Longitudinal Analysis of Urban Stormwater Microbiome and Resistome from Watersheds with and without Green Infrastructure using Long-Read Sequencing.

Since stormwater conveys a variety of contaminants into water bodies, green infrastructure (GI) is increasingly being adopted as an on-site treatment solution in addition to controlling peak flows. The purpose of this study was to identify differences in microbial water quality of stormwater in watersheds retrofitted with GI vs. those without GI. Considering stormwater is recently recognized as a contributor to the antibiotic resistance (AR) threat, another goal of this study was to characterize changes in the microbiome and collection of AR genes (resistome) of urban stormwater with season, rainfall characteristics, and fecal contamination. MinION long-read sequencing was used to analyze stormwater microbiome and resistome from watersheds with and without GI in Columbus, Ohio, United States, over 18 months. We characterized fecal contamination in stormwater via culturing Escherichia coli and with molecular microbial source tracking (MST) to identify sources of fecal contamination. Overall, season and storm event (rainfall) characteristics had the strongest relationships with changes in the stormwater microbiome and resistome. We found no significant differences in microbial water quality or the microbiome of stormwater in watersheds with and without GI implemented. However, there were differences between the communities of microorganisms hosting antibiotic resistance genes (ARGs) in stormwater from watersheds with and without GI, indicating the potential sensitivity of AR bacteria to treatment. Stormwater was contaminated with high concentrations of human-associated fecal bacterial genes, and the ARG host bacterial community had considerable similarities to human feces/wastewater. We also identified 15 potential pathogens hosting ARGs in these stormwater resistome, including vancomycin-resistant Enterococcus faecium (VRE) and multidrug-resistant Pseudomonas aeruginosa. In summary, urban stormwater is highly contaminated and has a great potential to spread AR and microbial hazards to nearby environments. This study presents the most comprehensive analysis of stormwater microbiome and resistome to date, which is crucial to understanding the potential microbial risk from this matrix. This information can be used to guide future public health policy, stormwater reuse programs, and urban runoff treatment initiatives.

Dynamic SARS-CoV-2 surveillance model combining seroprevalence and wastewater concentrations for post-vaccine disease burden estimates.

BACKGROUND: Despite wide scale assessments, it remains unclear how large-scale severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination affected the wastewater concentration of the virus or the overall disease burden as measured by hospitalization rates. METHODS: We used weekly SARS-CoV-2 wastewater concentration with a stratified random sampling of seroprevalence, and linked vaccination and hospitalization data, from April 2021-August 2021 in Jefferson County, Kentucky (USA). Our susceptible ( S ), vaccinated ( V ), variant-specific infected ( I 1 and I 2 ), recovered ( R ), and seropositive ( T ) model ( S V I 2 R T ) tracked prevalence longitudinally. This was related to wastewater concentration. RESULTS: Here we show the 64% county vaccination rate translate into about a 61% decrease in SARS-CoV-2 incidence. The estimated effect of SARS-CoV-2 Delta variant emergence is a 24-fold increase of infection counts, which correspond to an over 9-fold increase in wastewater concentration. Hospitalization burden and wastewater concentration have the strongest correlation (r = 0.95) at 1 week lag. CONCLUSIONS: Our study underscores the importance of continuing environmental surveillance post-vaccine and provides a proof-of-concept for environmental epidemiology monitoring of infectious disease for future pandemic preparedness.

It is unclear how large-scale COVID-19 vaccination impacts wastewater concentration or overall disease burden. Here, we developed a mathematical surveillance model that allows estimation of overall vaccine impact based on the amount of SARS-CoV-2 in wastewater, seroprevalence and the number of cases admitted to hospitals between April 2021­August 2021 in Jefferson County, Kentucky USA. We found that a 64% vaccination coverage correlated to a 61% decrease in COVID-19 cases. The emergence of the SARS-CoV-2 Delta variant during the time of the surveillance directly correlated with a sharp increase in infection incidence as well as viral counts in wastewater. The hospitalization burden was closely reflected by the viral count found in the wastewater, indicating that post-vaccine environmental surveillance can be an effective method of estimating changing disease prevalence in future pandemics.

Inferring delays in partially observed gene regulation processes.

MOTIVATION: Cell function is regulated by gene regulatory networks (GRNs) defined by protein-mediated interaction between constituent genes. Despite advances in experimental techniques, we can still measure only a fraction of the processes that govern GRN dynamics. To infer the properties of GRNs using partial observation, unobserved sequential processes can be replaced with distributed time delays, yielding non-Markovian models. Inference methods based on the resulting model suffer from the curse of dimensionality. RESULTS: We develop a simulation-based Bayesian MCMC method employing an approximate likelihood for the efficient and accurate inference of GRN parameters when only some of their products are observed. We illustrate our approach using a two-step activation model: an activation signal leads to the accumulation of an unobserved regulatory protein, which triggers the expression of observed fluorescent proteins. With prior information about observed fluorescent protein synthesis, our method successfully infers the dynamics of the unobserved regulatory protein. We can estimate the delay and kinetic parameters characterizing target regulation including transcription, translation, and target searching of an unobserved protein from experimental measurements of the products of its target gene. Our method is scalable and can be used to analyze non-Markovian models with hidden components. AVAILABILITY AND IMPLEMENTATION: Our code is implemented in R and is freely available with a simple example data at https://github.com/Mathbiomed/SimMCMC.

Household environment and animal fecal contamination are critical modifiers of the gut microbiome and resistome in young children from rural Nicaragua.

BACKGROUND: Early life plays a vital role in the development of the gut microbiome and subsequent health. While many factors that shape the gut microbiome have been described, including delivery mode, breastfeeding, and antibiotic use, the role of household environments is still unclear. Furthermore, the development of the gut antimicrobial resistome and its role in health and disease is not well characterized, particularly in settings with water insecurity and less sanitation infrastructure. RESULTS: This study investigated the gut microbiome and resistome of infants and young children (ages 4 days-6 years) in rural Nicaragua using Oxford Nanopore Technology's MinION long-read sequencing. Differences in gut microbiome diversity and antibiotic resistance gene (ARG) abundance were examined for associations with host factors (age, sex, height for age z-score, weight for height z-score, delivery mode, breastfeeding habits) and household environmental factors (animals inside the home, coliforms in drinking water, enteric pathogens in household floors, fecal microbial source tracking markers in household floors). We identified anticipated associations of higher gut microbiome diversity with participant age and vaginal delivery. However, novel to this study were the significant, positive associations between ruminant and dog fecal contamination of household floors and gut microbiome diversity. We also identified greater abundance of potential pathogens in the gut microbiomes of participants with higher fecal contamination on their household floors. Path analysis revealed that water quality and household floor contamination independently and significantly influenced gut microbiome diversity when controlling for age. These gut microbiome contained diverse resistome, dominated by multidrug, tetracycline, macrolide/lincosamide/streptogramin, and beta-lactam resistance. We found that the abundance of ARGs in the gut decreased with age. The bacterial hosts of ARGs were mainly from the family Enterobacteriaceae, particularly Escherichia coli. CONCLUSIONS: This study identified the role of household environmental contamination in the developing gut microbiome and resistome of young children and infants with a One Health perspective. We found significant relationships between host age, gut microbiome diversity, and the resistome. Understanding the impact of the household environment on the development of the resistome and microbiome in early life is essential to optimize the relationship between environmental exposure and human health. Video Abstract.

Immune-Modulating Lipid Nanomaterials for the Delivery of Biopharmaceuticals.

In recent years, with the approval of preventative vaccines for pandemics, lipid nanoparticles have become a prominent RNA delivery vehicle. The lack of long-lasting effects of non-viral vectors is an advantage for infectious disease vaccines. With the introduction of microfluidic processes that facilitate the encapsulation of nucleic acid cargo, lipid nanoparticles are being studied as delivery vehicles for various RNA-based biopharmaceuticals. In particular, using microfluidic chip-based fabrication processes, nucleic acids such as RNA and proteins can be effectively incorporated into lipid nanoparticles and utilized as delivery vehicles for various biopharmaceuticals. Due to the successful development of mRNA therapies, lipid nanoparticles have emerged as a promising approach for the delivery of biopharmaceuticals. Biopharmaceuticals of various types (DNA, mRNA, short RNA, proteins) possess expression mechanisms that are suitable for manufacturing personalized cancer vaccines, while also requiring formulation with lipid nanoparticles. In this review, we describe the basic design of lipid nanoparticles, the types of biopharmaceuticals used as carriers, and the microfluidic processes involved. We then present research cases focusing on lipid-nanoparticle-based immune modulation and discuss the current status of commercially available lipid nanoparticles, as well as future prospects for the development of lipid nanoparticles for immune regulation purposes.

Wastewater and seroprevalence for pandemic preparedness: variant analysis, vaccination effect, and hospitalization forecasting for SARS-CoV-2, in Jefferson County, Kentucky.

Despite wide scale assessments, it remains unclear how large-scale SARS-CoV-2 vaccination affected the wastewater concentration of the virus or the overall disease burden as measured by hospitalization rates. We used weekly SARS-CoV-2 wastewater concentration with a stratified random sampling of seroprevalence, and linked vaccination and hospitalization data, from April 2021-August 2021 in Jefferson County, Kentucky (USA). Our susceptible (S), vaccinated (V), variant-specific infected I1 and I2, recovered (R), and seropositive (T) model SVI2RT tracked prevalence longitudinally. This was related to wastewater concentration. The 64% county vaccination rate translated into about 61% decrease in SARS-CoV-2 incidence. The estimated effect of SARS-CoV-2 Delta variant emergence was a 24-fold increase of infection counts, which corresponded to an over 9-fold increase in wastewater concentration. Hospitalization burden and wastewater concentration had the strongest correlation (r = 0.95) at 1 week lag. Our study underscores the importance of continued environmental surveillance post-vaccine and provides a proof-of-concept for environmental epidemiology monitoring of infectious disease for future pandemic preparedness.

Relationship between freshwater harmful algal blooms and neurodegenerative disease incidence rates in South Korea.

BACKGROUND: Due to anthropogenic activities and global warming, the severity and distribution of harmful algal blooms (HABs) have been increasing steadily worldwide, including in South Korea (S. Korea). Previous studies reported that exposure to HABs could increase the risk of HAB-related diseases. However, very few studies examined the linkage between HABs and disease occurrence, particularly in S. Korea. The objective of this study was to evaluate the potential impact of HABs on neurodegenerative diseases (NDs), including Alzheimer's disease, Parkinson's disease, and motor neuron disease, at a population level. METHODS: Thirteen-year data (2005-2017) for chlorophyll-a (chl-a) concentrations as a bloom-related parameter, annual numbers of NDs, and population information were collected. First, the entire area of S. Korea was divided into a grid of 1 km, and the population number in each 1-km grid was collected using the Statistical Geographic Information Service Plus system. Cross-sectional time series data were analyzed with two statistical models, a generalized linear mixed model and a generalized linear model. RESULTS: The results show a general trend of increasing chl-a concentration and NDs year by year. We observed positive correlations between HAB intensity and the incidence rate of NDs. Particularly, HABs seem to have the most long-term carry-over effect on Parkinson's disease. Another key finding was that a 5-km radius from the HAB location was the boundary that showed the most significant associations with three NDs. CONCLUSIONS: This study provides statistical evidence that supports the potential risk of NDs from the exposure to HAB. Thus, it is recommended to monitor a broad spectrum of cyanotoxins, including neurotoxins, in bloom-affected regions in S. Korea and epidemiological studies in the future.

Quantifying the relationship between sub-population wastewater samples and community-wide SARS-CoV-2 seroprevalence.

Robust epidemiological models relating wastewater to community disease prevalence are lacking. Assessments of SARS-CoV-2 infection rates have relied primarily on convenience sampling, which does not provide reliable estimates of community disease prevalence due to inherent biases. This study conducted serial stratified randomized samplings to estimate the prevalence of SARS-CoV-2 antibodies in 3717 participants, and obtained weekly samples of community wastewater for SARS-CoV-2 concentrations in Jefferson County, KY (USA) from August 2020 to February 2021. Using an expanded Susceptible-Infected-Recovered model, the longitudinal estimates of the disease prevalence were obtained and compared with the wastewater concentrations using regression analysis. The model analysis revealed significant temporal differences in epidemic peaks. The results showed that in some areas, the average incidence rate, based on serological sampling, was 50 % higher than the health department rate, which was based on convenience sampling. The model-estimated average prevalence rates correlated well with the wastewater (correlation = 0.63, CI (0.31,0.83)). In the regression analysis, a one copy per ml-unit increase in weekly average wastewater concentration of SARS-CoV-2 corresponded to an average increase of 1-1.3 cases of SARS-CoV-2 infection per 100,000 residents. The analysis indicates that wastewater may provide robust estimates of community spread of infection, in line with the modeled prevalence estimates obtained from stratified randomized sampling, and is therefore superior to publicly available health data.

Modeling Incorporating the Severity-Reducing Long-term Immunity: Higher Viral Transmission Paradoxically Reduces Severe COVID-19 During Endemic Transition.

Natural infection with severe acute respiratory syndrome-coronavirus-2 or vaccination induces virus-specific immunity protecting hosts from infection and severe disease. While the infection-preventing immunity gradually declines, the severity-reducing immunity is relatively well preserved. Here, based on the different longevity of these distinct immunities, we develop a mathematical model to estimate courses of endemic transition of coronavirus disease 2019 (COVID-19). Our analysis demonstrates that high viral transmission unexpectedly reduces the rates of progression to severe COVID-19 during the course of endemic transition despite increased numbers of infection cases. Our study also shows that high viral transmission amongst populations with high vaccination coverages paradoxically accelerates the endemic transition of COVID-19 with reduced numbers of severe cases. These results provide critical insights for driving public health policies in the era of 'living with COVID-19.'

Beyond the Michaelis-Menten: Bayesian Inference for Enzyme Kinetic Analysis.

Although the Michaelis-Menten (MM) rate law has been widely used to estimate enzyme kinetic parameters, it works only under the condition of extremely low enzyme concentration. Furthermore, even when this condition is satisfied, parameter estimation is often imprecise due to the parameter identifiability issue. To overcome these limitations of the canonical approach to enzyme kinetics, we developed a Bayesian approach based on a modified form of the MM rate law, which is derived with the total quasi-steady state approximation. Here, we illustrate how to perform the Bayesian inference for the progress curve assay with our user-friendly computational R package. We also describe an optimal experimental design for the progress curve assay, with which enzyme kinetic parameters can be accurately and precisely estimated from minimal measurements of the progress curves.

An interaction Neyman-Scott point process model for coronavirus disease-19.

With rapid transmission, the coronavirus disease 2019 (COVID-19) has led to over three million deaths worldwide, posing significant societal challenges. Understanding the spatial patterns of patient visits and detecting local cluster centers are crucial to controlling disease outbreaks. We analyze COVID-19 contact tracing data collected from Seoul, which provide a unique opportunity to understand the mechanism of patient visit occurrence. Analyzing contact tracing data is challenging because patient visits show strong clustering patterns, while cluster centers may have complex interaction behavior. Cluster centers attract each other at mid-range distances because other cluster centers are likely to appear in nearby regions. At the same time, they repel each other at too small distances to avoid merging. To account for such behaviors, we develop a novel interaction Neyman-Scott process that regards the observed patient visit events as offsprings generated from a parent cluster center. Inference for such models is challenging since the likelihood involves intractable normalizing functions. To address this issue, we embed an auxiliary variable algorithm into our Markov chain Monte Carlo. We fit our model to several simulated and real data examples under different outbreak scenarios and show that our method can describe the spatial patterns of patient visits well. We also provide useful visualizations that can inform public health interventions for infectious diseases, such as social distancing.

Hierarchical Bayesian models of transcriptional and translational regulation processes with delays.

MOTIVATION: Simultaneous recordings of gene network dynamics across large populations have revealed that cell characteristics vary considerably even in clonal lines. Inferring the variability of parameters that determine gene dynamics is key to understanding cellular behavior. However, this is complicated by the fact that the outcomes and effects of many reactions are not observable directly. Unobserved reactions can be replaced with time delays to reduce model dimensionality and simplify inference. However, the resulting models are non-Markovian, and require the development of new inference techniques. RESULTS: We propose a non-Markovian, hierarchical Bayesian inference framework for quantifying the variability of cellular processes within and across cells in a population. We illustrate our approach using a delayed birth-death process. In general, a distributed delay model, rather than a popular fixed delay model, is needed for inference, even if only mean reaction delays are of interest. Using in silico and experimental data we show that the proposed hierarchical framework is robust and leads to improved estimates compared to its non-hierarchical counterpart. We apply our method to data obtained using time-lapse microscopy and infer the parameters that describe the dynamics of protein production at the single cell and population level. The mean delays in protein production are larger than previously reported, have a coefficient of variation of around 0.2 across the population, and are not strongly correlated with protein production or growth rates. AVAILABILITY AND IMPLEMENTATION: Accompanying code in Python is available at https://github.com/mvcortez/Bayesian-Inference. CONTACT: kresimir.josic@gmail.com or jaekkim@kaist.ac.kr or cbskust@korea.ac.kr. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Survival dynamical systems: individual-level survival analysis from population-level epidemic models.

In this paper, we show that solutions to ordinary differential equations describing the large-population limits of Markovian stochastic epidemic models can be interpreted as survival or cumulative hazard functions when analysing data on individuals sampled from the population. We refer to the individual-level survival and hazard functions derived from population-level equations as a survival dynamical system (SDS). To illustrate how population-level dynamics imply probability laws for individual-level infection and recovery times that can be used for statistical inference, we show numerical examples based on synthetic data. In these examples, we show that an SDS analysis compares favourably with a complete-data maximum-likelihood analysis. Finally, we use the SDS approach to analyse data from a 2009 influenza A(H1N1) outbreak at Washington State University.

Bayesian inference of distributed time delay in transcriptional and translational regulation.

MOTIVATION: Advances in experimental and imaging techniques have allowed for unprecedented insights into the dynamical processes within individual cells. However, many facets of intracellular dynamics remain hidden, or can be measured only indirectly. This makes it challenging to reconstruct the regulatory networks that govern the biochemical processes underlying various cell functions. Current estimation techniques for inferring reaction rates frequently rely on marginalization over unobserved processes and states. Even in simple systems this approach can be computationally challenging, and can lead to large uncertainties and lack of robustness in parameter estimates. Therefore we will require alternative approaches to efficiently uncover the interactions in complex biochemical networks. RESULTS: We propose a Bayesian inference framework based on replacing uninteresting or unobserved reactions with time delays. Although the resulting models are non-Markovian, recent results on stochastic systems with random delays allow us to rigorously obtain expressions for the likelihoods of model parameters. In turn, this allows us to extend MCMC methods to efficiently estimate reaction rates, and delay distribution parameters, from single-cell assays. We illustrate the advantages, and potential pitfalls, of the approach using a birth-death model with both synthetic and experimental data, and show that we can robustly infer model parameters using a relatively small number of measurements. We demonstrate how to do so even when only the relative molecule count within the cell is measured, as in the case of fluorescence microscopy. AVAILABILITY AND IMPLEMENTATION: Accompanying code in R is available at https://github.com/cbskust/DDE_BD. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Harmful algal blooms and liver diseases: focusing on the areas near the four major rivers in South Korea.

Freshwater harmful algal blooms (HABs) have become a global concern because blooms contain cyanotoxins that can cause liver damage and other negative health impacts. In South Korea, HABs have been frequently observed along the major rivers (Han, Geum, Nakdong, and Youngsan) in recent years. However, there are hardly any studies that report a linkage between HABs and human health, especially along the four major rivers where dams, weirs, and reservoirs were constructed, and sediments were dredged under the Four Major Rivers Project (FMRP) that ended in 2012. The goals of this study were to summarize spatial distribution patterns of HABs and investigate a potential association between HABs and liver diseases. Chlorophyll-a concentration was used to estimate bloom intensity since it was the only available bloom-related parameter that covers the entire rivers. Liver disease data (ICD-10 codes: K71-K77) were sorted by administrative districts. Generalized linear mixed model was used to analyze the bloom, liver diseases, and population data (2005-2016). The results show that chlorophyll-a levels significantly increased since 2013, except Han River region. There was a significant association between HAB intensity and incidence rate of liver diseases, except Han River area, and the extent of association significantly increased after the completion of the FMRP. For future studies, more in-depth epidemiological investigations are warranted in those areas to accurately determine more specific associations between HABs and liver diseases as well as other bloom-related diseases and symptoms. In addition, identification of major exposure pathways to cyanotoxins is needed to better protect public health in those bloom-affected areas.

Modeling outbreak data: Analysis of a 2012 Ebola virus disease epidemic in DRC.

We describe two approaches to modeling data from a small to moderate-sized epidemic outbreak. The first approach is based on a branching process approximation and direct analysis of the transmission network, whereas the second one is based on a survival model derived from the classical SIR equations with no explicit transmission information. We compare these approaches using data from a 2012 outbreak of Ebola virus disease caused by Bundibugyo ebolavirus in city of Isiro, Democratic Republic of the Congo. The branching process model allows for a direct comparison of disease transmission across different environments, such as the general community or the Ebola treatment unit. However, the survival model appears to yield parameter estimates with more accuracy and better precision in some circumstances.

Modeling household transmission dynamics: Application to waterborne diarrheal disease in Central Africa.

INTRODUCTION: We describe a method for analyzing the within-household network dynamics of a disease transmission. We apply it to analyze the occurrences of endemic diarrheal disease in Cameroon, Central Africa based on observational, cross-sectional data available from household health surveys. METHODS: To analyze the data, we apply formalism of the dynamic SID (susceptible-infected-diseased) process that describes the disease steady-state while adjusting for the household age-structure and environment contamination, such as water contamination. The SID transmission rates are estimated via MCMC method with the help of the so-called synthetic likelihood approach. RESULTS: The SID model is fitted to a dataset on diarrhea occurrence from 63 households in Cameroon. We show that the model allows for quantification of the effects of drinking water contamination on both transmission and recovery rates for household diarrheal disease occurrence as well as for estimation of the rate of silent (unobserved) infections. CONCLUSIONS: The new estimation method appears capable of genuinely capturing the complex dynamics of disease transmission across various human, animal and environmental compartments at the household level. Our approach is quite general and can be used in other epidemiological settings where it is desirable to fit transmission rates using cross-sectional data. SOFTWARE SHARING: The R-scripts for carrying out the computational analysis described in the paper are available at https://github.com/cbskust/SID.

Beyond the Michaelis-Menten equation: Accurate and efficient estimation of enzyme kinetic parameters.

Examining enzyme kinetics is critical for understanding cellular systems and for using enzymes in industry. The Michaelis-Menten equation has been widely used for over a century to estimate the enzyme kinetic parameters from reaction progress curves of substrates, which is known as the progress curve assay. However, this canonical approach works in limited conditions, such as when there is a large excess of substrate over enzyme. Even when this condition is satisfied, the identifiability of parameters is not always guaranteed, and often not verifiable in practice. To overcome such limitations of the canonical approach for the progress curve assay, here we propose a Bayesian approach based on an equation derived with the total quasi-steady-state approximation. In contrast to the canonical approach, estimates obtained with this proposed approach exhibit little bias for any combination of enzyme and substrate concentrations. Importantly, unlike the canonical approach, an optimal experiment to identify parameters with certainty can be easily designed without any prior information. Indeed, with this proposed design, the kinetic parameters of diverse enzymes with disparate catalytic efficiencies, such as chymotrypsin, fumarase, and urease, can be accurately and precisely estimated from a minimal amount of timecourse data. A publicly accessible computational package performing such accurate and efficient Bayesian inference for enzyme kinetics is provided.

Estimating epidemic parameters: Application to H1N1 pandemic data.

This paper discusses estimation of the parameters in an SIR epidemic model from the observed longitudinal new infection count data. The potential problems of the standard MLE approaches are revealed and possible remedies suggested. The analysis is based on the epidemic data from the 2009 outbreak of H1N1 influenza on the campus of Washington State University.

Inference for discretely observed stochastic kinetic networks with applications to epidemic modeling.

We present a new method for Bayesian Markov Chain Monte Carlo-based inference in certain types of stochastic models, suitable for modeling noisy epidemic data. We apply the so-called uniformization representation of a Markov process, in order to efficiently generate appropriate conditional distributions in the Gibbs sampler algorithm. The approach is shown to work well in various data-poor settings, that is, when only partial information about the epidemic process is available, as illustrated on the synthetic data from SIR-type epidemics and the Center for Disease Control and Prevention data from the onset of the H1N1 pandemic in the United States.