Abstract
We collected 5,500 mosquitoes belonging to six species in three locations in China. Their viromes were tested using metagenomic sequencing and bioinformatic analysis. The affluent viral sequences that were detected and annotated belong to 22 viral taxonomic families. Then, PCR was performed to confirm the results, followed by phylogenetic analysis. Herein, part of mosquito virome was identified, including chikungunya virus (CHIKV), Getah virus (GETV), and Ross river virus (RRV). After metagenomic analysis, seven CHIKV sequences were verified by PCR amplification, among which CHIKV-China/YN2018-1 had the highest homology with the CHIKV isolated in Senegal, 1983, with a nucleotide (nt) identity of at least 81%, belonging to genotype West Africa viral genes. Five GETV sequences were identified, which had a high homology with the GETV sequences isolated from Equus caballus in Japan, 1978, with a (nt) identity of at least 97%. The newly isolated virus CHIKV-China/YN2018-1 became more infectious after passage of the BHK-21 cell line to the Vero cell line. The newly identified RRV gene had the highest homology with the 2006 RRV isolate from Australia, with a (nt) identity of at least 94%. In addition, numerous known and unknown viruses have also been detected in mosquitoes from Yunnan province, China, and propagation tests will be carried out.
Highlights
Mosquitoes are important biological vectors of many infectious viruses, including Getah virus (GETV), Ross River virus (RRV), chikungunya virus (CHIKV), Zika virus (ZIKV), and dengue virus (DENV), which were a menace to global public health and security (Sinclair and Asgari, 2020; Nowee et al, 2021; Pyke et al, 2021)
Mosquitoes are the intermediate host of many viruses, playing an important part in the spread of numerous viral infectious diseases (Zhu et al, 2019; Estrada-Franco et al, 2020)
Compared with the traditional method, the efficiency of metagenomic sequencing by Illumina sequencing combined with bioinformatics analysis is higher, and in this way, many viruses from different species have been identified in recent years (Zhang et al, 2018; Marzoli et al, 2021; Thannesberger et al, 2021)
Summary
Mosquitoes are important biological vectors of many infectious viruses, including Getah virus (GETV), Ross River virus (RRV), chikungunya virus (CHIKV), Zika virus (ZIKV), and dengue virus (DENV), which were a menace to global public health and security (Sinclair and Asgari, 2020; Nowee et al, 2021; Pyke et al, 2021). Mosquitoes were considered as an intermediate host of numerous viruses that infect humans, animals, insects, plants, and other species (de Boer et al, 2021). Once mosquitoes bite the host with viremia, they will be infected, and viruses can replicate and reproduce in the mosquitoes, followed by spread to more victims during the bite and blood-sucking process (Garcia et al, 2018; Martıń ez et al, 2020). Reverse transcription polymerase chain reaction (RT-PCR) and nested PCR methods were generally employed for detecting mosquito-borne viruses (Cabral-Castro et al, 2016; Morales et al, 2017), while these conventional methods are time-consuming and labor-intensive in the detection of low-level mosquito viromes, compared to Illumina sequencing (He et al, 2013; Martıń ez-Puchol et al, 2020). Metagenomic analysis may have important value in avoiding missing detection of highly infectious and pathogenic viruses in different mosquito species, as well as unknown viruses
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