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The objective of this study is to gain a comparative understanding of spatial determinants for outreach and clinic vaccination, which is critical for operationalizing efforts and breaking down structural biases; particularly relevant in countries where resources are low, and sub-region variance is high. Leveraging a massive effort to digitize public system reporting by Lady and Community Health Workers (CHWs) with geo-located data on over 4 million public-sector vaccinations from September 2017 through 2019, understanding health service operations in relation to vulnerable spatial determinants were made feasible. Location and type of vaccinations (clinic or outreach) were compared to regional spatial attributes where they were performed. Important spatial attributes were assessed using three modeling approaches (ridge regression, gradient boosting, and a generalized additive model). Consistent predictors for outreach, clinic, and proportion of third dose pentavalent vaccinations by region were identified. Of all Penta-3 vaccination records, 86.3% were performed by outreach efforts. At the tehsil level (fourth-order administrative unit), controlling for child population, population density, proportion of population in urban areas, distance to cities, average maternal education, and other relevant factors, increased poverty was significantly associated with more in-clinic vaccinations (β = 0.077), and lower proportion of outreach vaccinations by region (β = -0.083). Analyses at the union council level (fifth-administrative unit) showed consistent results for the differential importance of poverty for outreach versus clinic vaccination. Relevant predictors for each type of vaccination (outreach vs. in-clinic) show how design of outreach vaccination can effectively augment vaccination efforts beyond healthcare services through clinics. As Pakistan is third among countries with the most unvaccinated and under-vaccinated children, understanding barriers and factors associated with vaccination can be demonstrative for other national and sub-national regions facing challenges and also inform guidelines on supporting CHWs in health systems. 

Abstract The Antarctic midge, Belgica antarctica , is a wingless, non-biting midge endemic to Antarctica. Larval development requires at least 2 years, but adults live only 2 weeks. The nonfeeding adults mate in swarms and females die shortly after oviposition. Eggs are suspended in a gel of unknown composition that is expressed from the female accessory gland. This project characterizes molecular mechanisms underlying reproduction in this midge by examining differential gene expression in whole males, females, and larvae, as well as in male and female accessory glands. Functional studies were used to assess the role of the gel encasing the eggs, as well as the impact of stress on reproductive biology. RNA-seq analyses revealed sex- and development-specific gene sets along with those associated with the accessory glands. Proteomic analyses were used to define the composition of the egg-containing gel, which is generated during multiple developmental stages and derived from both the accessory gland and other female organs. Functional studies indicate the gel provides a larval food source as well as a buffer for thermal and dehydration stress. All of these function are critical to juvenile survival. Larval dehydration stress directly reduces production of storage proteins and key accessory gland components, a feature that impacts adult reproductive success. Modeling reveals that bouts of dehydration may have a significant impact on population growth. This work lays a foundation for further examination of reproduction in midges and provides new information related to general reproduction in dipterans. A key aspect of this work is that reproduction and stress dynamics, currently understudied in polar organisms, are likely to prove critical in determining how climate change will alter their survivability. 

Abstract BackgroundThe stable fly,Stomoxys calcitrans, is a major blood-feeding pest of livestock that has near worldwide distribution, causing an annual cost of over $2 billion for control and product loss in the USA alone. Control of these flies has been limited to increased sanitary management practices and insecticide application for suppressing larval stages. Few genetic and molecular resources are available to help in developing novel methods for controlling stable flies. ResultsThis study examines stable fly biology by utilizing a combination of high-quality genome sequencing and RNA-Seq analyses targeting multiple developmental stages and tissues. In conjunction, 1600 genes were manually curated to characterize genetic features related to stable fly reproduction, vector host interactions, host-microbe dynamics, and putative targets for control. Most notable was characterization of genes associated with reproduction and identification of expanded gene families with functional associations to vision, chemosensation, immunity, and metabolic detoxification pathways. ConclusionsThe combined sequencing, assembly, and curation of the male stable fly genome followed by RNA-Seq and downstream analyses provide insights necessary to understand the biology of this important pest. These resources and new data will provide the groundwork for expanding the tools available to control stable fly infestations. The close relationship ofStomoxysto other blood-feeding (horn flies andGlossina) and non-blood-feeding flies (house flies, medflies,Drosophila) will facilitate understanding of the evolutionary processes associated with development of blood feeding among the Cyclorrhapha.