Transmission competence of a new mesonivirus, Yichang virus, in mosquitoes and its interference with representative flaviviruses.

Advances in technology have greatly stimulated the understanding of insect-specific viruses (ISVs). Unfortunately, most of these findings are based on sequencing technology, and laboratory data are scarce on the transmission dynamics of ISVs in nature and the potential effects of these viruses on arboviruses. Mesonivirus is a class of ISVs with a wide geographical distribution. Recently, our laboratory reported the isolation of a novel strain of mesonivirus, Yichang virus (YCV), from Culex mosquitoes, China. In this study, the experimental infection of YCV by the oral route for adult and larvae mosquitoes, and the vertical transmission has been conducted, which suggests that YCV could adopt a mixed-mode transmission. Controlled experiments showed that the infectivity of YCV depends on the mosquito species, virus dose, and infection route. The proliferation curve and tissue distribution of YCV in Cx. quinquefasciatus and Ae. albopictus showed that YCV is more susceptible to Ae. albopictus and is located in the midgut. Furthermore, we also assessed the interference of YCV with flaviviruses both in vitro and in vivo. YCV significantly inhibited the proliferation of DENV-2 and ZIKV, in cell culture, and reduced transmission rate of DENV-2 in Ae. albopictus. Our work provides insights into the transmission of ISVs in different mosquito species during ontogeny and their potential ability to interact with mosquito-borne viruses.

Vector competence of anthropophilic mosquitoes for a new mesonivirus in Senegal.

The mesoniviruses (MESOVs) belong to the newly described Mesoniviridae family (Order: Nidovirales). They have never been reported in Senegal until recently during a study in arbovirus emergence with the detection of a new species of MESOV named Dianke virus (DKV) from common mosquitoes from eastern Senegal. Actually, their vector competence for this newly described DKV is unknown. We, therefore, estimated the vector competence of Culex tritaeniorhynchus, Culex quinquefasciatus, Aedes aegypti, and Anopheles gambiae mosquitoes collected in Senegal for DKV using oral infection. Whole bodies, legs/wings, and saliva samples were tested for DKV by RT-PCR to estimate infection, dissemination, and transmission rates. The infectivity of virus particles in the saliva was confirmed by infecting C6/36 cells. Virus transmission rates were up to 95.45% in Culex tritaeniorhynchus, 28% in Cx. quinquefasciatus and 9.09% in Aedes aegypti. Viral particles in the saliva were confirmed infectious by C6/36 cell culture. An. gambiae was able to disseminate DKV only at 20 days post-infection. This study shows that Culex mosquitoes are more competent than Ae. aegypti for DKV, while Anopheles gambiae is not likely a competent vector.

[Isolation and Identification of the Nam Dinh Virus from Mosquitoes on the China-Laos-Myanmar Border].

Three strains of an insect nidovirus, the Nam Dinh virus (NDiV), isolated in Yunnan Province, China, have been identified. Aedes albopictus C6/36 cells were used to isolate NDiV from mosquitoes collected in Yunnan Province in 2012.Culture supernatants with a positive cytopathic effect were harvested for virus identification by sequence-independent single primer amplification. Transmission electron microscopy revealed virion structure to be spherical with a diameter of 60~80nm.Reverse transcription-polymerase chain reaction was applied to amplify sequences of RNA-dependent RNA-polymerase (RdRp), HEL1(superfamily 1helicase)and spike protein. The amino-acid sequences of three isolates from Yunnan Province showed>98% homology with NDiV strains. Phylogenetic analyses showed that these three isolates, along with NDiV, could be classified into the family Mesoniviridae.