Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses

Dear Editor,The 2002–2003 global pandemic caused by severe acute respiratory syndrome coronavirus(SARS-CoV)infected around 8,000 people with 10%mortality(http://www.who.int/csr/sars/en/).The virus has a positivestranded RNA genome that encodes a large polyprotein(1a and 1ab),four structural proteins,including spike(S),

A bat MERS-like coronavirus circulates in pangolins and utilizes human DPP4 and host proteases for cell entry

It is unknown whether pangolins,the most trafficked mammals,play a role in the zoonotic transmission of bat coronaviruses.We report the circulation of a novel MERS-like coronavirus in Malayan pangolins,named Manis javanica HKU4-related coronavirus(MjHKU4r-CoV).Among 86 animals,four tested positive by pan-CoV PCR,and seven tested seropositive(11 and 12.8%).

Bat SARS coronavirus HKU3-1病毒株基因组及蛋白质的生物信息学分析

目的比较中国菊头蝠SARS冠状病毒(Bat SARS coronavirus)与人类及果子狸体内分离出来的SARS-Cov,分析它们是否存在一定的相似性。方法本文分析了Bat SARS coronavirus与已知冠状病毒Human SARS-CoV和civet SARS-CoV的系统进化关系,绘制了Bat SARS coronavirusHKU3-1的全基因序列图,同时预测了S蛋白和N蛋白的三维模型。结果对Bat SARS coronavirus HKU3-1的序列分析表明中国菊头蝠SARS-CoV与人和果子狸SARS-CoV非常相近。系统进化分析法显示Bat-SARS-CoV与SARS-CoV形成一个不同的群,称为group 2b CoV,与已知的group 2 CoV存在一定的距离。结论Bat SARS coronavirus HKU3-1不太可能是从人SARS-CoV传播过来的。另外,Bat SARS coronavirus与civet SARS-CoV可能具有共同的祖先,使得中国菊头蝠SARS病毒株同样具有感染人体的危险。

Severe acute respiratory syndrome (SARS) related coronavirus in bats

Three major human coronavirus disease outbreaks,severe acute respiratory syndrome(SARS),Middle East respiratory syndrome(MERS)and 2019 coronavirus disease(COVID-19),occurred in the twenty-first century and were caused by different coronaviruses(CoVs).All these viruses are considered to have originated from bats and transmitted to humans through intermediate hosts.SARS-CoV-1 and SARS-CoV-2,disease agent of COVID-19,shared around 80%genomic similarity,and thus belong to SARS-related CoVs.As a natural reservoir of viruses,bats harbor numerous other SARS-related CoVs that could potentially infect humans around the world,causing SARS or COVID19 like outbreaks in the future.In this review,we summarized the current knowledge of CoVs on geographical distribution,genetic diversity,cross-species transmission potential and possible pathogenesis in humans,aiming for a better understanding of bat SARS-related CoVs in the context of prevention and control.

A novel bat coronavirus with a polybasic furin-like cleavage site

The current pandemic of COVID-19 caused by a novel coronavirus,severe acute respiratory syndrome coronavirus-2(SARS-CoV-2),threatens human health around the world.Of particular concern is that bats are recognized as one of the most potential natural hosts of SARS-CoV-2;however,coronavirus ecology in bats is still nascent.Here,we performed a degenerate primer screening and next-generation sequencing analysis of 112 bats,collected from Hainan Province,China.Three coronaviruses,namely bat betacoronavirus(Bat CoV)CD35,Bat CoV CD36 and bat alpha-coronavirus CD30 were identified.Bat CoV CD35 genome had 99.5%identity with Bat CoV CD36,both sharing the highest nucleotide identity with Bat Hp-betacoronavirus Zhejiang2013(71.4%),followed by SARS-CoV-2(54.0%).Phylogenetic analysis indicated that Bat CoV CD35 formed a distinct clade,and together with Bat Hp-betacoronavirus Zhejiang2013,was basal to the lineage of SARS-CoV-1 and SARS-CoV-2.Notably,Bat CoV CD35 harbored a canonical furin-like S1/S2 cleavage site that resembles the corresponding sites of SARS-CoV-2.The furin cleavage sites between CD35 and CD36 are identical.In addition,the receptor-binding domain of Bat CoV CD35 showed a highly similar structure to that of SARS-CoV-1 and SARS-CoV-2,especially in one binding loop.In conclusion,this study deepens our understanding of the diversity of coronaviruses and provides clues about the natural origin of the furin cleavage site of SARS-CoV-2.

Global patterns of phylogenetic diversity and transmission of bat coronavirus

Bats are reservoirs for multiple coronaviruses(Co Vs).However,the phylogenetic diversity and transmission of global bat-borne Co Vs remain poorly understood.Here,we performed a Bayesian phylogeographic analysis based on 3,594 bat Co V Rd Rp gene sequences to study the phylogenetic diversity and transmission of bat-borne Co Vs and the underlying driving factors.We found that host-switching events occurred more frequently forα-Co Vs than forβ-Co Vs,and the latter was highly constrained by bat phylogeny.Bat species in the families Molossidae,Rhinolophidae,Miniopteridae,and Vespertilionidae had larger contributions to the cross-species transmission of bat Co Vs.Regions of eastern and southern Africa,southern South America,Western Europe,and Southeast Asia were more frequently involved in cross-region transmission events of bat Co Vs than other regions.Phylogenetic and geographic distances were the most important factors limiting Co V transmission.Bat taxa and global geographic hotspots associated with bat Co V phylogenetic diversity were identified,and bat species richness,mean annual temperature,global agricultural cropland,and human population density were strongly correlated with the phylogenetic diversity of bat Co Vs.These findings provide insight into bat Co Vevolution and ecological transmission among bat taxa.The identified hotspots of bat Co V evolution and transmission will guide early warnings of bat-borne Co V zoonotic diseases.

Pangolin HKU4-related coronaviruses found in greater bamboo bats from southern China

Coronavirus(CoV)spillover originating from game animals,particularly pangolins,is currently a significant concern.Meanwhile,vigilance is urgently needed for coronaviruses carried by bats,which are known as natural reservoirs of many coronaviruses.In this study,we collected 729 anal swabs of 20 different bat species from nine locations in Yunnan and Guangdong provinces,southern China,in 2016 and 2017,and described the molecular characteristics and genetic diversity of alphacoronaviruses(αCoVs)and betacoronaviruses(βCoVs)found in these bats.Using RT-PCR,we identified 58(8.0%)bat CoVs in nine bat species from six locations.Furthermore,using the Illumina platform,we obtained two representative full-length genomes of the bat CoVs,namely TyRo-CoV-162275 and TyRo-CoV-162269.Sequence analysis showed that TyRo-CoV-162275 shared the highest identity with Malayan pangolin(Manis javanica)HKU4-related coronaviruses(MjHKU4r-CoVs)from Guangxi Province,whereas TyRo-CoV-162269 was closely related to HKU33-CoV discovered in a greater bamboo bat(Tylonycteris robustula)from Guizhou Province.Notably,TyRo-CoV-162275 has a putative furin protease cleavage site in its S protein and is likely to utilize human dipeptidyl peptidase-4(hDPP4)as a cell-entry receptor,similar to MERSCoV.To the best of our knowledge,this is the first report of a bat HKU4r-CoV strain containing a furin protease cleavage site.These findings expand our understanding of coronavirus geographic and host distributions.

Coexistence of multiple coronaviruses in several bat colonies in an abandoned mineshaft

Since the 2002–2003 severe acute respiratory syndrome(SARS) outbreak prompted a search for the natural reservoir of the SARS coronavirus, numerous alpha- and betacoronaviruses have been discovered in bats around the world. Bats are likely the natural reservoir of alpha- and betacoronaviruses, and due to the rich diversity and global distribution of bats, the number of bat coronaviruses will likely increase. We conducted a surveillance of coronaviruses in bats in an abandoned mineshaft in Mojiang County, Yunnan Province, China, from 2012–2013. Six bat species were frequently detected in the cave: Rhinolophus sinicus, Rhinolophus affinis, Hipposideros pomona, Miniopterus schreibersii, Miniopterus fuliginosus, and Miniopterus fuscus. By sequencing PCR products of the coronavirus RNA-dependent RNA polymerase gene(Rd Rp), we found a high frequency of infection by a diverse group of coronaviruses in different bat species in the mineshaft. Sequenced partial Rd Rp fragments had 80%–99% nucleic acid sequence identity with well-characterized Alphacoronavirus species, including Bt CoV HKU2, Bt CoV HKU8, and Bt CoV1,and unassigned species Bt CoV HKU7 and Bt CoV HKU10. Additionally, the surveillance identified two unclassified betacoronaviruses, one new strain of SARS-like coronavirus, and one potentially new betacoronavirus species. Furthermore, coronavirus co-infection was detected in all six bat species, a phenomenon that fosters recombination and promotes the emergence of novel virus strains. Our findings highlight the importance of bats as natural reservoirs of coronaviruses and the potentially zoonotic source of viral pathogens.

The persistent prevalence and evolution of cross-family recombinant coronavirus GCCDC1 among a bat population:a two-year follow-up

Bats are connected with the increasing numbers of emerging and re-emerging viruses that may break the species barrier and spread into the human population. Coronaviruses are one of the most common viruses discovered in bats, which were considered as the natural source of recent human-susceptible coronaviruses, i.e. SARS-COV and MERS-CoV. Our previous study reported the discovery of a bat-derived putative cross-family recombinant coronavirus with a reovirus gene p10, named as Ro-BatCoV GCCDC1. In this report, through a two-year follow-up of a special bat population in one specific cave of south China, we illustrate that Ro-BatCoV GCCDC1 persistently circulates among bats. Notably, through the longitudinal observation, we identified the dynamic evolution of Ro-BatCoV GCCDC1 in bats represented by continuously recombination events. Our study provides the first glimpse of the virus evolution in one longitudinally observed bat population cohort and underlines the surveillance and pre-warning of potential interspecies transmittable viruses in bats.

Human-animal interactions and bat coronavirus spillover potential among rural residents in Southern China

Human interaction with animals has been implicated as a primary risk factor for several high impact zoonoses,including many bat-origin viral diseases.However the animal-to-human spillover events that lead to emerging diseases are rarely observed or clinically examined,and the link between specific interactions and spillover risk is poorly understood.To investigate this phenomenon,we conducted biological-behavioral surveillance among rural residents in Yunnan,Guangxi,and Guangdong districts of Southern China,where we have identified a number of SARS-related coronaviruses in bats.Serum samples were tested for four bat-borne coronaviruses using newly developed enzyme-linked immunosorbent assays(ELISA).Survey data were used to characterize associations between human-animal contact and bat coronavirus spillover risk.A total of 1,596 residents were enrolled in the study from 2015 to 2017.Nine participants(0.6%)tested positive for bat coronaviruses.265(17%)participants reported severe acute respiratory infections(SARI)and/or influenza-like illness(ILI)symptoms in the past year,which were associated with poultry,carnivore,rodent/shrew,or bat contact,with variability by family income and district of residence.This study provides serological evidence of bat coronavirus spillover in rural communities in Southern China.The low seroprevalence observed in this study suggests that bat coronavirus spillover is a rare event.Nonetheless,this study highlights associations between human-animal interaction and zoonotic spillover risk.These findings can be used to support targeted biological behavioral surveillance in high-risk geographic areas in order to reduce the risk of zoonotic disease emergence.

Genetic diversity of coronaviruses in Miniopterus fuliginosus bats

Coronaviruses, such as severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome coronavirus, pose significant public health threats. Bats have been suggested to act as natural reservoirs for both these viruses, and periodic monitoring of coronaviruses in bats may thus provide important clues about emergent infectious viruses. The Eastern bent-wing bat Miniopterus fuliginosus is distributed extensively throughout China. We therefore analyzed the genetic diversity of coronaviruses in samples of M. fuliginosus collected from nine Chinese provinces during 2011–2013. The only coronavirus genus found was Alphacoronavirus. We established six complete and five partial genomic sequences of alphacoronaviruses, which revealed that they could be divided into two distinct lineages, with close relationships to coronaviruses in Miniopterus magnater and Miniopterus pusillus. Recombination was confirmed by detecting putative breakpoints of Lineage 1 coronaviruses in M. fuliginosus and M. pusillus(Wu et al., 2015), which supported the results of topological and phylogenetic analyses. The established alphacoronavirus genome sequences showed high similarity to other alphacoronaviruses found in other Miniopterus species, suggesting that their transmission in different Miniopterus species may provide opportunities for recombination with different alphacoronaviruses. The genetic information for these novel alphacoronaviruses will improve our understanding of the evolution and genetic diversity of coronaviruses, with potentially important implications for the transmission of human diseases.

Erratum to: Bat origin of a new human coronavirus: there and back again

We miscalculated identities of several genes, and the corrected Table 1 should be as follows:a、 This table demonstrates identities of 2019-nCoV WIV04 (GenBank Accession No. MN996528.1) compared with bat SARSr-CoV RaTG13 (GenBank Accession No. MN996532.1) and SARS-CoV BJ01 (GenBank Accession No. AY278488.2).

Interspecies transmission and evolution of the emerging coronaviruses: perspectives from bat physiology and protein spatial structure

Emergent coronaviruses(CoVs)such as SARS-CoV and MERS-CoV have posed great threats to public health worldwide over the past two decades.Currently,the emergence of SARS-CoV-2 as a pandemic causes greater public health concern.CoV diversity is due to the large size and replication mechanisms of the genomes together with having bats as their optimum natural hosts.The ecological behavior and unique immune characteristics of bats are optimal for the homologous recombination of CoVs.The relationship of spatial structural characteristics of the spike protein,a protein that is critical for recognition by host receptors,in different CoVs may provide evidence in explaining the coevolution of CoVs and their hosts.This information may help to enhance our understanding of CoV evolution and thus provide part of the basis of preparations for any future outbreaks.

SPA-ELISA检测棕果蝠血清冠状病毒抗体的研究

目的建立一种敏感、简便的检测蝙蝠血清冠状病毒抗体的方法。方法利用葡萄球菌A蛋白(SPA)能够与某些哺乳动物IgG抗体Fc段非特异性结合的特点,对市售人冠状病毒抗体ELISA试剂盒进行改造,建立SPA-ELISA法,使其适用于蝙蝠血清冠状病毒抗体的检测,并对采集到的55份棕果蝠血清冠状病毒抗体进行检测。结果用SPA-ELISA方法对人冠状病毒试剂盒中阳性与阴性对照、SARS病人恢复期血清、空白对照的检测都得到理想的结果,并从55份棕果蝠血清中检测出2例阳性,阳性率为3.64%(2/55),中和试验进一步证实了结果的真实性。结论所建立的方式适用于检测棕果蝠血清冠状病毒抗体。

广西岭景镇某山村蝙蝠携带SARS样冠状病毒及狂犬病毒的调查研究

目的了解广西岭景镇某山村蝙蝠携带SARS样冠状病毒及狂犬病毒的情况。方法对广西岭景镇某山村开展流行病学调查,捕捉蝙蝠,采集咽拭、肛拭及粪便标本,进行RT-PCR和细胞分离培养检测狂犬病毒(Rabies virus,RV)、SARS样冠状病毒(SARS-like Coronavirus,SARS-like-CoV)。结果本次调查收集了4只犬蝠(Cynopterus sphinx)、6只普氏蹄蝠(Hipposideros pratti,)和7只中菊头蝠(Rhinolophus affinis),在采集的标本中未检测到SARS样冠状病毒及狂犬病毒。结论未发现调查的3种蝙蝠携带SARS样冠状病毒及狂犬病毒。

广州及周边地区蝙蝠携带SRAS冠状病毒的流行病学调查

目的了解野生动物市场果蝠携带SARS冠状病毒(SARS-CoV)和SARS样冠状病毒(SARS-like-CoV)情况。方法2004年9月至2005年11月,在广州及广州野生动物市场收集来自广州及周边地区的分属两个亚目9个种的蝙蝠共905只,有犬蝠(Cynopterussphinx)、棕果蝠(Rousettusleschenaulti)、普通长翼蝠(Miniopterusschreibersi)、普氏蹄蝠(Hipposiderospratti)、中华菊头蝠(Rhinolophussinicus)、小黄蝠(Scotophiluskuhlii)、大耳双色蹄蝠(HipposiderosPomona)、中菊头蝠(Rhinolophusaffinis)和小菊头蝠(Rhinolophuspusillus)。收集的蝙蝠分别取得咽拭813份、血清524份、肺组织853份及直肠粪便853份标本共计3043份,用新型冠状病毒(N-蛋白)测定试剂盒(酶联免疫法)、SARS-CoVRNA检测试剂盒、荧光PCR及基因芯片检测并通过RT-PCR和细胞分离培养检测SARS-CoV和SARS-like-CoV。结果收集的9个种905只蝙蝠共3043份标本未检测、分离出SARS-CoV和SARS-like-CoV。结论调查的广州及周边地区的9种蝙蝠均未检测到SARS-CoV和SARS-like-CoV。

蝙蝠SARS样冠状病毒S蛋白受体结合域在昆虫细胞中的表达和纯化

目的:在昆虫细胞表达系统中表达蝙蝠SARS样冠状病毒Spike蛋白的受体结合域,并探讨其表达动力学和纯化条件。方法:以PCR扩增蝙蝠SARS样冠状病毒Spike基因的受体结合域片段,产物连接到pMD-T载体,再亚克隆至供体质粒pFAST-HTB,经序列测定确认基因正确克隆,进一步将其转座入Bacmid中,在昆虫细胞Sf9中进行表达,采用SDS-PAGE和Western blotting对表达产物进行分析,并通过镍离子螯合树脂纯化重组蛋白。结果:在昆虫细胞表达系统中表达出蝙蝠SARS样冠状病毒Spike蛋白的受体结合域片段,当感染复数(multiplicity of infectiin,MOI)为2时,重组病毒感染细胞56 h后,重组蛋白的表达量达到峰值。结论:蝙蝠SARS样冠状病毒Spike蛋白的受体结合域片段在昆虫细胞表达系统中得到了有效表达,并可通过镍离子螯合树脂纯化重组蛋白。

新型冠状病毒(SARS-CoV-2)传播的溯源分析及启示

由新型冠状病毒SARS-CoV-2(severe acute respiratory syndrome coronavirus 2)感染引起的疫情严重危害人类的生命安全,给社会造成巨大经济损失。目前,SARS-CoV-2从源头到感染人类过程中,哪些动物充当了传播媒介的角色尚未有定论。找到SARSCoV-2的动物宿主来源并切断传播途径是防控传染病的有效措施,对预防疫情再次发生、保障社会公共卫生安全具有重要意义。该文总结了SARS-CoV-2传播宿主的最新研究进展,并依据疫情事实推测SARS-CoV-2从蝙蝠到人类可能存在多个中间宿主。因此,在当前疫情防控乃至未来对新发传染病的防控中,基于同一健康创新理念加强人类医学-动物医学一体化防控体系建设至关重要。

蝙蝠在冠状病毒传播中的作用

严重性急性呼吸系统综合症(severe acute respiratory syndrome,SARS),又名非典型性肺炎,虽爆发已经10年过去了,恐惧依旧笼罩在人们心中,近日,中东呼吸系统综合症(middle east respiratory syndrome,MERS)又强势来袭,再次引发了人们对冠状病毒的强烈关注。近年来研究证明,蝙蝠在冠状病毒的传播中具有重要作用。本文对冠状病毒的多样性和蝙蝠在冠状病毒传播中作用进行了比较系统综述。

蝙蝠相关的冠状病毒

2019年12月,新型冠状病毒肺炎(COVID-19)暴发,继2003年严重急性呼吸综合征(SARS)暴发后又给人类敲响了一记警钟。研究者通过溯源分析,发现COVID-19病原体严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的自然宿主可能是中华菊头蝠。21世纪以来,全球共出现4次冠状病毒[严重急性呼吸综合征冠状病毒(SARS-CoV)、中东呼吸综合征冠状病毒(MERS-CoV)、猪急性腹泻综合征冠状病毒(SADS-CoV)、SARS-CoV-2]的暴发,给人类健康、公共卫生、经济发展及社会稳定造成了巨大的威胁与损失。大量证据表明,这4种冠状病毒的自然宿主可能均为蝙蝠。本文对蝙蝠相关冠状病毒的种类、全球地理分布及引起暴发的蝙蝠相关冠状病毒作一综述。