EHDV-2 Infection Prevalence Varies in Culicoides sonorensis after Feeding on Infected White-Tailed Deer over the Course of Viremia

Epizootic hemorrhagic disease viruses (EHDVs) are arboviral pathogens of white-tailed deer and other wild and domestic ruminants in North America. Transmitted by various species of Culicoides, EHDVs circulate wherever competent vectors and susceptible ruminant host populations co-exist. The impact of variation in the level and duration of EHDV viremia in white-tailed deer (Odocoileus virginianus) on Culicoides infection prevalence is not well characterized. Here we examined how infection prevalence in a confirmed North American vector of EHDV-2 (Culicoides sonorensis) varies in response to fluctuations in deer viremia. To accomplish this, five white-tailed deer were experimentally infected with EHDV-2 and colonized C. sonorensis were allowed to feed on deer at 3, 5, 7, 10, 12, 14, 18, and 24 days post infection (dpi). Viremia profiles in deer were determined by virus isolation and titration at the same time points. Blood-fed Culicoides were assayed for virus after a 10-day incubation (27 ◦C) period. We found that increases in deer EHDV blood titers significantly increased both the likelihood that midges would successfully acquire EHDV and the proportion of midges that reached the titer threshold for transmission competence. Unexpectedly, we identified four infected midge samples (three individuals and one pool) after feeding on one deer 18 and 24 dpi, when viremia was no longer detectable by virus isolation. The ability of ruminants with low-titer viremia to serve as a source of EHDV for blood-feeding Culicoides should be explored further to better understand its potential epidemiological significance

Data from: Rate of novel host invasion affects adaptability of evolving RNA virus lineages

Although differing rates of environmental turnover should be consequential for the dynamics of adaptive change, this idea has been rarely examined outside of theory. In particular, the importance of RNA viruses in disease emergence warrants experiments testing how differing rates of novel host invasion may impact the ability of viruses to adaptively shift onto a novel host. To test whether the rate of environmental turnover influences adaptation, we experimentally evolved 144 Sindbis virus lineages in replicated tissue-culture environments, which transitioned from being dominated by a permissive host cell type to a novel host cell type. The rate at which the novel host ‘invaded’ the environment varied by treatment. The fitness (growth rate) of evolved virus populations was measured on each host type, and molecular substitutions were mapped via whole genome consensus sequencing. Results showed that virus populations more consistently reached high fitness levels on the novel host when the novel host ‘invaded’ the environment more gradually, and gradual invasion resulted in less variable genomic outcomes. Moreover, virus populations that experienced a rapid shift onto the novel host converged upon different genotypes than populations that experienced a gradual shift onto the novel host, suggesting a strong effect of historical contingency

Fitness_Assay_Data

Fitness data for evolved Sindbis virus populations. Fitness is measured as viral titer (pfu/mL) on each cell type (BHK and CHO). Each virus population was assayed in triplicate on each host type. The ancestral Sindbis virus used to found experimental Sindbis lineages was assayed nine times on each host

Fitness_Assay_Data

Fitness data for evolved Sindbis virus populations. Fitness is measured as viral titer (pfu/mL) on each cell type (BHK and CHO). Each virus population was assayed in triplicate on each host type. The ancestral Sindbis virus used to found experimental Sindbis lineages was assayed nine times on each host

EHDV-2 Infection Prevalence Varies in <i>Culicoides sonorensis</i> after Feeding on Infected White-Tailed Deer over the Course of Viremia

Epizootic hemorrhagic disease viruses (EHDVs) are arboviral pathogens of white-tailed deer and other wild and domestic ruminants in North America. Transmitted by various species of Culicoides, EHDVs circulate wherever competent vectors and susceptible ruminant host populations co-exist. The impact of variation in the level and duration of EHDV viremia in white-tailed deer (Odocoileus virginianus) on Culicoides infection prevalence is not well characterized. Here we examined how infection prevalence in a confirmed North American vector of EHDV-2 (Culicoides sonorensis) varies in response to fluctuations in deer viremia. To accomplish this, five white-tailed deer were experimentally infected with EHDV-2 and colonized C. sonorensis were allowed to feed on deer at 3, 5, 7, 10, 12, 14, 18, and 24 days post infection (dpi). Viremia profiles in deer were determined by virus isolation and titration at the same time points. Blood-fed Culicoides were assayed for virus after a 10-day incubation (27 &#176;C) period. We found that increases in deer EHDV blood titers significantly increased both the likelihood that midges would successfully acquire EHDV and the proportion of midges that reached the titer threshold for transmission competence. Unexpectedly, we identified four infected midge samples (three individuals and one pool) after feeding on one deer 18 and 24 dpi, when viremia was no longer detectable by virus isolation. The ability of ruminants with low-titer viremia to serve as a source of EHDV for blood-feeding Culicoides should be explored further to better understand its potential epidemiological significance

Sample preparation techniques for GC

Sample preparation is still considered the most time-consuming and error-prone step within the analytical process in many research fields. This is particularly true in food and environmental analysis where the complexity of many of the investigated matrices and the low concentration levels at which the target compounds should be accurately determined made necessary the use of tedious and highly manipulative multistep sample preparation protocols. This chapter reviews current state of the art in the field of sample preparation for combined use with gas chromatographic-based techniques. The most relevant developments achieved in the last two decades in this active research area have been reviewed and discussed on the basis of representative application studies primarily taken from the environmental and food fields. As in other research areas, miniaturisation and increased integration of the several treatment steps typically required for the preparation of these matrices are revealed as the most relevant trends within this step of the analytical process.Author thanks MINECO (CTQ2010-32957) and CM and FEDER program (project S2009/AGR-1464, ANALISYC-II) for financial support.Peer reviewe