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Short, S. M.

Publications and source records attributed to Short, S. M..

13 recordsLinked to original sources

The microbiota impacts life history traits and mating success in male Aedes aegypti mosquitoes

Aedes aegypti mosquitoes transmit multiple arboviruses, but population suppression through the mass release of sterile or incompatible male mosquitoes can effectively reduce populations. These methods depend on the reliable mass-rearing of healthy, robust males that can successfully mate with wild females. The microbiota, a critical component of the larval diet, can dramatically influence life history traits relevant to mass-rearing and male quality. Here, we used axenic (microbe-free), monoxenic (inoculated with E. coli), and "laboratory community" mosquitoes (inoculated with an undefined microbiota derived from laboratory mosquitoes) to show that longevity was significantly enhanced in axenic and monoxenic males compared to laboratory community males. Moreover, monoxenic males more efficiently obtained mates in non-competitive mating scenarios compared to laboratory community males. However, microbiota treatment had no effect when males from different treatments competed for a mate. Our findings suggest that the microbiota is a key determinant of male mosquito life history with direct implications for optimizing production of males for control programs. TeaserThe microbiota of Aedes aegypti mosquitoes affects multiple male life history traits.

microbiology↗

The Aedes aegypti bacterial microbiota is robust to infection with the obligate microsporidian parasite Edhazardia aedis.

Edhazardia aedis is an obligate microsporidian parasite of the arthropod vector Aedes aegypti, which is responsible for the spread of several vertebrate pathogens of global health importance. E. aedis can be highly virulent to Ae. aegypti and infection has severely detrimental effects on multiple life history traits that are relevant to the vectoral capacity of Ae. aegypti, including longevity, body size, propensity to host-seek and blood-feed, and reproductive capacity. Because E. aedis is also highly specific to Ae. aegypti and is incapable of completing its full life cycle in any other mosquito species, E. aedis merits investigation as a novel tool for biological vector control. In the present study, we queried the effect of E. aedis infection on the bacterial microbiota of adult female Ae. aegypti using high-throughput amplicon sequencing of the 16S rRNA gene. Analysis of sequencing data revealed that the bacterial microbiota community is strikingly robust to E. aedis infection, as we observed no significant effect on alpha or beta diversity, differential abundance of any taxa, predicted metabolic function profile, or overall bacterial load. The data show that E. aedis, despite dramatically impacting the health and fitness of the adult female mosquito, does not affect the microbiota. These results provide unique insight into tripartite relationships (or lack thereof) between hosts, pathogens, and the microbiota.

immunology↗

Addition of a single bacterial isolate to conventional larval rearing water can impact wing size and longevity in adult male Aedes aegypti.

Male mosquitoes are mass-reared around the globe for use in mosquito control programs like Sterile Insect Technique and Incompatible Insect Technique. During larval development, mosquitoes co-exist with complex microbial communities that serve as food and also form the internal microbiota of the organism. The microbiota can impact multiple larval and adult life history traits including development rate and male body size. In the present study, we investigated how male wing length and adult male longevity is impacted by the addition of a single bacterial isolate to otherwise conventional larval rearing water. Of three isolates tested, we found that larval exposure to one, Cedecea sp., resulted in adult males with significantly reduced wing length and longevity. Our findings suggest that minor modifications of the microbial community during larval development can have life-long effects on mosquitoes which, like many organisms, acquire their microbiota from the environment. Moreover, they suggest that mosquito life history traits can be influenced by small shifts in the microbial rearing community, which could impact the efficacy and efficiency of mosquito mass rearing for vector control.

microbiology↗

A polinton-like virus of the algae Chrysochromulina parva inhibits the growth of a newly isolated relative of Tethysvirus ontarioense

Previous discovery of genomes of polinton-like viruses (PLVs) associated with viruses of Chrysochromulina parva stimulated this research to determine the biological nature of these putative viral hyperparasites. Purification of C. parva viruses to enable co-infection experiments led to the discovery of a previously unknown virus, CpV-BQ3, which, based on sequence information and electron microscopy, is a species of Tethysvirus, a genus within the Megaviricetes. Purification and TEM imaging of CpV-PLV Moe revealed naked icosahedral particles morphologically similar other cultivated virophages and PLVs. Mixed-infection experiments with the putative polinton-like virus CpV-PLV Moe demonstrated that CpV-BQ3 supports its replication whereas the putative Phycodnavirus CpV-BQ1 does not. Further, experimental infections with differing proportions of the Moe and its helper virus CpV-BQ3 revealed a dose-effect whereby high levels of Moe had a greater negative impact on BQ3 replication compared to lower levels. Conversely, high levels of Moe relative to BQ3 provided greater protection for C. parva allowing enhanced cell survival whereas low doses of Moe did not prevent cell lysis. Overall, the results of this study demonstrated the intimate relationship of CpV-PLV Moe with the newly discovered virus, CpV-BQ3, and C. parva, and illustrate the complex ecology of algal viruses.

microbiology↗

Hybrid sequencing reveals the genome of a Chrysochromulina parva virus and highlight its distinct replication strategy

Chrysochromulina parva (C. parva) is a eukaryotic freshwater haptophyte algae found in lakes and rivers worldwide. It is known to be infected by viruses, yet knowledge of the diversity and activity of these viruses is still very limited. Based on sequences of PCR-amplified polB gene fragments, Chrysochromulina parva virus BQ1 (CpV-BQ1) was the first known lytic agent of C. parva, and was considered a member of the virus family Phycodnaviridae, order Algavirales. However, the genome of a different C. parva-infecting virus (CpV-BQ2, or Tethysvirus ontarioense) from another virus family, the Mesomimiviridae, order Imitervirales, was the first sequenced. Here, we report the complete genome sequence of the putative phycodnavirus CpV-BQ1, accession PQ783904. The complete CpV-BQ1 genome sequence is 165,454 bp with a GC content of 32.32% and it encodes 193 open reading frames. Phylogenetic analyses of several virus hallmark genes including the DNA polymerase (polB), the late gene transcription factor (VLTF-3), and the putative A32-like virion packaging ATPase (Viral A32) all demonstrate that CpV-BQ1 is most closely related to other viruses in the phylum Megaviricetes within the order Algavirales, family Phycodnaviridae.

genomics↗

Establishment and comparative genomics of a high-quality collection of mosquito-associated bacterial isolates -- MosAIC (Mosquito-Associated Isolate Collection)

Mosquitoes transmit medically important human pathogens, including viruses like dengue virus and parasites such as Plasmodium spp., the causative agent of malaria. Mosquito microbiomes are critically important for the ability of mosquitoes to transmit disease-causing agents. However, while large collections of bacterial isolates and genomic data exist for vertebrate microbiomes, the vast majority of work in mosquitoes to date is based on 16S rRNA gene amplicon data that provides limited taxonomic resolution and no functional information. To address this gap and facilitate future studies using experimental microbiome manipulations, we generated a bacterial Mosquito-Associated Isolate Collection (MosAIC) consisting of 392 bacterial isolates with extensive metadata and high-quality draft genome assemblies that are publicly available for use by the scientific community. MosAIC encompasses 142 species spanning 29 bacterial families, with members of the Enterobacteriaceae comprising 40% of the collection. Phylogenomic analysis of three genera, Enterobacter, Serratia, and Elizabethkingia, reveal lineages of mosquito-associated bacteria isolated from different mosquito species in multiple laboratories. Investigation into species pangenomes further reveals clusters of genes specific to these lineages, which are of interest for future work to identify functions underlying mosquito host association. Altogether, we describe the generation of a physical collection of mosquito-associated bacterial isolates, their genomic data, and analyses of selected groups in context of genome data from closely related isolates, providing a unique, highly valuable resource to investigate factors for bacterial colonisation and adaptation within mosquito hosts. Future efforts will expand the collection to include broader geographic and host species representation, especially from individuals collected from field populations, as well as other mosquito-associated microbes, including fungi, archaea, and protozoa.

microbiology↗

The microbiota of Amblyomma americanum reflects known westward expansion.

AbstractAmblyomma americanum, a known vector of multiple tick-borne pathogens, has expanded its geographic distribution across the United States in the past decades. Tick microbiomes may play a role shaping their hosts life history and vectorial capacity. Bacterial communities associated with A. americanum may reflect, or enable, geographic expansion and studying the microbiota will improve understanding of tick- borne disease ecology. We examined the microbiota structure of 189 adult ticks collected in four regions encompassing their historical and current geographic distribution. Both geographic region of origin and sex were significant predictors of alpha diversity. As in other tick models, within-sample diversity was low and uneven given the presence of dominant endosymbionts. Beta diversity analyses revealed that bacterial profiles of ticks of both sexes collected in the West were significantly different from those of the Historic range. Biomarkers were identified for all regions except the historical range. In addition, Bray-Curtis dissimilarities overall increased with distance between sites. Relative quantification of ecological processes showed that, for females and males, respectively, drift and dispersal limitation were the primary drivers of community assembly. Collectively, our findings highlight how microbiota structural variance discriminates the western-expanded populations of A. americanum ticks from the Historical range. Spatial autocorrelation, and particularly the detection of non- selective ecological processes, are indicative of geographic isolation. Our conclusions demonstrate the value of synergistic analysis of biogeographic and microbial ecology data in investigating range expansion in A. americanum and potentially other tick vectors as well. ImportanceThe incidence of tick-borne diseases is on the rise worldwide, including in the United States. This increase in cases is in large part due to the geographic expansion of tick vectors. Among them is the Lone Star tick, Amblyomma americanum, and understanding the factors driving its recent expansion is important for risk assessment and tick population control. Climate change and human activities have previously been posited to be main driving forces for their expansion. New sources of relevant biological data can further our understanding of this phenomenon. Microbial communities associated with ticks include bacteria that affect pathogen transmission and tick behaviors. Here, we investigated whether bacterial community and geographic location data could be combined to better understand the expansion of A. americanum populations. Its not clear whether differences in the microbiota are reflective of or facilitating expansion, but our findings suggest this approach is promising and warrants further investigation.

ecology↗

Temporally dynamic effects of adult diet on resistance and tolerance to bacterial infection in Aedes aegypti

Immune defense is comprised of 1) resistance: the ability to reduce pathogen load, and 2) tolerance: the ability to limit the disease severity induced by a given pathogen load. The study of tolerance in the field of animal immunity is fairly nascent in comparison to resistance. Consequently, studies which examine immune defense comprehensively (i.e., considering both resistance and tolerance in conjunction) are uncommon, despite their exigency in achieving a thorough understanding of immune defense. Furthermore, understanding tolerance in arthropod disease vectors is uniquely relevant, as tolerance is essential to the cyclical transmission of pathogens by arthropods. Here, we tested the effect(s) of dietary sucrose concentration (high or low) and blood meal (present or absent) on resistance and tolerance to Escherichia coli infection in the yellow fever mosquito Aedes aegypti. Resistance and tolerance were measured concurrently and at multiple timepoints. We found that both blood and sucrose affected resistance. Mosquitoes from the low sugar treatment displayed enhanced resistance at all timepoints post-infection compared to those from the high sugar treatment. Additionally, blood-fed mosquitoes showed enhanced resistance compared to non-blood-fed mosquitoes, but only on day 1 post-infection. Sucrose had no effect on tolerance, but the effect of blood was significant and dynamic across time. Specifically, we show that consuming blood prior to infection ameliorates a temporal decline in tolerance that mosquitoes experience when provided with only sugar meals. Taken together, our findings indicate that different dietary components can have unique and sometimes temporally dynamic impacts on resistance and tolerance. Finally, our findings 1) highlight the value of experimental and analytical frameworks which consider the explicit testing of effects on both resistance and tolerance as separate, but equally important, components of immune defense, and 2) underscore the importance of including a temporal component in studies of immune defense.

immunology↗

Analysis of prokaryotic genes in an impaired Great Lakes harbour reveals seasonal metabolic shifts and a previously undetected cyanobacterium

As primary drivers of organic matter re-mineralization and trophic carbon transfer, bacteria promote biogeochemical cycling and shape ecosystems globally. We studied bacterial communities spatially and seasonally in an impaired harbour of Lake Ontario by extracting and sequencing community DNA from water samples collected biweekly from different sites. Assembled contigs were annotated at the phylum level, and Cyanobacteria were further characterized at order and species levels. Actinobacteria were most abundant in early summer, while Cyanobacteria were dominant in mid-summer. Microcystis aeruginosa and Limnoraphis robusta were most abundant throughout the sampling period, expanding the documented diversity of Cyanobacteria in Hamilton Harbour. Functional annotations were performed using the MG-RAST pipeline and SEED database, revealing that genes for photosynthesis, nitrogen metabolism and aromatic compound metabolism varied in relative abundances over the season, while phosphorus metabolism remained consistent, suggesting that these genes remain essential despite fluctuating environmental conditions and community succession. We observed seasonal shifts from anoxygenic to oxygenic phototrophy, and from ammonia assimilation to nitrogen fixation, coupled with decreasing heterotrophic bacteria and increasing cyanobacteria relative abundances. Our data contribute important insights into bacterial taxa and functional potentials in Hamilton Harbour, revealing seasonal and spatial dynamics that can be used to inform ongoing remediation efforts.

microbiology↗

Bacterial communities of Aedes aegypti mosquitoes differ between crop and midgut tissues

Microbiota studies of Aedes aegypti and other mosquitoes generally focus on the bacterial communities found in adult female midguts. However, other compartments of the digestive tract maintain communities of bacteria which remain almost entirely unstudied. For example, the Dipteran crop stores nectar and other sugars, but few studies have looked at the microbiome of crops in mosquitoes, and only a single previous study has investigated the crop in Ae. aegypti. In this study, we used both culture-dependent and culture-independent methods to compare the bacterial communities in midguts and crops of laboratory-reared Ae. aegypti. Both methods revealed a trend towards higher abundance, but also higher variability, of bacteria in the midgut than the crop. When present, bacteria from the genus Elizabethkingia (family Weeksellaceae) dominated midgut bacterial communities. In crops, we found a higher diversity of bacteria, and these communities were generally dominated by acetic acid bacteria (family Acetobacteriaceae) from the genera Tanticharoenia and Asaia. These three taxa drove significant community structure differences between the tissues. We used FAPROTAX to predict the metabolic functions of these communities and found that crop bacterial communities were significantly more likely to contain bacteria capable of methanol oxidation and methylotrophy. Both the presence of acetic acid bacteria (which commonly catabolize sugar to produce acetic acid) and the functional profile that includes methanol oxidation (which is correlated with bacteria found with natural sources like nectar) may relate to the presence of sugar in the crop. A better understanding of what bacteria are present in the digestive tract of mosquitoes and how these communities assemble will inform how the microbiota impacts mosquito physiology and the full spectrum of functions provided by the microbiota. It may also facilitate better methods of engineering the mosquito microbiome for vector control or prevention of disease transmission. Author summaryBacteria inside mosquitoes guts have been found to have an impact on mosquito life history traits (such as longevity and fecundity) as well as their susceptibility to infection by human pathogens. Engineering these communities may provide an effective and safe way to control mosquitoes and reduce the impact of the pathogens they spread. In this work, we assayed the bacteria found in midgut and crop tissues of a medically important mosquito, Aedes aegypti. Our results show that these tissues harbor communities of bacteria that differ in composition and function and vary in abundance. Experiments like ours are important to better understand where bacteria are found in an insects body and how these communities assemble. This knowledge may help future researchers more successfully engineer bacterial communities in mosquitoes.

microbiology↗

Comparing the diversity and relative abundance of free and particle-associated aquatic viruses

Metagenomics has enabled rapid increases in virus discovery, in turn permitting revisions of viral taxonomy and our understanding of the ecology of viruses and their hosts. Inspired by recent discoveries of large viruses prevalent in the environment, we re-assessed the longstanding approach of filtering water through small pore-size filters to separate viruses from cells before sequencing. We studied assembled contigs derived from < 0.45 m and > 0.45 m size fractions that were annotated as viral to determine the diversity and relative abundances of virus groups from each fraction. Virus communities were vastly different when comparing the size fractions, indicating that analysis of either fraction alone would provide only a partial perspective of environmental viruses. At the level of virus order/family we observed highly diverse and distinct virus communities in the > 0.45 m size fractions, whereas the < 0.45 m size fractions were comprised primarily of highly diverse Caudovirales. The relative abundances of Caudovirales for which hosts could be inferred varied widely between size fractions with higher relative abundances of cyanophages in the > 0.45 m size fractions potentially indicating replication within cells during ongoing infections. Many of the Mimiviridae and Phycodnaviridae, and all Iridoviridae and Poxviridae were detected exclusively in the often disregarded > 0.45 m size fractions. In addition to observing unique virus communities associated with each size fraction, we detected viruses common to both fractions and argue that these are candidates for further exploration because they may be the product of ongoing or recent lytic events. IMPORTANCEMost studies of aquatic virus communities analyze DNA sequences derived from the smaller, "free virus" size fraction. Our study demonstrates that analysis of virus communities using only the smaller size fraction can lead to erroneously low diversity estimates for many of the larger viruses such as Mimiviridae, Phycodnaviridae, Iridoviridae, and Poxviridae, whereas analyzing only the larger, > 0.45 m size fraction can lead to underestimates of Caudovirales diversity and relative abundance. Similarly, our data shows that examining only the smaller size fraction can lead to underestimation of virophage and cyanophage relative abundances that could, in turn, cause researchers to assume their limited ecological importance. Given the considerable differences we observed in this study, we recommend cautious interpretations of environmental virus community assemblages and dynamics when based on metagenomic data derived from different size fractions.

microbiology↗

Metagenomic analysis of virus diversity and relative abundance in a eutrophic freshwater harbour

Aquatic viruses have been extensively studied over the past decade, yet fundamental aspects of freshwater virus communities remain poorly described. Our goal was to characterize particle-associated virus communities seasonally and spatially in a freshwater harbour. Community DNA was extracted from water samples and sequenced on an Illumina HiSeq platform. Assembled contigs were annotated as belonging to the virus families Caudovirales, Mimiviridae, Phycodnaviridae, and virophages (Lavidaviridae), or to other groups of undefined viruses. Diverse Mimiviridae contigs were detected in the samples, but the two sites contained distinct Mimiviridae communities. Virophages were often the most abundant group, and discrete virophage taxa were remarkably stable across sites and dates despite fluctuations in Mimiviridae community composition. Caudovirales were present at low abundances in most samples, contrasting other studies of freshwater environments. Similarly, Phycodnaviridae abundances were surprisingly low in all samples despite the harbours capacity to support high algal biomass during the summer and autumn months, suggesting that Mimiviridae are the dominant algae-infecting viruses in this system. Overall, our findings provided insights into freshwater virus community assemblages by expanding the documented diversity of freshwater virus communities, highlighting the potential ecological importance of virophages, and revealing distinct communities over small spatial scales.

microbiology↗

Temporal dynamics of inhalation-linked activity across defined subpopulations of mouse olfactory bulb neurons imaged in vivo.

In mammalian olfaction, inhalation drives the temporal patterning of neural activity that underlies early olfactory processing, and a single inhalation of odorant is sufficient for odor perception. However, how the neural circuits that process incoming olfactory information are activated in the context of inhalation-linked dynamics remains poorly understood. To better understand early olfactory processing in vivo, we used an artificial inhalation paradigm combined with two-photon calcium imaging to compare the dynamics of activity evoked by odorant inhalation across major cell types of the mouse olfactory bulb. Transgenic models and cell-type specific genetic tools were used to express GCaMP6f or jRGECO1a in mitral and tufted cell subpopulations, olfactory sensory neurons and two major juxtaglomerular interneuron classes, and responses to a single inhalation of odorant were compared. Activity in all cell types was strongly linked to inhalation, and all cell types showed some variance in the latency, rise-times and durations of their inhalation-linked response patterns. The dynamics of juxtaglomerular interneuron activity closely matched that of sensory neuron inputs, while mitral and tufted cells showed the highest diversity in dynamics, with a range of latencies and durations that could not be accounted for by heterogeneity in the dynamics of sensory input. Surprisingly, temporal response patterns of mitral and superficial tufted cells were highly overlapping such that these two subpopulations could not be distinguished on the basis of their inhalation-linked dynamics, with the exception of a subpopulation of superficial tufted cells expressing the peptide transmitter cholecystokinin. Overall, these results support a model in which diversity in inhalation-linked patterning of OB output arises first at the level of OSN inputs to the OB and is enhanced by feedforward inhibition from juxtaglomerular interneurons which differentially impacts different subpopulations of OB output neurons.

neuroscience↗