27The ongoing outbreak of viral pneumonia in China and beyond is associated with a novel 28 coronavirus, provisionally termed 2019-nCoV. This outbreak has been tentatively associated 29 with a seafood market in Wuhan, China, where the sale of wild animals may be the source of 30 zoonotic infection. Although bats are likely reservoir hosts for 2019-nCoV, the identity of 31 any intermediate host facilitating transfer to humans is unknown. Here, we report the 32 identification of 2019-nCoV related coronaviruses in pangolins (Manis javanica) seized in 33 anti-smuggling operations in southern China. Metagenomic sequencing identified pangolin 34 associated CoVs that belong to two sub-lineages of 2019-nCoV related coronaviruses, 35 including one very closely related to 2019-nCoV in the receptor-binding domain. The 36 discovery of multiple lineages of pangolin coronavirus and their similarity to 2019-nCoV 37 suggests that pangolins should be considered as possible intermediate hosts for this novel 38 human virus and should be removed from wet markets to prevent zoonotic transmission.
Highlights d Six high-quality ixodid tick genomes and 678 re-sequenced tick specimens d Insights into the genetic basis of tick hematophagy and related phenotypes d Population structure and genetic diversity of six tick species d Tick-borne pathogen composition and distribution by metagenome analyses
Background A tick-borne segmented RNA virus called Jingmen tick virus (JMTV) was recently identified, variants of which were detected in a non-human primate host and fatal patients with Crimean-Congo haemorrhagic fever. We investigated its infectivity and pathogenicity for humans. Methods We obtained skin-biopsy, blood and serum samples from patients with tick bites, and used high-throughput sequencing, in situ hybridisation, and serologic testing to diagnose and ascertain the cases of JMTV infection. Findings A JMTV strain was isolated from the tick Amblyomma javanense into an embryo-derived tick cell line. We obtained sustained passage of JMTV, and revealed that it was able to accumulate in salivary glands of experimentally infected ticks. Four JMTV-infected patients were identified by high-throughput sequencing of skin biopsies and blood samples. The virus replication in skin tissue was visualised by in situ hybridisation. The four patients all had an itchy or painful eschar at the site of tick bite, with or without lymphadenopathy. Immunohistochemical examination revealed remarkable local inflammation manifested as infiltration by neutrophils. Eight patients were identified by serological testing and showed more severe clinical manifestations. Two Ixodes persulcatus ticks detached from patients were positive for JMTV. All JMTV strains identified in this study formed a well-supported sub-lineage, distinct from those previously reported in China. Interpretation The public significance of JMTV should be highly concerning due to its potential pathogenicity for humans and efficient transmission by potential ticks. Fund China Natural Science Foundation, State Key Research Development Programme, and United Kingdom Biotechnology and Biological Sciences Research Council.
We used molecular methods to identify Rickettsia raoultii infections in 2 persons in China. These persons had localized rashes around sites of tick bites. R. raoultii DNA was detected in 4% of Dermacentor silvarum ticks collected in the same area of China and in 1 feeding tick detached from 1 patient.
Spotted fever group (SFG) rickettsiae are important causative agents of (re)emerging tick‐borne infectious diseases in humans, and ticks play a key role in their maintenance and transmission. In this study, hard ticks were collected from five sampling sites in North China in 2017 and 2018. Of them, Haemaphysalis longicornis, Rhipicephalus microplus and Dermacentor nuttalli were collected from livestock (sheep and goats) and the vegetation, Hyalomma asiaticum from sheep, goats and camels, and Hyalomma marginatum from sheep and goats. The SFG rickettsiae were identified in these ticks by amplifying the partial rrs and complete 17‐kDa genes, with an overall infection rate of 52.9%. In addition, the nearly full‐length rrs and gltA and partial ompA genes were recovered to classify the species of SFG rickettsiae further. Phylogenetic analysis revealed the presence of three human pathogenic species in Hy. asiaticum, Hy. marginatum, Ha. longicornis and De. nuttalli, including two cultured ones (Rickettsia raoultii and Rickettsia aeschlimannii) and one uncultured (Candidatus R. jingxinensis). Furthermore, partial groEL gene was also obtained, and phylogenetic trees were also reconstructed to better understand the genetic relationship with known sequences in each SFG rickettsiae species detected in the current study. Notably, the R. aeschlimannii sequences described in this study were closely related to those from abroad rather than from another part of China, indicating their different origin. However, the R. raoultii and Ca. R. jingxinensis sequences presented close relationship with variants from other parts of China. In sum, our data revealed SFG rickettsiae species in northern China, highlighting the need for surveillance of their infection in local humans.
Background Four species within Anaplasma genus are emerging zoonotic pathogens, which are transmitted by ticks and generate veterinary and public health concerns. Here, we performed a molecular survey of Anaplasma in Ankang, Northwest China. Methods Hard ticks were collected and identified using morphological and molecular methods. Human-pathogenic Anaplasma species were tested using nested polymerase chain reaction. The nearly complete rrs , gltA , and groEL genes sequences from revealed Anaplasma species were amplified and sequenced to determine their molecular characteristics and their phylogeny. Results All ticks collected in Ankang belonged to the Rhipicephalus microplus . Novel unclassified Anaplasma strains genetically related to A. platys and A. capra were detected in these ticks. Co-infection of these two organisms was also found. The novel unclassified Anaplasma strains identified in this study formed a distinct phylogenetic lineage based on the groEL gene and two lineages based on the gltA gene within A. platys and related strains group. The revealed A. capra strains identified in this study were most closely related to those detected in humans and other vertebrate animals. Conclusion We revealed the presence of A. capra , a novel human pathogens in R. microplus ticks in previously unrecognized endemic regions. We also detected a novel unclassified Anaplasma species genetically related to A. platys . The epidemiology of anaplasmosis caused by these two Anaplasma species in humans should be assessed in future studies. Electronic supplementary material The online version of this article (10.1186/s12879-019-4075-3) contains supplementary material, which is available to authorized users.
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