Multiplexed PCR amplicon sequencing (AmpSeq) is an increasingly popular application for cost-effective monitoring of threatened species and managed wildlife populations, and shows strong potential for the genomic epidemiology of infectious disease.AmpSeq data from infectious microbes can inform disease control in multiple ways, such as by measuring drug resistance marker prevalence, distinguishing imported from local cases, and determining the effectiveness of therapeutics. We describe the design and comparative evaluation of two new AmpSeq assays for Plasmodium falciparum malaria parasites: a four-locus panel ("4CAST") composed of highly diverse antigens, and a 129-locus panel ("AMPLseq") composed of drug resistance markers, highly diverse loci for inferring relatedness, and a locus to detect Plasmodium vivax
Anopheles mosquito microbiomes are intriguing ecological niches. Within the gut, microbes adapt to oxidative stress due to heme and iron after blood meals. Although metagenomic sequencing has illuminated spatial and temporal fluxes of microbiome populations, limited data exist on microbial growth dynamics. Here, we analyze growth interactions between a dominant microbiome species, Elizabethkingia anophelis, and other Anopheles‐associated bacteria. We find E. anophelis inhibits a Pseudomonas sp. via an antimicrobial‐independent mechanism and observe biliverdins, heme degradation products, upregulated in cocultures. Purification and characterization of E. anophelis HemS demonstrates heme degradation, and we observe hemS expression is upregulated when cocultured with Pseudomonas sp. This study reveals a competitive microbial interaction between mosquito‐associated bacteria and characterizes the stimulation of heme degradation in E. anophelis when grown with Pseudomonas sp.
Redox
signaling molecules include a number of reactive oxygen species
(ROS), reactive nitrogen species (RNS), and reactive sulfur species
(RSS). These molecules work collectively in the regulation of many
physiological processes. Understanding the crosstalk mechanisms in
these signaling molecules is important but challenging. The development
of donor compounds of ROS/RNS/RSS will aid the advances in this field.
While many donors that can release one ROS/RNS/RSS have been developed,
dual donors that can release two signaling species and facilitate
their crosstalk studies are still very rare. Those limited examples
lack the ability to precisely control the timing of two releases.
In this work, a 2-methoxy-6-naphthacyl-derived tertiary SNO compound,
Naph-SNO, was designed and evaluated as the dual donor for NO and
H2S2. The 2-methoxy-6-naphthacyl structure was
demonstrated to be a novel photoremovable protecting group that could
directly uncage C–S bonds. Under the irradiation of lights
with different wavelengths (visible or UV), Naph-SNO could release
NO and H2S2 in a stepwise manner, or simultaneously
(i.e., likely producing the crosstalk product HSNO/HSSNO). In addition,
the release of payloads from the donor also produced an end product
with blue fluorescence. Therefore, the release process could be easily
monitored in “real time.” This controllable photo-triggered
release strategy has the potential to be used in the design of other
RNS/RSS dual donors.
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