Characterizing and Quantifying Arbovirus Transmission by Aedes aegypti Using Forced Salivation and Analysis of Bloodmeals.
Characterizing and Quantifying Arbovirus Transmission by Aedes aegypti Using Forced Salivation and Analysis of Bloodmeals.
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DOI：
10.3390/insects12040304
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发表时间：
2021-03-30
期刊：
影响因子：
3
通讯作者：
Foy BD
中科院分区：
文献类型：
--
作者：
Miller MR;Sorensen MR;Markle ED;Clarkson TC;Knight AL;Savran MJ;Foy BD
蚊子传播数百种虫媒病毒。估计从传染性蚊子传播到宿主的病毒滴度的能力至关重要。在这项研究中，我们评估了强迫流涎技术，以估计和比较蚊子传播的寨卡病毒和基孔肯雅病毒的滴度。我们证明，在吸血后对蚊子进行强制流涎可能是估计吸血期间传播的病毒的有效方法。此外，通过比较喂食后血粉和唾液的滴度，我们发现蚊子在人工喂血期间会重新摄入大部分唾液。这项研究的结果为理解和量化虫媒病毒的传播提供了新的信息。虫媒病毒传播研究取决于估计从传染性蚊子传播到宿主的病毒滴度的能力。有几种方法可以估计蚊子唾液中的病毒滴度，包括（1）使用强制流涎（FS），将传染性蚊子的长鼻被迫进入含有介质的毛细管中，以收集和测试其唾液中的病毒，以及（2）在喂血后立即量化吐入宿主组织或人工饲养器中血液中的病毒。我们研究了 FS 和血粉，以估计和比较由蚊媒埃及伊蚊传播的寨卡病毒和基孔肯雅病毒的滴度。与使用胎牛血清 (FBS) 加甘油相比，使用浸油作为 FS 介质更容易从单个蚊子中检测到两种病毒的传染性病毒和病毒基因组，但 FS 介质对阳性样本的病毒定量没有影响。将从未接受过血粉的蚊子个体或群体与之前刚刚喂过血的蚊子进行比较时，FS 病毒滴度是相当的，这表明吸血不会影响 FS。这表明在吸血后对蚊子进行FS可能是估计吸血期间传播的病毒的有效方法。然而，与从喂血后蚊子的 FS 中定量病毒相比，从喂血后人工喂食器中残留的血液中检测病毒大多不成功。相比之下，免疫功能低下的小鼠在被寨卡病毒感染的蚊子喂食后总是会被感染，即使 FS 在喂血后的唾液中没有检测到感染性病毒。由于这种差异，我们测试了以人工献血器为食的个体蚊子摄入的血粉中的病毒，并将其与从 FS 采集的唾液中的病毒以及体内的病毒进行了比较。这些实验显示，与在同一蚊子唾液中检测到的病毒滴度相比，解剖的血粉中的病毒滴度高出约 50-100 倍，这证明了蚊子在人工献血过程中如何重新摄入大部分唾液，并强调了埃及伊蚊献血过程中病毒传播的大幅增加。 FS和人工喂养的蚊子的解剖血粉均表明，蚊子吐出的病毒RNA数量比传染性病毒数量多2-5个对数。这项研究的结果为理解和量化埃及伊蚊虫媒病毒的传播提供了重要信息。
Mosquitoes transmit hundreds of arboviruses. The ability to estimate the titers of viruses transmitted from infectious mosquitoes to a host is critical. In this study, we evaluated forced salivation techniques to estimate and compare titers of Zika virus and chikungunya virus transmitted by the mosquitoes. We demonstrated that performing forced salivation on mosquitoes after blood feeding might be an efficient way to estimate the virus transmitted during blood feeding. Additionally, by comparing titers of bloodmeals and saliva post-feeding, we showed that mosquitoes re-ingest much of their saliva during artificial blood feeding. The results from this study add new information to understanding and quantifying the transmission of arboviruses. Arbovirus transmission studies are dependent on the ability to estimate the titer of virus transmitted from infectious mosquitoes to a host. There are several methods for estimating virus titer in mosquito saliva, including (1) using forced salivation (FS) whereby the infectious mosquito’s proboscis is forced into a capillary tube containing media to collect and test their saliva for virus, and (2) by quantifying virus expectorated into host tissues or into the blood contained in an artificial feeder immediately after blood feeding. We studied FS and bloodmeals to estimate and compare titers of Zika virus and chikungunya virus transmitted by the mosquito vector Aedes aegypti. Infectious virus and viral genomes of both viruses were detected more often from individual mosquitoes using immersion oil for the FS media compared to fetal bovine serum (FBS) plus glycerol, but the FS media had no influence on virus quantification from positive samples. FS virus titers were equivalent when comparing individuals or groups of mosquitoes that never received a blood meal compared to those that were blood fed immediately prior, showing that blood feeding does not influence FS. This suggested that performing FS on mosquitoes after blood feeding might be an efficient way to estimate virus transmitted during blood feeding. However, detecting virus from the blood remaining in an artificial feeder post-blood feeding was mostly unsuccessful relative to quantifying virus from FS of the post-blood fed mosquitoes. In contrast, immunocompromised mice always became infected after being fed on by Zika-infected mosquitoes, even when no infectious virus was detected in their saliva by FS post-blood feed. Due to this discrepancy, we tested the ingested bloodmeals of individual mosquitoes that fed on artificial blood feeders for virus, and compared these to virus in their saliva harvested from FS and to virus in their bodies. These experiments revealed ~50–100 times higher virus titers in the dissected bloodmeals compared to those detected in the same mosquitoes’ saliva, demonstrating how mosquitoes re-ingest much of their saliva during artificial blood feeding, and highlighting a large increase in virus transmission during Aedes aegypti blood feeding. Both FS and the dissected bloodmeals of artificially blood-fed mosquitoes showed that the quantity of viral RNA expectorated by mosquitoes was 2–5 logs more than the quantity of infectious virus. The results from this study add critical information to understanding and quantifying the transmission of Aedes aegypti arboviruses.
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影响因子：
1.9
作者：
Ribeiro, JMC
通讯作者：
Ribeiro, JMC
影响因子：
3.7
作者：
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DOI：
10.4269/ajtmh.1966.15.989
发表时间：
1966-01-01
影响因子：
3.3
作者：
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通讯作者：
HURLBUT, HS
影响因子：
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作者：
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通讯作者：
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DOI：
10.2987/8756-971x(2005)21
发表时间：
2005-09-01
影响因子：
1
作者：
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通讯作者：
Andre, RG