Revisiting Alkali Metals As a Tool to Characterize Patterns of Mosquito Dispersal and Oviposition.

Mark-recapture methods constitute a set of classical ecological tools that are used to collect information on species dispersal and population size. These methods have advanced knowledge in disparate scientific fields, from conservation biology to pest control. Gathering information on the dispersal of mosquito species, such as Aedes aegypti, has become critical since the recognition of their role as vectors of pathogens. Here, we evaluate a method to mark mosquitoes that exploits the rare alkali metals rubidium (Rb) and caesium (Cs), which have been used previously to mark adult insects through feeding. We revised this method by adding Rb and Cs directly to water in which the immature stages of Ae. aegypti were allowed to develop. We then assessed the effect of Rb- and Cs-enriched water on fitness, survival and bioaccumulation in both adult females and their eggs. Results indicated that Cs had adverse effects on Ae. aegypti, even at low concentrations, whereas Rb at low concentrations had no measured effects on exposed individuals and accumulated at detectable levels in adult females. The method described here relies on passive uptake of Rb during immature stages, which has the benefit of avoiding handling or manipulation of the dispersive adults, which enables purer measurement of movement. Moreover, we demonstrated that Rb was transferred efficiently from the marked females to their eggs. To our knowledge, Rb is the only marker used for mosquitoes that has been shown to transfer vertically from females to eggs. The application of Rb rather than more traditional markers may therefore increase the quality (no impact on released individuals) and quantity (both adults and eggs are marked) of data collected during MR studies. The method we propose here can be used in combination with other markers, such as stable isotopes, in order to maximize the information collected during MR experiments

Vector competence of Aedes aegypti, Culex tarsalis, and Culex quinquefasciatus from California for Zika virus.

Zika virus (ZIKV) has emerged since 2013 as a significant global human health threat following outbreaks in the Pacific Islands and rapid spread throughout South and Central America. Severe congenital and neurological sequelae have been linked to ZIKV infections. Assessing the ability of common mosquito species to transmit ZIKV and characterizing variation in mosquito transmission of different ZIKV strains is important for estimating regional outbreak potential and for prioritizing local mosquito control strategies for Aedes and Culex species. In this study, we evaluated the laboratory vector competence of Aedes aegypti, Culex quinquefasciatus, and Culex tarsalis that originated in areas of California where ZIKV cases in travelers since 2015 were frequent. We compared infection, dissemination, and transmission rates by measuring ZIKV RNA levels in cohorts of mosquitoes that ingested blood meals from type I interferon-deficient mice infected with either a Puerto Rican ZIKV strain from 2015 (PR15), a Brazilian ZIKV strain from 2015 (BR15), or an ancestral Asian-lineage Malaysian ZIKV strain from 1966 (MA66). With PR15, Cx. quinquefasciatus was refractory to infection (0%, N = 42) and Cx. tarsalis was infected at 4% (N = 46). No ZIKV RNA was detected in saliva from either Culex species 14 or 21 days post feeding (dpf). In contrast, Ae. aegypti developed infection rates of 85% (PR15; N = 46), 90% (BR15; N = 20), and 81% (MA66; N = 85) 14 or 15 dpf. Although MA66-infected Ae. aegypti showed higher levels of ZIKV RNA in mosquito bodies and legs, transmission rates were not significantly different across virus strains (P = 0.13, Fisher's exact test). To confirm infectivity and measure the transmitted ZIKV dose, we enumerated infectious ZIKV in Ae. aegypti saliva using Vero cell plaque assays. The expectorated plaque forming units PFU varied by viral strain: MA66-infected expectorated 13±4 PFU (mean±SE, N = 13) compared to 29±6 PFU for PR15-infected (N = 13) and 35±8 PFU for BR15-infected (N = 6; ANOVA, df = 2, F = 3.8, P = 0.035). These laboratory vector competence results support an emerging consensus that Cx. tarsalis and Cx. quinquefasciatus are not vectors of ZIKV. These results also indicate that Ae. aegypti from California are efficient laboratory vectors of ancestral and contemporary Asian lineage ZIKV

Revisiting Alkali Metals As a Tool to Characterize Patterns of Mosquito Dispersal and Oviposition

Mark-recapture methods constitute a set of classical ecological tools that are used to collect information on species dispersal and population size. These methods have advanced knowledge in disparate scientific fields, from conservation biology to pest control. Gathering information on the dispersal of mosquito species, such as Aedes aegypti, has become critical since the recognition of their role as vectors of pathogens. Here, we evaluate a method to mark mosquitoes that exploits the rare alkali metals rubidium (Rb) and caesium (Cs), which have been used previously to mark adult insects through feeding. We revised this method by adding Rb and Cs directly to water in which the immature stages of Ae. aegypti were allowed to develop. We then assessed the effect of Rb- and Cs-enriched water on fitness, survival and bioaccumulation in both adult females and their eggs. Results indicated that Cs had adverse effects on Ae. aegypti, even at low concentrations, whereas Rb at low concentrations had no measured effects on exposed individuals and accumulated at detectable levels in adult females. The method described here relies on passive uptake of Rb during immature stages, which has the benefit of avoiding handling or manipulation of the dispersive adults, which enables purer measurement of movement. Moreover, we demonstrated that Rb was transferred efficiently from the marked females to their eggs. To our knowledge, Rb is the only marker used for mosquitoes that has been shown to transfer vertically from females to eggs. The application of Rb rather than more traditional markers may therefore increase the quality (no impact on released individuals) and quantity (both adults and eggs are marked) of data collected during MR studies. The method we propose here can be used in combination with other markers, such as stable isotopes, in order to maximize the information collected during MR experiments

Impact of temperature on the extrinsic incubation period of Zika virus in Aedes aegypti.

Since Zika virus (ZIKV) emerged as a global human health threat, numerous studies have pointed to Aedes aegypti as the primary vector due to its high competence and propensity to feed on humans. The majority of vector competence studies have been conducted between 26-28°C, but arboviral extrinsic incubation periods (EIPs), and therefore transmission efficiency, are known to be affected strongly by temperature. To better understand the relationship between ZIKV EIPs and temperature, we evaluated the effect of adult mosquito exposure temperature on ZIKV infection, dissemination, and transmission in Ae. aegypti at four temperatures: 18°C, 21°C, 26°C, and 30°C. Mosquitoes were exposed to viremic mice infected with a 2015 Puerto Rican ZIKV strain, and engorged mosquitoes were sorted into the four temperatures with 80% RH and constant access to 10% sucrose. ZIKV infection, dissemination, and transmission rates were assessed via RT-qPCR from individual mosquito bodies, legs and wings, and saliva, respectively, at three to five time points per temperature from three to 31 days, based on expectations from other flavivirus EIPs. The median time from ZIKV ingestion to transmission (median EIP, EIP50) at each temperature was estimated by fitting a generalized linear mixed model for each temperature. EIP50 ranged from 5.1 days at 30°C to 24.2 days at 21°C. At 26°C, EIP50 was 9.6 days. At 18°C, only 15% transmitted by day 31 so EIP50 could not be estimated. This is among the first studies to characterize the effects of temperature on ZIKV EIP in Ae. aegypti, and the first to do so based on feeding of mosquitoes on a live, viremic host. This information is critical for modeling ZIKV transmission dynamics to understand geographic and seasonal limits of ZIKV risk; it is especially relevant for determining risk in subtropical regions with established Ae. aegypti populations and relatively high rates of return travel from the tropics (e.g. California or Florida), as these regions typically experience cooler temperature ranges than tropical regions

ZIKV infection, dissemination, and transmission rates in California <i>Aedes</i> and <i>Culex</i> mosquitoes 14 and 21 days post ingestion of blood from viremic mice.

<p>ZIKV infection, dissemination, and transmission rates in California <i>Aedes</i> and <i>Culex</i> mosquitoes 14 and 21 days post ingestion of blood from viremic mice.</p

Infecting (bodies), disseminating (legs+wings) and transmitted (saliva) ZIKV RNA levels 14 or 15 days after <i>Ae</i>. <i>aegypti</i> orally ingested one of three Asian lineage ZIKV strains.

<p>Each dot represents the mean log₁₀ ZIKV genome copies per tissue or saliva sample from an individual <i>Ae</i>. <i>aegypti</i>. <i>Ae</i>. <i>aegypti</i> from Los Angeles, California, USA, were fed on viremic mice infected with ZIKV from Malaysia 1966 (MA66), Puerto Rico 2015 (PR15) or Brazil 2015 (BR15). Mosquitoes exposed to BR15 were assayed 15 dpf, MA66 and PR15 cohorts were assayed 14 dpf. Each dot represents the mean of two or more RT-qPCR technical replicates. The dashed lines represent the limits of detection (LOD). Dots below dashed line represent tested samples with an undetectable Ct or a Ct value of >38. The LOD for saliva was lower than for tissues because RNA was extracted from a higher proportion of the saliva sample volume. Asterisks show significant differences in means across groups of the same tissue type, <i>P</i><0.01, ANOVA, Tukey post-hoc test. No asterisk indicates no significant difference across groups.</p

Individual mosquito ZIKV RNA levels in body, legs+wings, and saliva of <i>Aedes aegypti</i> from Los Angeles, CA, USA.

<p><i>Ae</i>. <i>aegypti</i> from Los Angeles ingested blood from viremic ZIKV-infected interferon receptor deficient mice that had been inoculated with different ZIKV strains. Each colored box represents an individual mosquito sample showing the magnitude of ZIKV RNA detected in each body (B), legs+wings (LW), and saliva (S). The red to blue color scale shows high (red) to low (blue) ZIKV RNA levels. Samples with no detectable ZIKV RNA are colored white. ZIKV BR15-infected mosquitoes were not tested 21 dpf.</p

The influence of vector-borne disease on human history: socio-ecological mechanisms

Vector-borne diseases (VBDs) are embedded within complex socio-ecological systems. While research has traditionally focused on the direct effects of VBDs on human morbidity and mortality, it is increasingly clear that their impacts are much more pervasive. VBDs are dynamically linked to feedbacks between environmental conditions, vector ecology, disease burden, and societal responses that drive transmission. As a result, VBDs have had profound influence on human history. Mechanisms include: (1) killing or debilitating large numbers of people, with demographic and population-level impacts; (2) differentially affecting populations based on prior history of disease exposure, immunity, and resistance; (3) being weaponised to promote or justify hierarchies of power, colonialism, racism, classism and sexism; (4) catalysing changes in ideas, institutions, infrastructure, technologies and social practices in efforts to control disease outbreaks; and (5) changing human relationships with the land and environment. We use historical and archaeological evidence interpreted through an ecological lens to illustrate how VBDs have shaped society and culture, focusing on case studies from four pertinent VBDs: plague, malaria, yellow fever and trypanosomiasis. By comparing across diseases, time periods and geographies, we highlight the enormous scope and variety of mechanisms by which VBDs have influenced human history