Wastewater-based epidemiology (WBE) has become a valuable tool for monitoring SARS-CoV-2 infection trends throughout the COVID-19 pandemic. Population biomarkers that measure the relative human fecal contribution to normalize SARS-CoV-2 wastewater concentrations are needed for improved analysis and interpretation of community infection trends. The Centers for Disease Control and Prevention National Wastewater Surveillance System (CDC NWSS) recommends using the wastewater flow rate or human fecal indicators as population normalization factors. However, there is no consensus on which normalization factor performs best. In this study, we provided the first multistate assessment of the effects of flow rate and human fecal indicators (crAssphage, F+ Coliphage, and PMMoV) on the correlation of SARS-CoV-2 wastewater concentrations and COVID-19 cases using the CDC NWSS dataset of 182 communities across six U.S. states. Flow normalized SARS-CoV-2 wastewater concentrations produced the strongest correlation with COVID-19 cases. The correlation from the three human fecal indicators were significantly lower than flow rate. Additionally, using reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) significantly improved correlation values over samples that were analyzed with real-time reverse transcription quantitative polymerase chain reaction (rRT-qPCR). Our assessment shows that utilizing flow normalization with RT-ddPCR generate the strongest correlation between SARS-CoV-2 wastewater concentrations and COVID-19 cases.
OBJECTIVES/GOALS: The goal of this project is to apply wastewater-based epidemiology to determine the trends of NTS through analysis of influent wastewater, and to isolate and identify circulating serotypes among samples that are representative of the human population in Gainesville, FL. METHODS/STUDY POPULATION: This study is being conducted in Gainesville, FL, a suburban city in North Central Florida with approximately 133,997 residents served by two wastewater reclamation facilities (WRF). Fifty-three weekly influent composite wastewater samples were collected between September 2020 and September 2021 from each WRF. Wastewater samples were processed using an adsorption-based sample concentration method and invA genetic target for NTS DNA detection and quantification via qPCR. The quantified NTS DNA wastewater concentration from each WRF was summed to determine the overall concentration. Bacterial isolation was conducted on stored wastewater of qPCR positive samples using EPA method 1682. Recovered isolate serotypes were identified through whole-genome sequence analysis. RESULTS/ANTICIPATED RESULTS: Overall, NTS was positive in Gainesville wastewater in 48/53 weeks between September 2020 – September 2021. 83/106 (78%) samples tested positive for NTS from both WRFs. The mean wastewater concentration from each WRF was 3.53 (Main St.) and 3.31 Log10 invA Genetic Copies/L. The overall mean city concentration was 6.85 Log10 invA Genetic Copies/L. There were sustained periods of NTS in the wastewater during the months of December-February and May-August. There was no observed seasonal trend in NTS wastewater concentrations over the study period. Culture-based methods will provide a comprehensive review of circulating serotypes that are responsible for both clinical and subclinical infections within the community. DISCUSSION/SIGNIFICANCE: Florida has an incidence of NTS twice the national average. Clinical surveillance is the current standard for community surveillance of NTS. Only 28% of NTS cases are identified in a clinical setting, leaving a major gap in community surveillance. This project has helped fill this gap and provided a comprehensive community-level assessment of NTS.
Introduction In the setting of hypotension, serum cortisol levels are obtained to eliminate adrenal insufficiency as a contributing cause. Morbidity, prolonged length of intensive care unit and mechanical ventilator dependent days with higher mortality rates have been associated with adrenal insufficiency. The primary objective was to identify patient characteristics and treatment indications, approaches, and outcomes in adult burn – injured patients diagnosed with adrenal insufficiency. Methods This is a single center, five-year retrospective medical record review of adult patients aged 18 years and older admitted to the burn service were included. The data was analyzed using simple and correlational statistics to describe categorical data. Logistic regression, One-way ANOVA, and univariate analyses were conducted in Statistical Analysis System (SAS)® v 9.4. Results Sixty nine patients were included, 64% male, 51years old, with 27 % total body surface area, nearly half with smoke inhalation, and 32% mortality. Duration of hydrocortisone therapy in days, total dose in milligrams, duration of wean in days, and number of times therapy reinitiated were found to be (unadjusted) significantly related to intensive care length of stay, p = < 0.001. Hydrocortisone total dose in milligrams was found to be significantly related with increased vasopressor hours, p = 0.00927, p = 0.0163. Conclusions Hypotension, leading to shock, directly correlates with overall burn survivability. Functional adrenal insufficiency may result for shock and sepsis. Hypotension was the instigating factor for hydrocortisone therapy. Random cortisol levels were obtained in all patients included in the study, with an average of 14 mcg/dl which did not exclude adrenal insufficiency as a contributing cause. Applicability of Research to Practice This exploratory research will assist with formalizing a list of triggers for initiation of adrenal replacement therapy. These results are foundational in development of outcomes resulting from treatment with varying doses of hydrocortisone and lengths of therapy.
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