Abstract
BackgroundThe re-emergence of chikungunya (CHIK) fever in Thailand has been caused by a novel lineage of chikungunya virus (CHIKV) termed the Indian Ocean Lineage (IOL). The Aedes albopictus mosquito is thought to be a primary vector of CHIK fever in Thailand, whereas Ae. aegypti acts as a secondary vector of the virus. The vertical transmission is believed to be a primary means to maintain CHIKV in nature and may be associated with an increased risk of outbreak. Therefore, the goal of this study was to analyze the potential of these two Thai mosquito species to transmit the virus vertically and to determine the number of successive mosquito generations for the virus transmission.MethodsTwo-hundred-and-fifty female Ae. aegypti and Ae. albopictus mosquitoes were artificially fed a mixture of human blood and CHIKV IOL. Mosquito larvae and adults were sampled and screened for CHIKV by one-step qRT-PCR. LLC-MK2 cell line was used to isolate CHIKV in the mosquitoes each generation. The virus isolate was identified by immunocytochemical staining and was confirmed by sequencing. Both mosquito species fed on human blood without CHIKV and uninfected LLC-MK2 cells were used as controls.ResultsAedes aegypti and Ae. albopictus mosquitoes were able to transmit CHIKV vertically to F5 and F6 progenies, respectively. The virus isolated from the two mosquito species caused cytopathic effect in LLC-MK2 cells by 2 days post-infection and immunocytochemical staining showed the reaction between CHIKV IOL antigen and specific monoclonal antibody in the infected cells. DNA sequence confirmed the virus transmitted vertically as CHIKV IOL with E1-A226V mutation. No CHIKV infection was observed in both mosquito species and LLC-MK2 cells from control groups.ConclusionsThe study demonstrated that Ae. aegypti and Ae. albopictus mosquitoes from Thailand are capable of transmitting CHIKV IOL vertically in the laboratory. Our results showed that Ae. albopictus is more susceptible and has a greater ability to transmit the virus vertically than Ae. aegypti. This knowledge would be useful for risk assessments of the maintenance of CHIKV in nature, which is crucial for disease surveillance, vector control and the prevention of potential CHIKV epidemics.
Highlights
The re-emergence of chikungunya (CHIK) fever in Thailand has been caused by a novel lineage of chikungunya virus (CHIKV) termed the Indian Ocean Lineage (IOL)
Vertical transmission rates in Ae. aegypti and Ae. albopictus CHIKV IOL with the A226V mutation in E1 gene used in this experiment was confirmed by sequencing (Fig. 1)
The experiment revealed that CHIKV IOL was detectable until the F5 generation of Ae. aegypti larvae and adults, whereas the persistence of the virus in Ae. albopictus larvae and adults was found to be to the F6 generation
Summary
The re-emergence of chikungunya (CHIK) fever in Thailand has been caused by a novel lineage of chikungunya virus (CHIKV) termed the Indian Ocean Lineage (IOL). The vertical transmission is believed to be a primary means to maintain CHIKV in nature and may be associated with an increased risk of outbreak. In Africa, CHIKV is primarily maintained in a sylvatic, enzootic cycle, which involves non-human primates as reservoirs/ amplifying hosts and arboreal, primatophilic Aedes mosquito species as vectors (Ae. furcifer-taylori, Ae. africanus, Ae. luteocephalus, Ae. neoafricanus) [2]. CHIKV maintenance in Asia is largely through an endemic/epidemic cycle, in which humans serve as the primary hosts with Ae. aegypti historically serving as the primary vector [3]. CHIKV has been previously documented to cause outbreaks with three distinct genotypes based on the E1 envelope glycoprotein sequences: the West African genotype, the East, Central and South African (ECSA) genotypes, and the Asian genotype [4]
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