Effects of Etiological Agent and Bather Shedding of Pathogens on Interpretation of Epidemiological Data Used to Establish Recreational Water Quality Standards

The overall goal of the study reported herein was to use techniques in the field of risk assessment (specifically a state-space population dynamic model of disease transmission within recreational waters) to explore the relative significance of (1) active shedding of microorganisms from bathers themselves, and (2) the type and concentration of etiological agent on the observed heterogeneity of the incidence of illness in epidemiological studies that have been used to develop ambient water quality criteria. The etiological agent and corresponding dose ingested during recreational contact was found to significantly impact the observed incidence of illness in an epidemiological study conducted in recreational water. In addition, the observed incidence of illness was found not to necessarily reflect background concentrations of indicator organisms, but rather microorganisms shed during recreational contact. Future revisions to ambient water quality criteria should address the etiological agent, dose, and the significance of microbial shedding relative to background concentrations of pathogens and indicator organisms in addition to the incidence of illness and concentration of indicator organisms. Without a quantitative assessment of these additional variables, study findings may potentially be site specific and not representative of the health risks associated with specific indicator concentrations in all recreational waters.     

Impact of Acquired Immunity and Dose‐Dependent Probability of Illness on Quantitative Microbial Risk Assessment

Dose‐response models in microbial risk assessment consider two steps in the process ultimately leading to illness: from exposure to (asymptomatic) infection, and from infection to (symptomatic) illness. Most data and theoretical approaches are available for the exposure‐infection step; the infection‐illness step has received less attention. Furthermore, current microbial risk assessment models do not account for acquired immunity. These limitations may lead to biased risk estimates. We consider effects of both dose dependency of the conditional probability of illness given infection, and acquired immunity to risk estimates, and demonstrate their effects in a case study on exposure to Campylobacter jejuni. To account for acquired immunity in risk estimates, an inflation factor is proposed. The inflation factor depends on the relative rates of loss of protection over exposure. The conditional probability of illness given infection is based on a previously published model, accounting for the within‐host dynamics of illness. We find that at low (average) doses, the infection‐illness model has the greatest impact on risk estimates, whereas at higher (average) doses and/or increased exposure frequencies, the acquired immunity model has the greatest impact. The proposed models are strongly nonlinear, and reducing exposure is not expected to lead to a proportional decrease in risk and, under certain conditions, may even lead to an increase in risk. The impact of different dose‐response models on risk estimates is particularly pronounced when introducing heterogeneity in the population exposure distribution.     

Predictive Bayesian Microbial Dose-Response Assessment Based on Suggested Self-Organization in Primary Illness Response: Cryptosporidium parvum

The probability of illness caused by very low doses of pathogens cannot generally be tested due to the numbers of subjects that would be needed, though such assessments of illness dose response are needed to evaluate drinking water standards. A predictive Bayesian dose-response assessment method was proposed previously to assess the unconditional probability of illness from available information and avoid the inconsistencies of confidence-based approaches. However, the method uses knowledge of the conditional dose-response form, and this form is not well established for the illness endpoint. A conditional parametric dose-response function for gastroenteric illness is proposed here based on simple numerical models of self-organized host-pathogen systems and probabilistic arguments. In the models, illnesses terminate when the host evolves by processes of natural selection to a self-organized critical value of wellness. A generalized beta-Poisson illness dose-response form emerges for the population as a whole. Use of this form is demonstrated in a predictive Bayesian dose-response assessment for cryptosporidiosis. Results suggest that a maximum allowable dose of 5.0 x 10(-7) oocysts/exposure (e.g., 2.5 x 10(-7) oocysts/L water) would correspond with the original goals of the U.S. Environmental Protection Agency Surface Water Treatment Rule, considering only primary illnesses resulting from Poisson-distributed pathogen counts. This estimate should be revised to account for non-Poisson distributions of Cryptosporidium parvum in drinking water and total response, considering secondary illness propagation in the population.     

Construction of a Dose–Illness Relationship via Modeling Morbidity and Application to Risk Assessment of Wastewater Reuse

A disease burden (DB) evaluation for environmental pathogens is generally performed using disability‐adjusted life years with the aim of providing a quantitative assessment of the health hazard caused by pathogens. A critical step in the preparation for this evaluation is the estimation of morbidity between exposure and disease occurrence. In this study, the method of a traditional dose–response analysis was first reviewed, and then a combination of the theoretical basis of a “single‐hit” and an “infection‐illness” model was performed by incorporating two critical factors: the “infective coefficient” and “infection duration.” This allowed a dose–morbidity model to be built for direct use in DB calculations. In addition, human experimental data for typical intestinal pathogens were obtained for model validation, and the results indicated that the model was well fitted and could be further used for morbidity estimation. On this basis, a real case of a water reuse project was selected for model application, and the morbidity as well as the DB caused by intestinal pathogens during water reuse was evaluated. The results show that the DB attributed to Enteroviruses was significant, while that for enteric bacteria was negligible. Therefore, water treatment technology should be further improved to reduce the exposure risk of Enteroviruses . Since road flushing was identified as the major exposure route, human contact with reclaimed water through this pathway should be limited. The methodology proposed for model construction not only makes up for missing data of morbidity during risk evaluation, but is also necessary to quantify the maximum possible DB.     

Human Salmonellosis: Estimation of Dose-Illness from Outbreak Data

The quantification of the relationship between the amount of microbial organisms ingested and a specific outcome such as infection, illness, or mortality is a key aspect of quantitative risk assessment. A main problem in determining such dose-response models is the availability of appropriate data. Human feeding trials have been criticized because only young healthy volunteers are selected to participate and low doses, as often occurring in real life, are typically not considered. Epidemiological outbreak data are considered to be more valuable, but are more subject to data uncertainty. In this article, we model the dose-illness relationship based on data of 20 Salmonella outbreaks, as discussed by the World Health Organization. In particular, we model the dose-illness relationship using generalized linear mixed models and fractional polynomials of dose. The fractional polynomial models are modified to satisfy the properties of different types of dose-illness models as proposed by Teunis et al . Within these models, differences in host susceptibility (susceptible versus normal population) are modeled as fixed effects whereas differences in serovar type and food matrix are modeled as random effects. In addition, two bootstrap procedures are presented. A first procedure accounts for stochastic variability whereas a second procedure accounts for both stochastic variability and data uncertainty. The analyses indicate that the susceptible population has a higher probability of illness at low dose levels when the combination pathogen-food matrix is extremely virulent and at high dose levels when the combination is less virulent. Furthermore, the analyses suggest that immunity exists in the normal population but not in the susceptible population.     

An Integrated Model of Infection Risk in a Health-Care Environment

Certain respiratory tract infections can be transmitted by hand-to-mucous-membrane contact, inhalation, and/or direct respiratory droplet spray. In a room occupied by a patient with such a transmissible infection, pathogens present on textile and nontextile surfaces, and pathogens present in the air, provide sources of exposure for an attending health-care worker (HCW); in addition, close contact with the patient when the latter coughs allows for droplet spray exposure. We present an integrated model of pertinent source-environment-receptor pathways, and represent physical elements in these pathways as "states" in a discrete-time Markov chain model. We estimate the rates of transfer at various steps in the pathways, and their relationship to the probability that a pathogen in one state has moved to another state by the end of a specified time interval. Given initial pathogen loads on textile and nontextile surfaces and in room air, we use the model to estimate the expected pathogen dose to a HCW's mucous membranes and respiratory tract. In turn, using a nonthreshold infectious dose model, we relate the expected dose to infection risk. The system is illustrated with a hypothetical but plausible scenario involving a viral pathogen emitted via coughing. We also use the model to show that a biocidal finish on textile surfaces has the potential to substantially reduce infection risk via the hand-to-mucous-membrane exposure pathway.     

Cryptosporidium Dose-Response Studies: Variation Between Hosts

The issue of variation is highly important in dose-response analysis: variation among genetically related pathogens infecting the same host, but also variation among hosts, in susceptibility to infection by the same pathogen. This latter issue is addressed here for the protozoan parasite Cryptosporidium parvum, the causative agent for many outbreaks of water-borne gastrointestinal illness. In human feeding studies, infectivity has been shown to be low in subjects with high preexisting anti-Cryptosporidium IgG-levels. Here we adapt the hit theory model of microbial infection to incorporate covariables, characterizing the immune status of the susceptible host. The probability of any single oocyst in the inoculum to cause infection appears to depend on preexisting IgG-levels. This does not necessarily imply direct protection by the humoral immune system; high IgG-levels may reflect a recent episode of infection/illness, and be an epi-phenomenon associated with other protective responses. The IgG-dependence of the dose-response relation can be easily applied in quantitative risk analysis. The distribution of anti-Cryptosporidium IgG levels in the general population is accessible by analyzing serum banks, which are maintained in many Western countries. Using such an approach provides first insights into the variation of susceptibility to infection in the general population.     

Methodology for Determining the Appropriateness of a Linear Dose‐Response Function

Microbial food safety risk assessment models can often at times be simplified by eliminating the need to integrate a complex dose‐response relationship across a distribution of exposure doses. This is possible if exposure pathways lead to pathogens at exposure that consistently have a small probability of causing illness. In this situation, the probability of illness will follow an approximately linear function of dose. Consequently, the predicted probability of illness per serving across all exposures is linear with respect to the expected value of dose. The majority of dose‐response functions are approximately linear when the dose is low. Nevertheless, what constitutes “low” is dependent on the parameters of the dose‐response function for a particular pathogen. In this study, a method is proposed to determine an upper bound of the exposure distribution for which the use of a linear dose‐response function is acceptable. If this upper bound is substantially larger than the expected value of exposure doses, then a linear approximation for probability of illness is reasonable. If conditions are appropriate for using the linear dose‐response approximation, for example, the expected value for exposure doses is two to three logs₁₀ smaller than the upper bound of the linear portion of the dose‐response function, then predicting the risk‐reducing effectiveness of a proposed policy is trivial. Simple examples illustrate how this approximation can be used to inform policy decisions and improve an analyst's understanding of risk.     

Model Averaging Using the Kullback Information Criterion in Estimating Effective Doses for Microbial Infection and Illness

Since the National Food Safety Initiative of 1997, risk assessment has been an important issue in food safety areas. Microbial risk assessment is a systematic process for describing and quantifying a potential to cause adverse health effects associated with exposure to microorganisms. Various dose-response models for estimating microbial risks have been investigated. We have considered four two-parameter models and four three-parameter models in order to evaluate variability among the models for microbial risk assessment using infectivity and illness data from studies with human volunteers exposed to a variety of microbial pathogens. Model variability is measured in terms of estimated ED01s and ED10s, with the view that these effective dose levels correspond to the lower and upper limits of the 1% to 10% risk range generally recommended for establishing benchmark doses in risk assessment. Parameters of the statistical models are estimated using the maximum likelihood method. In this article a weighted average of effective dose estimates from eight two- and three-parameter dose-response models, with weights determined by the Kullback information criterion, is proposed to address model uncertainties in microbial risk assessment. The proposed procedures for incorporating model uncertainties and making inferences are illustrated with human infection/illness dose-response data sets.     

Dose Response for Infectivity of Several Strains of Campylobacter jejuni in Chickens

Although some major risk studies have been done for Campylobacter jejuni, its dose response is not well characterized. Only a single human study is available, providing dose-response information for only a single isolate. As substantial heterogeneity in infectivity has been acknowledged for other pathogens, it remains unknown how well this single study represents the dose-response relation for this pathogen. As future human challenge studies with Campylobacter are unlikely, we have to find other means of studying its infectivity. Several dose-response studies have been done using chickens as host organisms. These studies may be used to obtain quantitative information on the variation in infectivity among different isolates of this pathogen. A hierarchical Bayesian model is well suited to describe heterogeneity, and we demonstrate how the beta-Poisson model of microbial infection may be adapted to allow for within- and between-isolate variation. Isolates tested in chickens can be categorized into two distinct groups: lab-adapted and fresh isolates, and we show how the hierarchical dose-response model can be used to quantitatively describe their differences. Fresh isolates show higher colonization potential and less within-isolate variation than lab isolates. The results indicate that Campylobacter jejuni is highly infectious in chickens. Different isolates show great variation in infectivity, especially between lab and fresh isolates, indicating that human clinical (volunteer) studies on infectivity must be interpreted cautiously.     

Microbiota and Dose Response: Evolving Paradigm of Health Triangle

SRA Dose‐Response and Microbial Risk Analysis Specialty Groups jointly sponsored symposia that addressed the intersections between the “microbiome revolution” and dose response. Invited speakers presented on innovations and advances in gut and nasal microbiota (normal microbial communities) in the first decade after the Human Microbiome Project began. The microbiota and their metabolites are now known to influence health and disease directly and indirectly, through modulation of innate and adaptive immune systems and barrier function. Disruption of healthy microbiota is often associated with changes in abundance and diversity of core microbial species (dysbiosis), caused by stressors including antibiotics, chemotherapy, and disease. Nucleic‐acid‐based metagenomic methods demonstrated that the dysbiotic host microbiota no longer provide normal colonization resistance to pathogens, a critical component of innate immunity of the superorganism. Diverse pathogens, probiotics, and prebiotics were considered in human and animal models (in vivo and in vitro ). Discussion included approaches for design of future microbial dose–response studies to account for the presence of the indigenous microbiota that provide normal colonization resistance , and the absence of the protective microbiota in dysbiosis. As NextGen risk analysis methodology advances with the “microbiome revolution,” a proposed new framework, the Health Triangle, may replace the old paradigm based on the Disease Triangle (focused on host, pathogen, and environment) and germophobia. Collaborative experimental designs are needed for testing hypotheses about causality in dose–response relationships for pathogens present in our environments that clearly compete in complex ecosystems with thousands of bacterial species dominating the healthy superorganism.     

Model Uncertainty and Model Averaging in the Estimation of Infectious Doses for Microbial Pathogens

Food‐borne infection is caused by intake of foods or beverages contaminated with microbial pathogens. Dose‐response modeling is used to estimate exposure levels of pathogens associated with specific risks of infection or illness. When a single dose‐response model is used and confidence limits on infectious doses are calculated, only data uncertainty is captured. We propose a method to estimate the lower confidence limit on an infectious dose by including model uncertainty and separating it from data uncertainty. The infectious dose is estimated by a weighted average of effective dose estimates from a set of dose‐response models via a Kullback information criterion. The confidence interval for the infectious dose is constructed by the delta method, where data uncertainty is addressed by a bootstrap method. To evaluate the actual coverage probabilities of the lower confidence limit, a Monte Carlo simulation study is conducted under sublinear, linear, and superlinear dose‐response shapes that can be commonly found in real data sets. Our model‐averaging method achieves coverage close to nominal in almost all cases, thus providing a useful and efficient tool for accurate calculation of lower confidence limits on infectious doses.     

A Risk Analysis for Airborne Pathogens with Low Infectious Doses: Application to Respirator Selection Against Coccidioides immitis Spores

Probability models incorporating a deterministic versus stochastic infectious dose are described for estimating infection risk due to airborne pathogens that infect at low doses. Such pathogens can be occupational hazards or candidate agents for bioterrorism. Inputs include parameters for the infectious dose model, distribution parameters for ambient pathogen concentrations, the breathing rate, the duration of an exposure period, the anticipated number of exposure periods, and, if a respirator device is used, distribution parameters for respirator penetration values. Application of the models is illustrated with a hypothetical scenario involving exposure to Coccidioides immitis, a fungus present in soil in areas of the southwestern United States Inhaling C. immitis spores causes a respiratory tract infection and is a recognized occupational hazard in jobs involving soil dust exposure in endemic areas An uncertainty analysis is applied to risk estimation in the context of selecting respiratory protection with a desired degree of efficacy.     

QMRA for Drinking Water: 1. Revisiting the Mathematical Structure of Single‐Hit Dose‐Response Models

Dose‐response models are essential to quantitative microbial risk assessment (QMRA), providing a link between levels of human exposure to pathogens and the probability of negative health outcomes. In drinking water studies, the class of semi‐mechanistic models known as single‐hit models, such as the exponential and the exact beta‐Poisson, has seen widespread use. In this work, an attempt is made to carefully develop the general mathematical single‐hit framework while explicitly accounting for variation in (1) host susceptibility and (2) pathogen infectivity. This allows a precise interpretation of the so‐called single‐hit probability and precise identification of a set of statistical independence assumptions that are sufficient to arrive at single‐hit models. Further analysis of the model framework is facilitated by formulating the single‐hit models compactly using probability generating and moment generating functions. Among the more practically relevant conclusions drawn are: (1) for any dose distribution, variation in host susceptibility always reduces the single‐hit risk compared to a constant host susceptibility (assuming equal mean susceptibilities), (2) the model‐consistent representation of complete host immunity is formally demonstrated to be a simple scaling of the response, (3) the model‐consistent expression for the total risk from repeated exposures deviates (gives lower risk) from the conventional expression used in applications, and (4) a model‐consistent expression for the mean per‐exposure dose that produces the correct total risk from repeated exposures is developed.     

Fractional Poisson—A Simple Dose‐Response Model for Human Norovirus

This study utilizes old and new Norovirus (NoV) human challenge data to model the dose‐response relationship for human NoV infection. The combined data set is used to update estimates from a previously published beta‐Poisson dose‐response model that includes parameters for virus aggregation and for a beta‐distribution that describes variable susceptibility among hosts. The quality of the beta‐Poisson model is examined and a simpler model is proposed. The new model (fractional Poisson) characterizes hosts as either perfectly susceptible or perfectly immune, requiring a single parameter (the fraction of perfectly susceptible hosts) in place of the two‐parameter beta‐distribution. A second parameter is included to account for virus aggregation in the same fashion as it is added to the beta‐Poisson model. Infection probability is simply the product of the probability of nonzero exposure (at least one virus or aggregate is ingested) and the fraction of susceptible hosts. The model is computationally simple and appears to be well suited to the data from the NoV human challenge studies. The model's deviance is similar to that of the beta‐Poisson, but with one parameter, rather than two. As a result, the Akaike information criterion favors the fractional Poisson over the beta‐Poisson model. At low, environmentally relevant exposure levels (<100), estimation error is small for the fractional Poisson model; however, caution is advised because no subjects were challenged at such a low dose. New low‐dose data would be of great value to further clarify the NoV dose‐response relationship and to support improved risk assessment for environmentally relevant exposures.     

Analysis of Regional Scale Risk of Whirling Disease in Populations of Colorado and Rio Grande Cutthroat Trout Using a Bayesian Belief Network Model

Introduction and spread of the parasite Myxobolus cerebralis, the causative agent of whirling disease, has contributed to the collapse of wild trout populations throughout the intermountain west. Of concern is the risk the disease may have on conservation and recovery of native cutthroat trout. We employed a Bayesian belief network to assess probability of whirling disease in Colorado River and Rio Grande cutthroat trout (Oncorhynchus clarkii pleuriticus and Oncorhynchus clarkii virginalis, respectively) within their current ranges in the southwest United States. Available habitat (as defined by gradient and elevation) for intermediate oligochaete worm host, Tubifex tubifex, exerted the greatest influence on the likelihood of infection, yet prevalence of stream barriers also affected the risk outcome. Management areas that had the highest likelihood of infected Colorado River cutthroat trout were in the eastern portion of their range, although the probability of infection was highest for populations in the southern, San Juan subbasin. Rio Grande cutthroat trout had a relatively low likelihood of infection, with populations in the southernmost Pecos management area predicted to be at greatest risk. The Bayesian risk assessment model predicted the likelihood of whirling disease infection from its principal transmission vector, fish movement, and suggested that barriers may be effective in reducing risk of exposure to native trout populations. Data gaps, especially with regard to location of spawning, highlighted the importance in developing monitoring plans that support future risk assessments and adaptive management for subspecies of cutthroat trout.     

Quantitative Risk Assessment of Vibrio parahaemolyticus in Finfish: A Model of Raw Horse Mackerel Consumption in Japan

The aim of this study was to evaluate the effects of implemented control measures to reduce illness induced by Vibrio parahaemolyticus (V. parahaemolyticus) in horse mackerel (Trachurus japonicus), seafood that is commonly consumed raw in Japan. On the basis of currently available experimental and survey data, we constructed a quantitative risk model of V. parahaemolyticus in horse mackerel from harvest to consumption. In particular, the following factors were evaluated: bacterial growth at all stages, effects of washing the fish body and storage water, and bacterial transfer from the fish surface, gills, and intestine to fillets during preparation. New parameters of the beta‐Poisson dose‐response model were determined from all human feeding trials, some of which have been used for risk assessment by the U.S. Food and Drug Administration (USFDA). The probability of illness caused by V. parahaemolyticus was estimated using both the USFDA dose‐response parameters and our parameters for each selected pathway of scenario alternatives: washing whole fish at landing, storage in contaminated water, high temperature during transportation, and washing fish during preparation. The last scenario (washing fish during preparation) was the most effective for reducing the risk of illness by about a factor of 10 compared to no washing at this stage. Risk of illness increased by 50% by exposure to increased temperature during transportation, according to our assumptions of duration and temperature. The other two scenarios did not significantly affect risk. The choice of dose‐response parameters was not critical for evaluation of control measures.     

A Quantitative Risk Assessment of Waterborne Cryptosporidiosis in France Using Second-Order Monte Carlo Simulation

A pragmatic quantitative risk assessment (QRA) of the risks of waterborne Cryptosporidium parvum infection and cryptosporidiosis in immunocompetent and immunodeficient French populations is proposed. The model takes into account French specificities such as the French technique for oocyst enumeration performance and tap water consumption. The proportion of infective oocysts is based on literature review and expert knowledge. The probability of infection for a given number of ingested viable oocysts is modeled using the exponential dose-response model applied on published data from experimental infections in immunocompetent human volunteers challenged with the IOWA strain. Second-order Monte Carlo simulations are used to characterize the uncertainty and variability of the risk estimates. Daily risk of infection and illness for the immunocompetent and the immunodeficient populations are estimated according to the number of oocysts observed in a single storage reservoir water sample. As an example, the mean daily risk of infection in the immunocompetent population is estimated to be 1.08 x 10(-4) (95% confidence interval: [0.20 x 10(-4); 6.83 x 10(-4)]) when five oocysts are observed in a 100 L storage reservoir water sample. Annual risks of infection and disease are estimated from a set of oocyst enumeration results from distributed water samples, assuming a negative binomial distribution of day-to-day contamination variation. The model and various assumptions used in the model are fully explained and discussed. While caveats of this model are well recognized, this pragmatic QRA could represent a useful tool for the French Food Safety Agency (AFSSA) to define recommendations in case of water resource contamination by C. parvum whose infectivity is comparable to the IOWA strain.     

Dose Response for Infection by Escherichia coli O157:H7 from Outbreak Data

In 1996, an outbreak of E. coli O157:H7-associated illness occurred in an elementary school in Japan. This outbreak has been studied in unusual detail, making this an important case for quantitative risk assessment. The availability of stored samples of the contaminated food allowed reliable estimation of exposure to the pathogens. Collection of fecal samples allowed assessment of the numbers infected, including asymptomatic cases. Comparison to other published dose-response studies for E. coli O157:H7 show that the strain that caused the outbreak studied here must have been considerably more infectious. We use this well-documented incident as an example to demonstrate how such information on the response to a single dose can be used for dose-response assessment. In particular, we demonstrate how the high infectivity limits the uncertainty in the low-dose region.     

Norovirus Dose–Response: Are Currently Available Data Informative Enough to Determine How Susceptible Humans Are to Infection from a Single Virus?

Two forms of single‐hit infection dose‐response models have previously been developed to assess available data from human feeding trials and estimate the norovirus dose‐response relationship. The mechanistic interpretations of these models include strong assumptions that warrant reconsideration: the first study includes an implicit assumption that there is no immunity to Norwalk virus among the specific study population, while the recent second study includes assumptions that such immunity could exist and that the nonimmune have no defensive barriers to prevent infection from exposure to just one virus. Both models addressed unmeasured virus aggregation in administered doses. In this work, the available data are reanalyzed using a generalization of the first model to explore these previous assumptions. It was hypothesized that concurrent estimation of an unmeasured degree of virus aggregation and important dose‐response parameters could lead to structural nonidentifiability of the model (i.e., that a diverse range of alternative mechanistic interpretations yield the same optimal fit), and this is demonstrated using the profile likelihood approach and by algebraic proof. It is also demonstrated that omission of an immunity parameter can artificially inflate the estimated degree of aggregation and falsely suggest high susceptibility among the nonimmune. The currently available data support the assumption of immunity within the specific study population, but provide only weak information about the degree of aggregation and susceptibility among the nonimmune. The probability of infection at low and moderate doses may be much lower than previously asserted, but more data from strategically designed dose‐response experiments are needed to provide adequate information.