Abstract
BackgroundEmerging and re-emerging arthropod-borne viruses (arboviruses) cause human and animal disease globally. Field and laboratory investigation of mosquito-borne arboviruses requires analysis of mosquito samples, either individually, in pools, or a body component, or secretion such as saliva. We assessed the applicability of mosquito excreta as a sample type that could be utilized during studies of Ross River and West Nile viruses, which could be applied to the study of other arboviruses.Methodology/Principal findingsMosquitoes were fed separate blood meals spiked with Ross River virus and West Nile virus. Excreta was collected daily by swabbing the bottom of containers containing batches and individual mosquitoes at different time points. The samples were analyzed by real-time RT-PCR or cell culture enzyme immunoassay. Viral RNA in excreta from batches of mosquitoes was detected continuously from day 2 to day 15 post feeding. Viral RNA was detected in excreta from at least one individual mosquito at all timepoints, with 64% and 27% of samples positive for RRV and WNV, respectively. Excretion of viral RNA was correlated with viral dissemination in the mosquito. The proportion of positive excreta samples was higher than the proportion of positive saliva samples, suggesting that excreta offers an attractive sample for analysis and could be used as an indicator of potential transmission. Importantly, only low levels of infectious virus were detected by cell culture, suggesting a relatively low risk to personnel handling mosquito excreta.Conclusions/SignificanceMosquito excreta is easily collected and provides a simple and efficient method for assessing viral dissemination, with applications ranging from vector competence experiments to complementing sugar-based arbovirus surveillance in the field, or potentially as a sample system for virus discovery.
Highlights
It has been estimated that vector-borne diseases account for almost 20% of the global burden of infectious diseases, with more than 80% of the world’s population living in areas at risk [1]
We assessed the applicability of mosquito excreta as a sample type that could be utilized during studies of Ross River and West Nile viruses, which could be applied to the study of other arboviruses
Testing for the presence of arboviruses in mosquitoes used in laboratory experiments or surveillance usually involves collecting samples, from pools of hundreds of mosquitoes to the legs and wings of an individual mosquito and testing them by different methods
Summary
It has been estimated that vector-borne diseases account for almost 20% of the global burden of infectious diseases, with more than 80% of the world’s population living in areas at risk [1]. Vector competence refers to the ability of a mosquito or other hematophagous arthropod to acquire, replicate, and successfully transmit a pathogen [5] This is a key parameter to estimate vectorial capacity, namely the potential of a mosquito population to transmit an infectious agent to a susceptible host population [6]. Transmission can be assessed in vitro, by forcing mosquitoes to salivate into capillary tubes [11] and testing the expectorate for virus by inoculation in cell culture or by molecular assays This method is relatively simple and removes ethical and logistical issues with working with live vertebrates. We assessed the applicability of mosquito excreta as a sample type that could be utilized during studies of Ross River and West Nile viruses, which could be applied to the study of other arboviruses
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