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.

The symbiosis between termites and their parabasalid hindgut protists centers around the wood digestion that is needed for both species to acquire the nutrients from wood. One of the important carbohydrate-active proteins required for the wood breakdown are glycoside hydrolase (GH) families. Previous studies have looked at the phylogeny of some of these protein families from a termite whole gut transcriptome or in a different context than lignocellulose digestion. In this study, we attempt to understand the function and evolution of these GH families in the context of protist evolution by using protist single cell transcriptomes. 14 families of interest were chosen to create phylogenetic trees: GH2, GH3, GH5, GH7, GH8, GH9, GH10, GH11, GH26, GH43, GH45, GH55, GH67, GH95 for their interesting expressions across different protists such as being present in all protists or being present in only termite-associated protists. The dbCAN2 (automated Carbohydrate-active enzyme ANnotation) program was used to find GH families in each of the protist single cell transcriptomes and additional characterized sequences registered on the National Center for Biotechnology Information to create phylogenetic trees for each of the GH families of interest. Results show that many of the GH families expressed in protists were acquired through horizontal gene transfer from fungi and bacteria. Additionally, comparison to the parabasalid phylogeny indicates most GH families evolved independently from the protists. Based on the pattern of expression of these GH families throughout different protist orders, conclusions can be made about whether the specific family was vertically or horizontally acquired in the termite symbionts.

Cells have mechanisms in place to maintain the specific lipid composition of distinct organelles including vesicular transport by the endomembrane system and non-vesicular lipid transport by lipid transport proteins. Oxysterol Binding Proteins (OSBPs) are a family of lipid transport proteins that transfer lipids at various membrane contact sites (MCSs). OSBPs have been extensively investigated in human and yeast cells where twelve have been identified in Homo sapiens and seven in Saccharomyces cerevisiae. The evolutionary relationship between these well-characterized OSBPs is still unclear. Reconstructed OSBP phylogenies revealed that the ancestral Saccharomycotinan had four OSBPs, the ancestral Holomycotan had five OSBPs, the ancestral Holozoan had six OSBPs, the ancestral Opisthokont had three OSBPs, and the ancestral Eukaroyte had three OSBPs. Our analysis identified three clades of ancient OSBPs not present in animals or fungi.