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Bordetella pertussis (Bp), the causative agent of “whooping cough,” is the most important vaccine-preventable disease and an NIH and CDC high priority. The incidence rates of pertussis have increased over recent years, corresponding to the switch from the whole cell pertussis vaccine (wP) priming and acellular pertussis vaccine (aP) boosting vaccination in the early 2000s, to the current aP-only vaccination. Women given the safer but less effective aP vaccines are a rapidly increasing proportion of birthing mothers, creating a complex transition that we poorly understand and that may have particular risks to the most highly sensitive population, newborns. To protect neonates during the months before they can be fully vaccinated the CDC recommends a cocooning strategy, vaccinating all their potential contacts, creating local “herd immunity” to prevent exposure. This strategy is based on the dated and inaccurate belief that aP vaccines that protect against disease also prevent infection/transmission. In fact, there is growing evidence that aP vaccinated individuals, despite significant antiBp antibody titers, can still be colonized and can transmit Bp to close contacts, including their babies. Another approach, maternal vaccination where antibodies to Bp are transferred via placenta and colostrum/milk to the newborn, although the actual protective effect of antibody transfer is unknown. In fact, the efficacy of maternally transferred antibodies in protecting against colonization or disease has not been determined clinically. Such a clinical study would be challenging and costly and has not been attempted. The experimental evidence indicates that antibodies alone do little to prevent disease. Therefore, we are effectively in the middle of a nationwide test of the differential effects of wP-primed and aP-primed vaccination of mothers on the protection conferred to their offspring without a complete understanding of the likely impact. To address this urgent knowledge gap, we recently developed a novel mouse neonatal experimental infection system that more accurately models the unique features of the human neonatal immune system, enabling us to probe vaccine-induced protection against Bp transferred from mother to offspring. Our exciting preliminary data demonstrate that both the wP and aP maternal vaccinations confer substantial protection to the offspring’s lungs. These results demonstrate that we can measure the profound effects of maternal vaccination on the protection of pups. This approach allows us the specific means to examine the mechanisms of differentially conferred protection, as proposed below. Here we will utilize a combination of innovative immunological techniques to address the central hypothesis that transferred maternal antibodies contribute to protection in neonates via mechanisms we can distinguish using tools unique to the mouse model.

Funder: National Institutes of Health 

Amount: $393,345 

PI: Eric Harvill, College of Veterinary Medicine 

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Notable Grants

The objectives for this project will be: 1) measuring phenological shifts of these key pests and evaluating reliability of available monitoring tools and models; 2) refining IPM tools to ensure compatibility with evolving production practices, regulatory changes, and climate; 3) suppress pest populations across orchard agroecosystems with promising biocontrol agents and integrating enhanced IPM tactics; and 4) assessing the socioeconomic impact of enhanced IPM programs to promote grower adoption and market flexibility. Approach: The insect pests identified as of highest concern by Eastern growers include codling moth (Cydia pomonella), oriental fruit moth (Grapholita molesta), San Jose scale (Comstockaspis perniciosus), plum curculio (Conotrachelus nenuphar), invasive brown marmorated stink bug (Halyomorpha halys), and trunk-boring sesiid moth pests: peachtree borer (Synanthedon exitiosa), lesser peachtree borer (Synanthedon picitipes), and dogwood borer (Synanthedon scitula). For objective 1, we will use historical data sets and novel automated traps to improve decision support for codling moth and oriental fruit moth. Historical data on San Jose Scale activity and horticultural stress indicators will be integrated into novel prediction tools. Novel attractants for plum curculio and relationships between brown marorated stink bug and Samurai wasp captures will be evaluated in the field. Factors associated with grower perception of IPM tools that enable or create barriers to adoption will be quantified. For objective 2, we will conduct semi-field and field studies aimed at improving biocontrol agents for Lepidopteran pests, San Jose scale and brown marmorated stink bug. Novel management tools including scalable mating disruption for lepidopteran pests and San Jose scale will be developed in a series of field studies. Border-based management tools for plum curculio and brown marmorated stink bug including the use of deterrents and trap-based treatment thresholds will be developed using field studies. Grower perceptions of novel management tools will be captured. For objective 3, we will develop a risk-based decision matrix for pests of Eastern tree fruit based on risk levels quantified using field data. Integration of enhanced IPM tactics will be conducted in the field and compared with standard practices. In objective 4, our team will produce Extension-ready case studies highlighting the economics of enhanced IPM, hold workshops for growers and emerging farmer audiences, and mentor graduate students in Extension. Climate-smart recommendations will be delivered to Eastern tree fruit growers through field days, print, and digital media.

Funder: United States Department of Agriculture Agricultural Research Service 

Amount: $439,984 

PI: Brett Blaauw, College of Agricultural and Environmental Sciences 

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Notable Grants

Chronic environmental exposure to neurotoxic levels of the metal manganese (Mn) affects the basal ganglia system and has been linked to the etiology of Parkinsonism. Within the basal ganglia (BG), Mn primarily accumulates in the globus pallidus, resulting in dysregulation of the BG neural circuitry. As the most abundant glial cells in the CNS, astrocytes are critical for healthy brain function and are highly sensitive to Mn toxicity as the metal preferentially accumulates in astrocytes. For any neurotoxicological triggers, astrocytes are early responders and undergo a process known as “reactive astrogliosis”, characterized by morphological, molecular, and functional changes. In this context, increasing evidence suggests that Mn neurotoxicity elicits proinflammatory responses in astrocytes. However, the fundamental molecular neurotoxicological mechanisms regulating the transformation of an astrocyte’s homeostatic phenotype from quiescent to pro-inflammatory and the exact molecular regulators of astrocyte crosstalk with neurons and microglia in metal neurotoxicity remain enigmatic. In the emerging field of epitranscriptomics, the role of N6-methyladenosine (m6A) RNA modification, under the dynamic control of N6-methyltransferases, demethylases, and readers (m6A mRNA-binding proteins), in neurodegenerative diseases (NDs) is currently being explored. We recently obtained exciting new data showing that the m6A reader protein YTHDF2 is specifically downregulated by Mn in cultured human astrocyte cells and primary murine astrocytes as well as in animal models. Functional loss- and gain-of-function studies reveal an anti-inflammatory role for YTHDF2 in Mn-stimulated astrocytes, leading to a novel hypothesis that YTHDF2 may be a critical regulator impacting Mn neurotoxicity. Additional mechanistic studies identified SEK1 mRNA as a direct target of YTHDF2 and also discovered the ability of YTHDF2 to suppress the SEK1/JNK/cJun pro-inflammatory signaling axis and subsequent production and secretion of selected pro-inflammatory mediators in reactive astrocytic cultures. Very recently, we also observed a significant increase in the m6A readers YTHDF1 and YTHDF3, which can boost the translation of their respective targets. Thus, the following specific aims will be pursued to further expand our novel findings: (i) elucidate in animal models if chronic Mn exposure dysregulates YTHDF proteins in astrocytes, and whether this dysregulation contributes to Mn neurotoxicity by disrupting neuroimmune homeostasis; (ii) elucidate the pro-inflammatory signaling mechanisms underpinning YTHDF dysregulation in activated astrocytes during Mn neurotoxicity in animal models; and (iii) validate YTHDF proteins as key regulators of astrocyte-mediated immune and neuronal dysfunction in Mn neurotoxicity. Overall, we anticipate that our proposed studies will provide novel mechanistic insights into reactive astrocytic activation and neuroimmune interactions in metal neurotoxicology and will offer novel therapeutic targets to dampen the neuroinflammatory processes in environmental factor-related neurodegenerative diseases.

Funder: National Institutes of Health 

Amount: $441,529 

PI: Anumantha Kanthasamy, College of Veterinary Medicine 

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Notable Grants

Neuroinflammation is a driving force contributing to neurodegenerative diseases, including Parkinson’s disease (PD). Microglia are the primary immune cell of the brain and are among the first responders to infection, toxic insult and aggregated proteins and contribute significantly to neuroinflammation and neurodegeneration. The microglial inflammatory response, including inflammasome activation, has been demonstrated to be significantly associated with PD. Paraquat (PQ) is a commonly used herbicide that has been linked to increased risk for PD. PQ-induced neurodegeneration is tightly coupled to the activation of microglia and appears to require priming of the microglial response. Therefore, factors that modulate the microglial inflammatory response could lead to neuroprotection and slow the progression of neurodegenerative diseases. However, to date, no antiinflammatory drugs have proven successful in human clinical trials necessitating research on new targets. Hv1 (HVCN1) is a voltage-gated proton channel highly expressed on microglia in the brain and in other immune cells in the body. This proton channel regulates the activity of NADPH oxidase and production of reactive oxygen species in immune cells and especially microglia. Our preliminary data demonstrate that PQ directly increases Hv1 levels in microglia, possibly through an epigenetic mechanism involving histone acetylation. Further, our data demonstrate effects of PQ on the NLRP3 inflammasome that appear to be regulated by Hv1, providing a potential mechanism contributing to PQ-induced microglial priming. This proposal seeks to test the hypothesis that Hv1 regulates priming of microglia following PQ exposure through the NLRP3 inflammasome, leading to neuroinflammation and neurodegeneration. The Specific Aims of this project are to 1) Determine mechanisms of Hv1 regulation following paraquat exposure 2) Define the role of Hv1 in regulation of the NLRP3 inflammasome following paraquat exposure and 3) Determine the contribution of microglial Hv1 and the NLRP3 inflammasome in regulating neurodegeneration following paraquat exposure. Completion of these Aims will define regulatory mechanisms for Hv1 and determine the role of Hv1 in regulation of the NLRP3 inflammasome activation and their role in PQ-induced neuroinflammation and neurotoxicity. Together, these Aims will provide crucial information on the function of a novel regulator of neuroinflammation, Hv1, and determine whether targeting Hv1 may be a viable therapeutic strategy in toxicant-induced neurodegeneration. We will use the regional scale model as boundary conditions for two high resolution mine-specific models (Mission and Amelia mines). We will use these models to estimate impacts of mining on groundwater flows, water table elevations, and the dispersal of solutes under future climate scenarios.

If available, we will review the hydrologic model(s) prepared by Twin Pines’ consultants in order to understand key assumptions, model construction, and data sets and to possibly guide enhancements of our model.

Funder: National Institutes of Health 

Amount: $625,498 

PI: Jason Richardson, College of Veterinary Medicine 

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Notable Grants

Plasmodium vivax (Pv) is responsible for a significant portion of the malaria cases outside of Africa and causes significant socioeconomic burden worldwide. New therapies to treat Pv are needed, but a continuous, high yield culture system that enables the study of Pv’s biology has not been established. Without this tool, development of new therapies is stymied. The long-term goal of this research program is to study Pv biology to inform the development of new treatments for Pv. The objectives of this proposal are to (1) establish continuous, high yield culture systems for Pv’s asexual stages and (2) to use this culture system to test triggers of gametocytogenesis so gametocytes can be generated in vitro for study. This research is needed because it will yield a key tool to study Pv asexual and sexual stages, thereby, enabling mechanistic research to identify and develop new therapies. The major bottlenecks to establishing continuous cultures have been (1) Pv preferentially invades a specific subset of reticulocytes, or immature erythrocytes, in culture, and (2) low parasite numbers due to poor development even when reticulocytes are supplemented for invasion. In preliminary studies, our team has used P. cynomolgi (Pcy), a closely-related malaria parasite to Pv, to develop a strategy for addressing these obstacles. We have shown that Pcy strains vary in their ability to grow in culture with some strains growing in culture immediately whereas others adapt with time, suggesting that intrinsic variation within strains exists and can be selected for to establish continuous cultures. Using our Pcy strains that grow in culture, we have also shown that a bone marrow (BM) mimetic composed of secreted factors from BM mesenchymal stromal/stem cells (MSCs) significantly improves Pcy development in culture. Intriguingly, our data also indicate that BM reticulocytes may trigger the development of gametocytes. Based on these data and the prior literature, we propose to test the hypothesis that continuous cultures of Pv can be achieved by establishing Pv strains that invade multiple reticulocyte subsets in culture using conditions that mimic the BM where Pv thrives in vivo. This proposal will test our hypothesis by pursuing three specific aims. During the R61 phase, the experiments in Aim 1 will establish Pv strain(s) that robustly invade reticulocytes in culture while the experiments in Aim 2 will improve Pv development in vitro by mimicking the BM microenvironment. Together, these aims are expected to yield a continuous culture system for Pv asexual stages. After achieving this milestone, Aim 3 will use this culture system to test triggers of gametocytogenesis during the R33 phase. The expected outcomes of this research are a continuous culture system for Pv and new insight into Pv invasion, development and gametocytogenesis. This research will have an immediate impact on Pv research because it will provide essential tools that will enable mechanistic insight into Pv biology and provide new knowledge that may lead to new therapies.

Funder: National Institutes of Health 

Amount: $1,152,630 

PI: Chester Joyner, College of Veterinary Medicine 

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Notable Grants

Nematodes are among the top yield-robbers encountered by soybean farmers across the U.S. Almost all SCN resistant varieties in U.S. soybean production are derived from two genetic sources, imposing risk of genetic vulnerability and resistance breakdown. Soybean breeders and growers are limited in genetic sources for nematode resistance with competitive yield, which places U.S. soybean production at risk. Discovery of novel nematode resistance from genetically diverse sources is essential for sustainable soybean production. Deployment of resistant varieties will improve U.S. soybean production and protect yields and farmers’ income.

Our team has successfully developed a strong pipeline of soybean germplasm with resistance to SCN or RKN and competitive yield across all maturity groups. Based on previous discoveries, many genetic populations have been developed to confirm and deploy these genes for nematode resistance. In this new proposal, this team will use an integrated conventional and advanced genomic technologies to achieve the following objectives: 1) incorporate nematode resistance genes into elite high-yielding lines to develop SCN and/or RKN resistant soybean varieties in MG 0 through VIII; 2) identify novel sources of multiple nematode resistance from existing resistant sources or soybean germplasm from USDA Soybean Germplasm Collections; 3) map nematode resistance gene(s) to develop DNA markers for efficient breeding selection; and 4) incorporate SCN and/or RKN resistance into high-yielding lines with value-added seed composition or other key abiotic and biotic tolerance traits.

This work will benefit the entire value chain by providing new soybean varieties adapted to local growing conditions with resistance to multiple nematode species. This project will also provide new and improved materials to commercial and public breeders for use as parental stocks to develop high-yielding, nematode resistant varieties. DNA markers and QTL information generated will benefit soybean researchers, enabling marker-assisted selection and seeking understanding of the genetic mechanisms underlying nematode resistance.

Funder: U.S. Department of Energy

Amount: $333,489

PI: Zenglu Li, College of Agricultural and Environmental Sciences

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Notable Grants

Even in fungal model systems, up to 40% of genes have unknown or poorly characterized functions with no informative homologies in databases. Fungi tolerate a variety of stressful environments, including a wide range of temperature and moisture levels. The same fungal species isolated from different environments often shows variation in its genetic complement. Studies of pan-genomes (the full genetic complement of a species assembled from varied individuals) have identified core genes (present in all isolates), accessory genes (present in >1 isolate) and singleton genes (present in only 1 isolate). Accessory and singleton genes are much more likely to have an unknown or poorly characterized function, presumably because these genes have evolved or been retained only in a subset of environments.

We propose to exploit natural variation in two fungal species to identify genes of unknown function that allow them to survive across temperature and moisture ranges. We have previously collected isolates of the filamentous fungus Aspergillus fumigatus and the yeast Saccharomyces paradoxus from a range of urban, natural, and agricultural environments in North America. Our collection sites spanned a wide range of temperature and moisture levels, and we already have WGS data for many of the isolates.

We propose to construct pan-genomes for A. fumigatus and S. paradoxus and identify core, accessory, and singleton genes and genes of unknown or poorly characterized function. We will pick environmental isolates of A. fumigatus and S. paradoxus representing the environments with the most extreme temperatures (coldest and warmest) and humidity (driest and wettest) that also have the most genes of unknown or poorly characterized function. Transcriptomes will be analyzed from these strains grown under a variety of temperature and humidity levels. We will identify genes that are differentially expressed across conditions, especially those with unknown or poorly defined functions. We will create gene deletions and tags and use them in functional validation experiments for genes of unknown or poorly characterized function whose expression suggests roles in response to temperature and/or humidity variation.

Our ultimate goals are to reduce the number of genes of unknown/poorly characterized function in the filamentous fungus A. fumigatus and the yeast S. paradoxus and to discover new genes that are important for tolerance to a range of temperature and water-availability levels in these fungi. If this approach proves successful, it can be used for gene discovery in other fungi, including nonmodel systems that lack genetic tools and will allow improved prediction of the resilience of other species of environmental fungi. Though it is currently appreciated that there is environmental variation in species, this variation has not been used in the way we propose to assign gene function. Thus, the proposed experiments will serve as a guide for future work leveraging natural variation and phenotyping to assign function to fungal genes of unknown function, especially those that allow fungi to thrive in hostile environments. Understanding such genes will be helpful to the Army because fungi are notorious for degrading materials in the challenging environments where the Army’s missions often occur.

Funder: U.S. Department of the Army

Amount: $449,999

PI: Michelle Momany, Franklin College of Arts and Sciences, Department of Plant Biology

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Notable Grants

We propose to continue development of a groundwater-hydrologic model for southeast Georgia and surrounding region (Okefenokee Swamp and associated rivers). We will explore the role of groundwater flows across southeast Georgia for future climate and coastal development scenarios including the effective dispersion of scalars within the groundwater system. We will maintain a specific focus on net flow from groundwater into/out of the Okefenokee Swamp, Trail Ridge, and the St. Marys River and how this net flow is expected to change under future conditions. The model will allow us to quantify the fate of groundwater and solutes in the system. With the models we will be able to estimate impacts of mining on water levels and dispersion of solutes (tailings) in both groundwater and surface waters with particular focus on solutes potentially entering the Floridian aquifer, the Okefenokee Swamp, and the St. Marys River; how Floridan Aquifer withdrawals may influence the Okefenokee Swamp; and how mine operations and reclamation may affect the characteristics of the Surficial Aquifer and how such changes may influence the hydrologic characteristics of Trail Ridge and the Okefenokee Swamp.. Our initial

focus will be on the surficial and Floridian aquifer and their interaction with the Okefenokee Swamp and associated rivers. We will use the numerical model outputs to develop conceptual models of the water cycle in the region and estimates of solute dispersion and changes in surface water levels.

We will use the regional scale model as boundary conditions for two high resolution mine-specific models (Mission and Amelia mines). We will use these models to estimate impacts of mining on groundwater flows, water table elevations, and the dispersal of solutes under future climate scenarios.

If available, we will review the hydrologic model(s) prepared by Twin Pines’ consultants in order to understand key assumptions, model construction, and data sets and to possibly guide enhancements of our model.

Funder: Chemours Company

Amount: $552,950

PI: C. Brock Woodson, College of Engineering

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The overall objective of this project is to improve the management of whiteflies and whitefly-transmitted viruses in vegetable crops in Georgia and other southeastern states. The whitefly, Bemisia tabaci, causes global economic losses including an epidemic by this pest and whitefly-transmitted viruses that severely impact vegetable and other crop production in the southeastern United States. The rapid evolution of insecticide-resistance in whitefly populations makes it unsustainable to use chemical control. We will investigate whiteflies and whitefly-transmitted viruses in vegetable cropping systems from the perspective of an ecology-based integrated pest-plant-virus management system.

Funder: USDA ARS

Amount: $4,011,522

PI: Allen Moore, College of Agricultural and Environmental Sciences

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Several methods have been developed to inactivate virus particles. These inactivation methods must be effective and reliable to prevent the accidental spread of viruses. It is crucial to completely inactivate high-risk agents, such as SARS-CoV-2, before they are removed from high-level biocontainment facilities for further handling. However, it is unclear whether common methods of virus inactivation are effective in inactivating positive-sense RNA viruses. Positive-sense RNA viruses contain RNA genomes of positive polarity and include viruses which can cause serious illness in humans and often result in epidemics and pandemics. The genome of a positive-sense RNA directly serves as messenger RNA (mRNA), thus, can be immediately translated by host cells to produce infectious virions. Therefore, if viral RNA is not properly inactivated during inactivation procedures, the intact RNA can potentially cause infection if introduced into permissive cells. This underscores the need for inactivation protocols that ensure complete inactivation of viral RNA to prevent the accidental release of virus particles into the environment.

Validation for the absence of RNA infectivity after virus inactivation is rarely performed, raising significant biosecurity risks associated with positive-sense RNA viruses. Despite the widespread use of inactivating agents in research labs, there are no standardized protocols for validating RNA infectivity. Testing methods for RNA infectivity are cumbersome and require expertise in RNA, cell culture, and transfection techniques. This study will systematically evaluate the efficacy of existing viral RNA inactivation methods, focusing on their ability to render viral RNA non-infectious. We will develop reagents and protocols required for infectivity testing of viral RNA. By employing appropriate controls and rigorous validation techniques, we will establish standardized protocols for the inactivation of positive-sense viruses and their RNA.

Funder: Centers for Disease Control and Prevention 

Amount: $846,421 

PI: Lok Joshi, College of Veterinary Medicine