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. 2022 Mar 3:13:791563.
doi: 10.3389/fmicb.2022.791563. eCollection 2022.

First Detection of Mukawa Virus in Ixodes persulcatus and Haemaphysalis concinna in China

Yu-Na Wang  1 , Rui-Ruo Jiang  2 , Heng Ding  1 , Xiao-Long Zhang  3 , Ning Wang  1 , Yun-Fa Zhang  1 , Yue Li  1 , Jin-Jin Chen  1 , Pan-He Zhang  1 , Hao Li  1 , Jia-Fu Jiang  1 , Lan-Zheng Liu  4 , Meng-Bin Yu  2 , Gang Wang  1 , Xiao-Ai Zhang  1 , Wei Liu  1
Affiliations

First Detection of Mukawa Virus in Ixodes persulcatus and Haemaphysalis concinna in China

Yu-Na Wang et al. Front Microbiol. .

Abstract

Mukawa virus (MKWV), a novel tick-borne virus (TBV) of the genus Phlebovirus of family Phenuiviridae, has been firstly reported in Ixodes persulcatus in Japan. In this study, we made an epidemiological investigation in China to obtain the geographic distribution and genetic features of this virus outside Japan. We screened 1,815 adult ticks (665 I. persulcatus, 336 Dermacentor silvarum, 599 Haemaphysalis longicornis, 170 Rhipicephalus microplus, 45 Haemaphysalis concinna) and 805 wild small mammals collected from eight provinces. The positive rate of 6.77% (45/665, including 18 female and 27 male I. persulcatus) and 2.22% (1/45, 1 male H. concinna) were obtained from I. persulcatus and H. concinna in Heilongjiang province, respectively. No evidence of MKWV infection was found in other three tick species or any of the mammalian species. The virus can infect the Vero cells successfully, indicating the ability of MKWV to replicate in mammalian cells. A phylogenetic tree based on the nucleotide sequences of L, M, and S segments demonstrated that the Japanese MKWV variant, our two MKWV variants, and KURV were clustered with the members of the mosquito/sandfly-borne phleboviruses and distant from other tick-borne phenuiviruses. A phylogenetic analysis based on 895 bp partial L gene sequences (n = 46) showed that all MKWV sequences were separated into three lineages. Our results showed the presence of MKWV in I. persulcatus and H. concinna in northeast of China, highlighting the necessity of epidemiological study in wider regions. Due to the ability of MKWV to replicate in mammalian cells, the potential for zoonosis, and wide distribution of I. persulcatus and H. concinna in China, the important vectors of MKWV, further screening to more tick species, wild animals, domestic animals, and humans raises up practical significance.

Keywords: China; Haemaphysalis concinna; Ixodes persulcatus; tick-borne phlebovirus; ticks.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Map of China showing the collection sites for ticks and wild small mammals for MKWV detection. The sampling number for each tick was shown in parentheses; the sampling number for each animal species was marked above the column.
FIGURE 2
FIGURE 2
Comparison of homology between MKWV and KURV. Nucleotide and amino acid identities between Japanese MKWV, KURV, and our MKWV from I. persulcatus are marked with red. Nucleotide and amino acid identities’ sequences between Japanese MKWV, KURV, and our MKWV from H. concinna are marked with purple. The nucleotide and amino acid identities of our two MKWVs are marked with blue.
FIGURE 3
FIGURE 3
Phylogenetic analysis of MKWV RNA segments. (A) The ML phylogenetic tree constructed based on a 5,672-bp fragment of the partial L segment. (B) The ML phylogenetic tree constructed based on the full-length nucleotide sequences of the M segment. (C) The ML phylogenetic tree constructed based on the full-length nucleotide sequences of the S segment. Our MKWV from I. persulcatus was labeled with a red solid five-pointed star, and our MKWV from H. concinna was labeled with a green solid five-pointed star. Trees that were generated using MEGA-X and analyzed included 1,000 bootstrap replicates.
FIGURE 4
FIGURE 4
Phylogenetic analysis of MKWV N and NSs proteins. (A) The maximum likelihood phylogenetic tree based on the deduced amino acid sequences of the MKWV nucleoprotein protein (N). (B) The maximum likelihood phylogenetic tree based on the deduced amino acid sequences of the MKWV non-structural protein (NSs). Our MKWV from I. persulcatus was labeled with a red solid five-pointed star, and our MKWV from H. concinna was labeled with a green solid five-pointed star. Trees that were generated using MEGA-X and analyzed included 1,000 bootstrap replicates.
FIGURE 5
FIGURE 5
Phylogenetic analysis of 895-bp MKWV-positive RNA sequences. The Maximum likelihood phylogenetic tree constructed based on the 895-bp MKWV-positive RNA sequences (n = 46) of the L segment. The MKWV-positive RNA sequences from I. persulcatus were labeled with red solid dots, and the MKWV-positive RNA sequences from H. concinna were labeled with green solid dots. Trees that were generated using MEGA-X and analyzed included 1,000 bootstrap replicates.
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