Search

Opportunistic pathogens are environmental microbes that are generally harmless and only occasionally cause disease. Unlike obligate pathogens, the growth and survival of opportunistic pathogens do not rely on host infection or transmission. Their versatile lifestyles make it challenging to decipher how and why virulence has evolved in opportunistic pathogens. The coincidental evolution hypothesis postulates that virulence results from exaptation or pleiotropy, i.e. traits evolved for adaptation to living in one environment that have a different function in another. In particular, adaptation to avoid or survive protist predation has been suggested to contribute to the evolution of bacterial virulence (the training ground hypothesis). Here, we used experimental evolution to determine how the selective pressure imposed by a protist predator impacts the virulence and fitness of a ubiquitous environmental opportunistic bacterial pathogen that has acquired multidrug resistance: Serratia marcescens. To this aim, we evolved S. marcescens in the presence or absence of generalist protist predator, Tetrahymena thermophila. After 60 d of evolution, we evaluated genotypic and phenotypic changes by comparing evolved S. marcescens with the ancestral strain. Whole-genome shotgun sequencing of the entire evolved populations and individual isolates revealed numerous cases of parallel evolution, many more than statistically expected by chance, in genes associated with virulence. Our phenotypic assays suggested that evolution in the presence of a predator maintained virulence, whereas evolution in the absence of a predator resulted in attenuated virulence. We also found a significant correlation between virulence, biofilm formation, growth, and grazing resistance. Overall, our results provide evidence that bacterial virulence and virulence-related traits are maintained by selective pressures imposed by protist predation. [ABSTRACT FROM AUTHOR]

As Wolbachia pipientis is more widely being released into field populations of Aedes aegypti for disease control, the ability to select the appropriate strain for differing environments is increasingly important. A previous study revealed that longer-term quiescence in the egg phase reduced the fertility of mosquitoes, especially those harboring the wAlbB Wolbachia strain. This infertility was also associated with a greater biting rate. Here, we attempt to quantify the effect of this heightened biting behavior on the transmission potential of the dengue virus using a combination of assays for fitness, probing behavior, and vector competence, allowing repeat feeding, and incorporate these effects in a model of R0. We show that Wolbachia-infected infertile mosquitoes are more interested in feeding almost immediately after an initial blood meal relative to wild type and Wolbachia-infected fertile mosquitoes and that these differences continue for up to 8 days over the period we measured. As a result, the infertile Wolbachia mosquitoes have higher virus prevalence and loads than Wolbachia-fertile mosquitoes. We saw limited evidence of Wolbachia-mediated blocking in the disseminated tissue (legs) in terms of prevalence but did see reduced viral loads. Using a previously published estimate of the extrinsic incubation period, we demonstrate that the effect of repeat feeding/infertility is insufficient to overcome the effects of Wolbachia-mediated blocking on R0. These estimates are very conservative, however, and we posit that future studies should empirically measure EIP under a repeat feeding model. Our findings echo previous work where periods of extensive egg quiescence affected the reproductive success of Wolbachia-infected Ae. aegypti. Additionally, we show that increased biting behavior in association with this infertility in Wolbachia-infected mosquitoes may drive greater vector competence. These relationships require further exploration, given their ability to affect the success of field releases of Wolbachia for human disease reduction in drier climates where longer egg quiescence periods are expected. Author summary: Wolbachia pipientis is a naturally occurring, maternally inherited insect endosymbiont that was artificially introduced into the mosquito, Aedes aegypti, the dominant vector of dengue, Zika, chikungunya, and Yellow Fever viruses globally. This bacterium is being released into mosquito populations in the field for disease control because Wolbachia reduces the ability of these co-infecting viruses to replicate as well as spread easily through populations due to a form of reproductive manipulation. Previous research has shown that mosquito eggs experiencing long periods of storage make Wolbachia-infected adults more likely to be infertile and exhibit increased biting frequency. Using a combination of behavioral, vector competence, and fitness assays as well as modeling, we show that the increase in biting behavior leads to higher virus prevalence and load in the mosquito. Together, the decreased fertility and the increased virus transmission potential may decrease the efficacy of Wolbachia, particularly in dry climates where mosquitoes likely spend longer in the egg stage before hatching. We highlight a range of future experiments needed to fully ascertain the effect of egg quiescence on virus transmission. [ABSTRACT FROM AUTHOR]

Female Aedes aegypti (Linnaeus) mosquitoes integrate multiple sensory cues to locate human hosts for blood meals. Although male Ae. aegypti swarm around and land on humans in nature to mate, direct evidence of attraction to humans is limited. Male mosquito attraction to human host cues is often undetectable in confined laboratory assays, leading to a misconception that male mosquitoes are not attracted to humans. We used semifield experiments to demonstrate robust attraction of male Ae. aegypti to humans. Human-baited traps captured up to 25% of released males within 15 min, whereas control traps without humans as bait failed to capture males. Rapid attraction to humans was further demonstrated through videography. Males swarmed around and landed on human subjects, with no activity recorded in paired unbaited controls. Finally, we confirm the lack of discernible male attraction to humans in small laboratory cages. Our experiments demonstrate that both male and female Ae. aegypti show attraction to humans, but with clear sex-specific behavioral differences at short-range. Male mosquito attraction to humans is likely to be important for mating success in wild populations and its basis should be further explored. Our results highlight the importance of arena size and assay design for mosquito behavioral research. A better understanding of host cues that attract males could help us to improve mosquito surveillance and control., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)