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Plasmodium Cost of Resistance and Life Stage Development within the Mosquito Vector

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Hundreds of thousands of people die annually from malaria; a protozoan of the genus Plasmodium is responsible for this mortality. The Plasmodium parasite undergoes several life stages within the mosquito vector, the transition between which require passage across the lumen of the mosquito midgut. It has been observed that in…

Hundreds of thousands of people die annually from malaria; a protozoan of the genus Plasmodium is responsible for this mortality. The Plasmodium parasite undergoes several life stages within the mosquito vector, the transition between which require passage across the lumen of the mosquito midgut. It has been observed that in about 15% of parasites that develop ookinetes in the mosquito abdomen, sporozoites never develop in the salivary glands, indicating that passage across the midgut lumen is a significant barrier in parasite development (Gamage-Mendis et al., 1993). We aim to investigate a possible correlation between passage through the midgut lumen and drug-resistance trends in Plasmodium falciparum parasites. This study contains a total of 1024 Anopheles mosquitoes: 187 Anopheles gambiae and 837 Anopheles funestus samples collected in high malaria transmission areas of Mozambique between March and June of 2016. Sanger sequencing will be used to determine the prevalence of known resistance alleles for anti-malarial drugs: chloroquine resistance transporter (pfcrt), multidrug resistance (pfmdr1) gene, dihydropteroate synthase (pfdhps) and dihydrofolate reductase (pfdhfr). We compare prevalence of resistance between abdomen and head/thorax in order to determine whether drug resistant parasites are disproportionately hindered during their passage through the midgut lumen. A statistically significant difference between resistance alleles in the two studied body sections supports the efficacy of new anti-malarial gene surveillance strategies in areas of high malaria transmission.
ContributorsPhillips, Keeley Isabella (Author) / Huijben, Silvie (Thesis director) / Gile, Gillian (Committee member) / Young, Steven (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Diversity and Phylogeny of Trichonympha, a Parabasalian Symbiont of Termites

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Trichonympha is a Parabasalian symbiont of lower termites. They are characterized by their spindle-shape, length ranging from 75-150 micrometers, longitudinal flagella, as well as their true rostral tube composed of two plates of parabasal fibers. Ealy analysis through microscopy has revealed that the same species of Trichonympha could be found…

Trichonympha is a Parabasalian symbiont of lower termites. They are characterized by their spindle-shape, length ranging from 75-150 micrometers, longitudinal flagella, as well as their true rostral tube composed of two plates of parabasal fibers. Ealy analysis through microscopy has revealed that the same species of Trichonympha could be found across multiple species of termites. However, with recent phylogenetic analysis of the 18S region, it has been found that the species is actually genetically distinct and that Trichonympha have coevolved with their termite hosts. In this study, Trichonympha cells from Reticulitermes tibialis, R. flavipes, and R. lucifugus were isolated. DNA from the single cell samples were purified and amplified through a series of polymerase chain reactions (PCR) and gel electrophoresis. The amplified DNA was extracted from the gel and cloning was performed using competent E. coli. The colonies that formed from each sample were collected and those that amplified after a PCR reaction were sent to the ASU Genomics Core for sanger sequencing. The sequences were assembled, cleaned, and aligned and the maximum likelihood and Bayesian phylogenetic trees were constructed. It was found that there is evidence of multiple species of Trichonympha in R. flavipes and that there were multiple species found in R. lucifugus. However, the Trichonympha sequences from R. lucifugus branch in different clades which indicates that they are more distant species than those of R. flavipes. Additionally, Trichonympha cells from R. tibialis branched in the same clade as those from R. flavipes which provides evidence that the same Trichonympha species can be found in multiples species of Reticulitermes. With these pieces of evidence, we can see multiple patterns of diversity of Trichonympha in their termite hosts.
ContributorsGaylor, Maya (Author) / Gile, Gillian (Thesis director) / De Martini, Francesca (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2022-05