Rebecca L. Calderon

Norwalk virus: How infectious is it?†

Noroviruses are major agents of viral gastroenteritis worldwide. The infectivity of Norwalk virus, the prototype norovirus, has been studied in susceptible human volunteers. A new variant of the hit theory model of microbial infection was developed to estimate the variation in Norwalk virus infectivity, as well as the degree of virus aggregation, consistent with independent (electron microscopic) observations. Explicit modeling of viral aggregation allows us to express virus infectivity per single infectious unit (particle). Comparison of a primary and a secondary inoculum showed that passage through a human host does not change Norwalk virus infectivity. We estimate the average probability of infection for a single Norwalk virus particle to be close to 0.5, exceeding that reported for any other virus studied to date. Infected subjects had a dose‐dependent probability of becoming ill, ranging from 0.1 (at a dose of 103 NV genomes) to 0.7 (at 108 virus genomes). A norovirus dose response model is important for understanding its transmission and essential for development of a quantitative risk model. Norwalk virus is a valuable model system to study virulence because genetic factors are known for both complete and partial protection; the latter can be quantitatively described as heterogeneity in dose response models. J. Med. Virol. 80:1468–1476, 2008.

Rapidly measured indicators of recreational water quality are predictive of swimming-associated gastrointestinal illness.

Standard methods to measure recreational water quality require at least 24 hr to obtain results, making it impossible to assess the quality of water within a single day. Methods to measure recreational water quality in ≤ 2 hr have been developed. Application of rapid methods could give considerably more accurate and timely assessments of recreational water quality. We conducted a prospective study of beachgoers at two Great Lakes beaches to examine the association between recreational water quality, obtained using rapid methods, and gastrointestinal (GI) illness after swimming. Beachgoers were asked about swimming and other beach activities and 10–12 days later were asked about the occurrence of GI symptoms. We tested water samples for Enterococcus and Bacteroides species using the quantitative polymerase chain reaction (PCR) method. We observed significant trends between increased GI illness and Enterococcus at the Lake Michigan beach and a positive trend for Enterococcus at the Lake Erie beach. The association remained significant for Enterococcus when the two beaches were combined. We observed a positive trend for Bacteroides at the Lake Erie beach, but no trend was observed at the Lake Michigan beach. Enterococcus samples collected at 0800 hr were predictive of GI illness that day. The association between Enterococcus and illness strengthened as time spent swimming in the water increased. This is the first study to show that water quality measured by rapid methods can predict swimming-associated health effects.

High sensitivity of children to swimming-associated gastrointestinal illness: results using a rapid assay of recreational water quality.

Background: Culture-based methods of monitoring fecal pollution in recreational waters require 24 to 48 hours to obtain results. This delay leads to potentially inaccurate management decisions regarding beach safety. We evaluated the quantitative polymerase chain reaction (QPCR) as a faster method to assess recreational water quality and predict swimming-associated illnesses. Methods: We enrolled visitors at 4 freshwater Great Lakes beaches, and contacted them 10 to 12 days later to ask about health symptoms experienced since the visit. Water at the beaches was polluted by point sources that carried treated sewage. We tested water samples daily for Enterococcus using QPCR and membrane filtration (EPA Method 1600). Results: We completed 21,015 interviews and tested 1359 water samples. Enterococcus QPCR cell equivalents (CEs) were positively associated with swimming-associated gastrointestinal (GI) illness (adjusted odds ratio per 1 log10 QPCR CE =1.26; 95% confidence interval = 1.06–1.51). The association between GI illness and QPCR CE was stronger among children aged 10 years and below (1.69; 1.24–2.30). Nonenteric illnesses were not consistently associated with Enterococcus QPCR CE exposure, although rash and earache occurred more frequently among swimmers. Enterococcus QPCR CE exposure was more strongly associated with GI illness than Enterococcus measured by membrane filtration. Conclusions: Measurement of the indicator bacteria Enterococci in recreational water using a rapid QPCR method predicted swimming-associated GI illness at freshwater beaches polluted by sewage discharge. Children at 10 years or younger were at greater risk for GI illness following exposure.

Serological evidence of endemic waterborne cryptosporidium infections.

PURPOSE Cryptosporidium oocysts are commonly detected in surface-derived drinking water, however, the public health significance of these findings is unclear. This study compared the evidence of prior Cryptosporidium infection for people drinking water derived from surface versus ground water sources. METHODS This study measured serological responses to two Cryptosporidium antigen groups for blood donors from two midwestern United States cities with different drinking water sources: filtered and chlorinated river water receiving agricultural and domestic sewage upstream versus chlorinated water from an underground aquifer. Initial and nine-month paired serological responses to two Cryptosporidium antigen groups were compared. RESULTS Initially, donors from the surface water city had a higher relative prevalence (RP) of a serological response (54% vs. 38%, RP = 1.39 (1.21,1.60)). Donors with a detectable baseline response who resided in the surface water city had a higher relative risk (RR) of an increased intensity of response on the follow-up blood draw (15/17-kDa, 40/100 vs. 11/100, RR = 3.78 (1.89,7.58)), (27-kDa 38/100 vs. 18/100, RR = 2.07 (1.31,3.25)). Donors with no baseline response to the 15/17-kDa marker who resided in the surface water city also had a higher risk of serconversion (38/100 vs. 14/100, RR = 2.63 (1.78,3.91)). CONCLUSIONS These rates of both background and drinking water related Cryptosporidium infections are substantially higher than previously estimated, however, the risk of illness from infection may be lower.

Serological evidence of Cryptosporidium infections in southern Europe

Although cryptosporidiosis outbreaks have been frequently reported in the United States, Canada and the United Kingdom, few outbreaks have been reported on the European continent. The reasons for this are unclear. To ascertain whether a European population has been previously exposed to Cryptosporidium, we conducted a survey of 100 resident blood donors in a northern Italian city for IgG serological response to two oocyst antigen groups. A serological response to the 15/17-kDa antigen group was detected in 83% of blood donors and response to the 27-kDa antigen group in 62%. Donors who traveled outside of Italy during the prior 12 months were less likely to have had a response to the 15/17-kDa antigen group (p < 0.04) and to have a less intense response (p < 0.05). Older age was predictive of a more intense response to each antigen group (p < 0.01). The fraction of Italian blood donors with a serological response to either antigen group was higher than in four United States blood donor populations, with differences more pronounced for response to the 15/17-kDa antigen group (p < 0.01). A lower fraction of Italian donors had a serological response to either antigen group than persons tested at the time of a cryptosporidiosis outbreak in the United States or blood donors tested six months after that outbreak (p < 0.05). Since the presence of serological responses to these antigen groups predicts a reduced risk of cryptosporidiosis, the high prevalence of serological responses in these Italian blood donors may explain the infrequent occurrences of clinically detectable cryptosporidiosis in this city.

Biological and behavioral factors modify biomarkers of arsenic exposure in a U.S. population

Although consumption of drinking water contaminated with inorganic arsenic is usually considered the primary exposure route, aggregate exposure to arsenic depends on direct consumption of water, use of water in food preparation, and the presence in arsenicals in foods. To gain insight into the effects of biological and behavioral factors on arsenic exposure, we determined arsenic concentrations in urine and toenails in a U.S. population that uses public or private water supplies containing inorganic arsenic. Study participants were 904 adult residents of Churchill County, Nevada, whose home tap water supplies contained <3 to about 1200 µg of arsenic per liter. Biomarkers of exposure for this study were summed urinary concentrations of inorganic arsenic and its methylated metabolites (speciated arsenical), of all urinary arsenicals (total arsenical), and of all toenail arsenicals (total arsenical). Increased tap water arsenic concentration and consumption were associated with significant upward trends for urinary speciated and total and toenail total arsenical concentrations. Significant gender differences in concentrations of speciated and total arsenicals in urine and toenails reflected male-female difference in water intake. Both recent and higher habitual seafood consumption significantly increased urinary total but not speciated arsenical concentration. In a stepwise general linear model, seafood consumption significantly predicted urinary total arsenical but not urinary speciated or toenail total arsenical concentrations. Smoking behavior significantly predicted urinary speciated or total arsenical concentration. Gender, tap water arsenic concentration, and primary drinking water source significantly predicted urinary speciated and total concentrations and toenail total arsenical concentrations. These findings confirm the primacy of home tap water as a determinant of arsenic concentration in urine and toenails. However, biological and behavioral factors can modify exposure-response relations for these biomarkers. Refining estimates of the influence of these factors will permit better models of dose-response relations for this important environmental contaminant.

Improving waterborne disease outbreak investigations

This article is a summary of discussions held and recommendations made at a workshop for the investigation of waterborne disease outbreaks in Chapel Hill, North Carolina, December 7‐8, 1998. Suspected waterborne outbreaks in the United States are primarily investigated by state and local public health officials who may infrequently conduct enteric disease outbreak investigations. Thus, it is important that officials have a formal plan to ensure that epidemiological studies are methodologically sound and that effective collaboration occurs among the epidemiologists, scientists, and engineers who will conduct the investigations. Laboratory support to analyze water samples and clinical specimens should be arranged well in advance of when services may be needed. Enhanced surveillance activities can help officials recognize additional outbreaks and initiate investigations in a timely manner. Epidemiologists should pay more attention early in the investigation to study design, questionnaire development, and sources of bias, especially recall bias, that may affect the interpretation of observed associations. Improved investigations can increase our knowledge about important etiological agents, water systems deficiencies, and sources of water contamination so that waterborne outbreaks can be more effectively prevented.

Paired city cryptosporidium serosurvey in the southwest USA.

In 1996, serological responses to two Cryptosporidium antigens were determined for 200 Las Vegas (LV), Nevada, and 200 Albuquerque, New Mexico, blood donors to evaluate associations between endemic infections, water exposures, and other risk factors. LV uses chlorinated filtered drinking water from Lake Mead while Albuquerque uses chlorinated ground water. The intensity of serological response to both markers was higher for older donors (P < 0.05). donors who washed food with bottled water (P < 0.05) and donors from LV (P < 0.05). A decreased serological response was not associated with bottled water consumption, nor was an increased response associated with self-reported cryptosporidiosis-like illness or residence in LV at the time of a cryptosporidiosis outbreak 2 years earlier. Although these findings suggest the serological response may be associated with type of tap water and certain foods, additional research is needed to clarifythe role of both food and drinking water in endemic Cryptosporidium infection.

Serological responses to Cryptosporidium antigens among users of surface- vs. ground-water sources

Cryptosporidium oocysts are commonly detected in surface-derived drinking water. However, the public health significance of these findings is unclear. This study compared serological responses to two Cryptosporidium antigen groups for blood donors and college students using chlorinated and filtered river water vs. ground-water sources. The surface water received agricultural and domestic sewage discharges upstream. Participants from the surface-water city had a higher relative prevalence (RP) of a serological response to the 15/17-kDa antigen group (72.3 vs. 52.4%, RP = 1.36, P < 0.001) and to the 27-kDa antigen group (82.6 vs. 72.5%, RP = 1.14, P < 0.02). Multivariate logistic regression analysis found that the people with a shorter duration of residence or drinking bottled water also had a lower seropositivity for each marker. Use of private wells was associated with a higher prevalence of response to the 15/17-kDa markers. Seroconversion to the 15/17-kDa antigen group was more common in the residents of the city using surface water. These findings are consistent with an increased risk of Cryptosporidium infection for users of surface-derived drinking water compared with users of municipal ground-water-derived drinking water. Users of private well water may also have an increased risk of infection.

Consequences of acute and chronic exposure to arsenic in children.

Arsenic is a toxic chemical and may cause adverse health effects in children and adults. It is known to affect the nervous, gastrointestinal, and hematological systems and cause skin and internal cancers in people exposed to levels greater than 300 ppb in their drinking water. For most people, the major exposure to arsenic comes from food (8 to 14 microg inorganic arsenic per day), but when the arsenic level in water is elevated, drinking water becomes the predominant source of exposure. Because it is very difficult to limit arsenic exposure from food, it would be wise to limit arsenic exposure from those more controllable sources. Pediatricians should ascertain the levels of arsenic in drinking water of patients with high arsenic levels, using the supplier or, in the case of private wells, a professional water-testing laboratory assay. The Safe Drinking Water Act does not cover private wells or those water systems with less than 15 hook-ups or those that serve less than 25 people. Pediatricians should instruct parents to use prepared baby formulas or prepare them using water with the arsenic removed and to curtail playing time for younger children in places that have sand containing large amounts of arsenic. Such procedures will limit arsenic exposure to a minimum.