory and zoonotic disease caused by Mycobacterium tuberculosis complex (MTBC) members, affecting several domestic animals, wildlife species and humans. The preliminary investigation was aimed to detect antibody against MTBC among indigenous wildlife which are free-ranged wild boar, free-ranged wild macaques and captive Asian elephants in selected areas of Selangor and elephant conservation centre in Pahang, respectively. The results indicate that MTBC serodetection rate in wild boar was 16.7% (7.3–33.5 at 95% confidence interval (CI)) using an in-house ELISA bPPD IgG and 10% (3.5–25.6 at 95% CI) by DPP^®VetTB assay, while the wild macaques and Asian elephant were seronegative. The univariate analysis indicates no statistically significant difference in risk factors for sex and age of wild boar but there was a significant positive correlation (P<0.05) between bovine TB in dairy cattle and wild boar seropositivity in the Sepang district.     

Phylogenetic analysis of Newcastle disease viruses isolated from wild birds in the Poyang Lake region of China

Newcastle disease virus (NDV) causes a highly contagious viral disease in poultry and wild birds, and it can cause significant economic loss worldwide. Eight viral strains were isolated by inoculating embryonated chicken eggs from the Poyang Lake region of China with swab samples. All eight of the NDV isolates were identified as class I genotype 3 strains, but they diverged notablely from class II viruses. Further analysis revealed that all eight NDV isolates were lentogenic strains containing the ¹¹²ERQER↓L¹¹⁷ motif at the F protein cleavage site. The strains were highly identical and were more species specific (chicken and waterfowl) than site specific (Nanchang and Duchang regions). The close phylogenetic proximity of these isolates indicates that viral transmission may happen between poultry and wild birds. Our study demonstrates that lentogenic class I NDVs exist in clinically healthy wild waterfowl and poultry within the Poyang Lake region. Active surveillance of these viruses to determine their evolution and origin is one of the most realistic strategies for preventing and controlling NDV outbreaks.     

北京郊区猪戊型肝炎流行初步调查

用间接ELISA试剂盒对北京市郊区所采集的174份猪血清样品进行了抗体检测,结果显示,其抗体总阳性率达62.1%。针对戊型肝炎病毒(HEV)ORF2/ORF3重叠区设计了简并引物,采用巢式RT-PCR对随即采取的少量猪粪便样品进行了抗原检测。结果表明,部分粪便样品中存在HEV。     

广东省两株猪源戊型肝炎病毒的分离鉴定及其全基因序列分析

为进一步了解广东省近年猪戊型肝炎病毒(HEV)基因组特点及其演化情况,本研究于2010年～2011年间从广东省不同地区猪场收集的69份病猪胆汁样品中,分离得到两株猪源HEV(命名为SS19和ZS11).采用通用引物通过PCR方法扩增病毒的全基因序列.应用DNAStar和MEGA 5基因分析软件,将分离株与24株不同基因型的HEV参考序列进行核苷酸全序列比对和遗传进化分析.结果显示,分离的两株猪源HEV SS19和ZS11的核苷酸序列相似性为95.6％,两个病毒分离株与基因Ⅳ型HEV各参考病毒株核苷酸序列最为接近,其相似性分别为83.2 ％～94.5％和83.6 ％～94.6％,并且均属于基因Ⅳ型HEV.实验结果表明,广东省HEV流行病毒株仍以基因Ⅳ型为主,病毒株之间存在一定的地域局限性.     

A systematic review and meta‐analysis of predictors of human hepatitis E virus exposure in non‐endemic countries

The reported incidence of clinical hepatitis E cases is rising in some non‐endemic countries, with concurrent concerns regarding potential hepatitis E virus (HEV) contamination of the blood supply. Therefore, the characterization of major potential sources of human HEV exposure is important to inform risk assessment and public health policy. A systematic review was conducted, including a comprehensive search in six electronic bibliographic databases, verified by hand‐searching reference lists of HEV reviews, and a grey literature search, of the broad research question ‘what is the evidence of the association between predictors of human HEV exposure, and HEV IgG seropositivity, in non‐endemic countries?’ Using forms designed a priori, captured studies were appraised at first‐level screening, second‐level characterization, and third‐level data extraction and risk of bias assessment. Meta‐analysis yielded summary estimates of association between potential predictors and odds of HEV seropositivity. Meta‐analysis and meta‐regression of the odds of HEV seroprevalence in specific groups characterized potential sources of HEV exposure. From 4,163 captured citations, 245 relevant studies underwent data extraction, investigating HEV seroprevalence or predictors in both healthy subjects and targeted patient groups. Across these groups, increasing age was a predictor of HEV IgG seropositivity. Both human immunodeficiency virus patients and haemodialysis patients had significantly increased odds of HEV seropositivity relative to the general population. Working with pigs, in forestry, or in hospitals, was significantly associated with increased odds of HEV seropositivity, as were consumption of meat, pork or game meat, or hunting. Chronological time was not associated with HEV seropositivity within our data sets. Further study of the distribution of potential dietary or behavioural predictors between high and lower prevalence areas within non‐endemic countries could improve our understanding of the relative importance of specific HEV transmission pathways.     

屠宰猪肝和血清中乙型肝炎病毒及戊型肝炎病毒的检测

应用1对乙型肝炎病毒（HBV）S基因保守区的引物，采用PCR方法从屠宰猪肝、血清中检测到了HBV，序列分析表明，扩增片段与已发表的HBVS基因的同源性高达98％～100％。电镜负染色样品观察结果表明，在HBV表面抗原ELISA检测强阳性反应的血清样品中存在有形态、大小与人HBV Dane颗粒和小球状颗粒相似的病毒粒子。针对戊型肝炎病毒（HEV）（）RF2／ORF3重叠区设计了简并引物，采用巢式RT-PCR对屠宰猪肝和血清样品进行了检测。结果表明，部分屠宰猪肝中存在HEV。