Responses to pathogen exposure in sentinel juvenile fall-run Chinook salmon in the Sacramento River, CA.

This study investigated how the deployment of juvenile Chinook salmon in ambient river conditions and the subsequent exposure to and infection by pathogens was associated with the changes in the expression of genes involved in immune system functioning, general stress and host development. Juvenile fish were deployed in sentinel cages for 21 days in the Sacramento River, CA, USA. Gill, kidney and intestinal tissue were sampled at 0, 7, 14 and 21 days post-deployment. Pathogen detection and host response were assessed by a combination of molecular and histopathological evaluation. Our findings showed that fish became infected by the parasites Ceratonova shasta, Parvicapsula minibicornis and Ichthyophthirius multifiliis, and to a lesser extent, the bacteria Flavobacterium columnare and Rickettsia-like organisms. Co-infection was common among sentinel fish. Expression of investigated genes was altered following deployment and was often associated with pathogen abundance. This study provides a foundation for future avenues of research investigating pathogens that affect out-migrating Chinook salmon in the Sacramento River, and offers crucial knowledge related to conservation efforts.

Waterborne exposure during non-consumptive domestic use of surface water: a population study across WASH service levels in rural India.

Exposure to pathogens from domestic use of surface water is understudied. In many low- and middle-income countries, surface water is used for hygiene, sanitation, amenity, and recreational purposes. In this study, self-reported use of and structured observations at community ponds were collected to measure waterborne exposure across water and sanitation service levels in a rural population of Khorda District, India. Overall, 86% of households (n = 200) reported using ponds on a regular basis. Among observed people (n = 765), 82% put water into their mouth at least once, with a median frequency of five occurrences per visit. Reported and observation data were combined to estimate the proportion (p) of the population that put water in their mouth at least once per day, and their mean daily rate of oral exposure (OE). These were highest for individuals with neither safely managed water nor basic sanitation access (p = 93%, OE = 14 day-1), but still high among those with both (p = 67%, OE = 6 day-1). The results suggest widespread exposure to waterborne pathogens in settings where non-potable surface water bodies continue to be used for domestic purposes, even among households with access to safely managed drinking water.

Associations between faecal pathogen detection, E. coli concentrations and youth exhibitor biosecurity practices at California county fairs.

Interactions with livestock in public settings such as county and state fairs can expose people and other livestock to faecal material capable of spreading zoonotic enteric pathogens. The goal of this study was to understand these risks by screening livestock faeces (n = 245) and livestock bedding (n = 155) for common zoonotic pathogens (Giardia, Cryptosporidium, Salmonella and Campylobacter spp.) and by measuring faecal indicator, Escherichia coli, concentrations in drinking water (n = 153), feed containers (n = 124) and bedding material (n = 157) in four livestock species (cattle, sheep, goats and swine) from county fairs in California, USA. Results indicated that sheep were most likely to have pathogens detected in faeces and that Giardia was the most frequently detected pathogen in both faeces (11%) and bedding (21%) across all livestock species. Additionally, increasing the number of animals in a holding pen at fairs, increasing the stocking density of animals in transport trailers to fairs, and having access to water in transport trailers significantly increased the odds of detecting pathogens in livestock faeces of any animal species. Observing solid material in water, stale feed and soiled bedding was associated with detecting higher E. coli concentrations. These findings provide evidence of faecal pathogens present at county fairs and suggest that site observations can aid in assessing levels of faecal exposure. The findings also indicate that the use of biosecurity measures such as (a) routine changing of livestock drinking water, feed and bedding, (b) not overstocking animals in holding pens and trailers and (c) keeping species in separate holding areas may reduce the risk of humans and livestock being exposed to faecal pathogens.

Giardia Infection and Trypanosoma Cruzi Exposure in Dogs in the Bosawás Biosphere Reserve, Nicaragua.

Indigenous Mayangna and Miskitu inhabit Nicaragua's remote Bosawás Biosphere Reserve, located in the North Caribbean Coast Autonomous Region. They are sedentary horticulturists who supplement their diet with wild game, hunting with the assistance of dogs. To test whether hunting dogs increased the risk of human exposure to protozoal zoonotic neglected tropical diseases (NTDs), we sampled dogs from three communities varying in population size and level of contact with other communities. We screened dog feces (n = 58) for Giardia and Cryptosporidium DNA and sera (n = 78) for Trypanosoma cruzi antibodies and DNA. Giardia DNA was detected in 22% (13/58) of samples; sequencing revealed the presence of both zoonotic genotypes (assemblages A and B) and dog-specific genotypes (assemblages C and D). Giardia shedding was associated with community and age. Older dogs and those in the two, more accessible communities had greater odds of shedding parasites. Seroprevalence of T. cruzi antibodies, indicating prior exposure, was 9% (7/78). These results contribute to the limited literature on NTDs in indigenous populations, and suggest hunting dogs can both serve as sentinels of environmental NTDs and pose zoonotic risk for their owners and communities.

Estimating Cryptosporidium and Giardia disease burdens for children drinking untreated groundwater in a rural population in India.

BACKGROUND: In many low-income settings, despite improvements in sanitation and hygiene, groundwater sources used for drinking may be contaminated with enteric pathogens such as Cryptosporidium and Giardia, which remain important causes of childhood morbidity. In this study, we examined the contribution of diarrhea caused by Cryptosporidium and Giardia found in groundwater sources used for drinking to the total burden of diarrheal disease among children < 5 in rural India. METHODOLOGY/PRINCIPAL FINDINGS: We studied a population of 3,385 children < 5 years of age in 100 communities of Puri District, Odisha, India. We developed a coupled quantitative microbial risk assessment (QMRA) and susceptible-infected-recovered (SIR) population model based on observed levels of Cryptosporidium and Giardia in improved groundwater sources used for drinking and compared the QMRA-SIR estimates with independently measured all-cause (i.e., all fecal-oral enteric pathogens and exposure pathways) child diarrhea prevalence rates observed in the study population during two monsoon seasons (2012 and 2013). We used site specific and regional studies to inform assumptions about the human pathogenicity of the Cryptosporidium and Giardia species present in local groundwater. In all three human pathogenicity scenarios evaluated, the mean daily risk of Cryptosporidium or Giardia infection (0.06-1.53%), far exceeded the tolerable daily risk of infection from drinking water in the US (< 0.0001%). Depending on which protozoa species were present, median estimates of daily child diarrhea prevalence due to either Cryptosporidium or Giardia infection from drinking water was as high as 6.5% or as low as < 1% and accounted for at least 2.9% and as much as 65.8% of the all-cause diarrhea disease burden measured in children < 5 during the study period. Cryptosporidium tended to account for a greater share of estimated waterborne protozoa infections causing diarrhea than did Giardia. Diarrhea prevalence estimates for waterborne Cryptosporidium infection appeared to be most sensitive to assumptions about the probability of infection from ingesting a single parasite (i.e. the rate parameter in dose-response model), while Giardia infection was most sensitive to assumptions about the viability of parasites detected in groundwater samples. CONCLUSIONS/SIGNIFICANCE: Protozoa in groundwater drinking sources in rural India, even at low concentrations, especially for Cryptosporidium, may account for a significant portion of child diarrhea morbidity in settings were tubewells are used for drinking water and should be more systematically monitored. Preventing diarrheal disease burdens in Puri District and similar settings will benefit from ensuring water is microbiologically safe for consumption and consistent and effective household water treatment is practiced.

Modeling Cryptosporidium and Giardia in Ground and Surface Water Sources in Rural India: Associations with Latrines, Livestock, Damaged Wells, and Rainfall Patterns.

Surface and groundwater contamination with fecal pathogens is a public health concern especially in low-income settings where these sources are used untreated. We modeled observed Cryptosporidium and Giardia contamination in community ponds (n = 94; 79% contaminated), deep tubewells (DTWs) (n = 107; 17%), and shallow tubewells (STWs) (n = 96; 19%) during the 2012 and 2013 monsoon seasons (June-August) in 60 villages in Puri District, India to understand sources and processes of contamination. Detection of Cryptosporidium and/or Giardia in a tubewell was positively associated with damage to the well pad for DTWs, the amount of human loading into pour-flush latrine pits nearby (≤15 m) for STWs, and the village literacy rate (for Giardia in STWs). Pond concentration levels were positively associated with the number of people practicing open defecation within 50 m and the sheep population for Cryptosporidium, and with the village illiteracy rate for Giardia. Recent rainfall increased the risk of Cryptosporidium in STWs (an extreme event) and ponds (any), while increasing seasonal rainfall decreased the risk of Giardia in STWs and ponds. Full latrine coverage in this setting is expected to marginally reduce pond Cryptosporidium contamination (16%) while increasing local groundwater protozoal contamination (87-306%), with the largest increases predicted for Cryptosporidium in STWs.

Human fecal and pathogen exposure pathways in rural Indian villages and the effect of increased latrine coverage.

Efforts to eradicate open defecation and improve sanitation access are unlikely to achieve health benefits unless interventions reduce microbial exposures. This study assessed human fecal contamination and pathogen exposures in rural India, and the effect of increased sanitation coverage on contamination and exposure rates. In a cross-sectional study of 60 villages of a cluster-randomized controlled sanitation trial in Odisha, India, human and domestic animal fecal contamination was measured in community tubewells and ponds (n = 301) and via exposure pathways in homes (n = 354), using Bacteroidales microbial source tracking fecal markers validated in India. Community water sources were further tested for diarrheal pathogens (rotavirus, adenovirus and Vibrio cholerae by quantitative PCR; pathogenic Escherichia coli by multiplex PCR; Cryptosporidium and Giardia by immunomagnetic separation and direct fluorescent antibody microscopy). Exposure pathways in intervention and control villages were compared and relationships with child diarrhea examined. Human fecal markers were rarely detected in tubewells (2.4%, 95%CI: 0.3-4.5%) and ponds (5.6%, 95%CI: 0.8-10.3%), compared to homes (35.4%, 95%CI: 30.4-40.4%). In tubewells, V. cholerae was the most frequently detected pathogen (19.8%, 95%CI: 14.4-25.2%), followed by Giardia (14.8%, 95%CI: 10.0-19.7%). In ponds, Giardia was most often detected (74.5%, 95%CI: 65.7-83.3%), followed by pathogenic E. coli (48.1%, 95%CI: 34.8-61.5%) and rotavirus (44.4%, 95%CI: 34.2-54.7%). At village-level, prevalence of fecal pathogen detection in community drinking water sources was associated with elevated prevalence of child diarrhea within 6 weeks of testing (RR 2.13, 95%CI: 1.25-3.63) while within homes, higher levels of human and animal fecal marker detection were associated with increased risks of subsequent child diarrhea (P = 0.044 and 0.013, respectively). There was no evidence that the intervention, which increased functional latrine coverage and use by 27 percentage points, reduced human fecal contamination in any tested pathway, nor the prevalence of pathogens in water sources. In conclusion, the study demonstrates that (1) improved sanitation alone may be insufficient and further interventions needed in the domestic domain to reduce widespread human and animal fecal contamination observed in homes, (2) pathogens detected in tubewells indicate these sources are microbiologically unsafe for drinking and were associated with child diarrhea, (3) domestic use of ponds heavily contaminated with multiple pathogens presents an under-recognized health risk, and (4) a 27 percentage point increase in improved sanitation access at village-level did not reduce detectable human fecal and pathogen contamination in this setting.

Cryptosporidium and Giardia in Humans, Domestic Animals, and Village Water Sources in Rural India.

Cryptosporidium parvum and Giardia lamblia are zoonotic enteric protozoa of significant health concern where sanitation, hygiene, and water supplies are inadequate. We examined 85 stool samples from diarrhea patients, 111 pooled fecal samples by species across seven domestic animal types, and water from tube wells (N = 207) and ponds (N = 94) across 60 villages in coastal Odisha, India, for Cryptosporidium oocysts and Giardia cysts to measure occurrence, concentration/shedding, and environmental loading rates. Oocysts/cysts were detected in 12% of diarrhea patients. Detection ranged from 0% to 35% for Cryptosporidium and 0% to 67% for Giardia across animal hosts. Animal loading estimates indicate the greatest contributors of environmental oocysts/cysts in the study region are cattle. Ponds were contaminated with both protozoa (oocysts: 37%, cysts: 74%), as were tube wells (oocysts: 10%, cysts: 14%). Future research should address the public health concern highlighted from these findings and investigate the role of domestic animals in diarrheal disease transmission in this and similar settings.

Estimating environmental conditions affecting protozoal pathogen removal in surface water wetland systems using a multi-scale, model-based approach.

Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii are waterborne protozoal pathogens distributed worldwide and empirical evidence suggests that wetlands reduce the concentrations of these pathogens under certain environmental conditions. The goal of this study was to evaluate how protozoal removal in surface water is affected by the water temperature, turbidity, salinity, and vegetation cover of wetlands in the Monterey Bay region of California. To examine how protozoal removal was affected by these environmental factors, we conducted observational experiments at three primary spatial scales: settling columns, recirculating wetland mesocosm tanks, and an experimental research wetland (Molera Wetland). Simultaneously, we developed a protozoal transport model for surface water to simulate the settling columns, the mesocosm tanks, and the Molera Wetland. With a high degree of uncertainty expected in the model predictions and field observations, we developed the model within a Bayesian statistical framework. We found protozoal removal increased when water flowed through vegetation, and with higher levels of turbidity, salinity, and temperature. Protozoal removal in surface water was maximized (~0.1 hour(-1)) when flowing through emergent vegetation at 2% cover, and with a vegetation contact time of ~30 minutes compared to the effects of temperature, salinity, and turbidity. Our studies revealed that an increase in vegetated wetland area, with water moving through vegetation, would likely improve regional water quality through the reduction of fecal protozoal pathogen loads.

Hydrologic and vegetative removal of Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii Surrogate microspheres in coastal wetlands.

Constructed wetland systems are used to reduce pollutants and pathogens in wastewater effluent, but comparatively little is known about pathogen transport through natural wetland habitats. Fecal protozoans, including Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii, are waterborne pathogens of humans and animals, which are carried by surface waters from land-based sources into coastal waters. This study evaluated key factors of coastal wetlands for the reduction of protozoal parasites in surface waters using settling column and recirculating mesocosm tank experiments. Settling column experiments evaluated the effects of salinity, temperature, and water type ("pure" versus "environmental") on the vertical settling velocities of C. parvum, G. lamblia, and T. gondii surrogates, with salinity and water type found to significantly affect settling of the parasites. The mesocosm tank experiments evaluated the effects of salinity, flow rate, and vegetation parameters on parasite and surrogate counts, with increased salinity and the presence of vegetation found to be significant factors for removal of parasites in a unidirectional transport wetland system. Overall, this study highlights the importance of water type, salinity, and vegetation parameters for pathogen transport within wetland systems, with implications for wetland management, restoration efforts, and coastal water quality.

Longitudinal Poisson regression to evaluate the epidemiology of Cryptosporidium, Giardia, and fecal indicator bacteria in coastal California wetlands.

Fecal pathogen contamination of watersheds worldwide is increasingly recognized, and natural wetlands may have an important role in mitigating fecal pathogen pollution flowing downstream. Given that waterborne protozoa, such as Cryptosporidium and Giardia, are transported within surface waters, this study evaluated associations between fecal protozoa and various wetland-specific and environmental risk factors. This study focused on three distinct coastal California wetlands: (i) a tidally influenced slough bordered by urban and agricultural areas, (ii) a seasonal wetland adjacent to a dairy, and (iii) a constructed wetland that receives agricultural runoff. Wetland type, seasonality, rainfall, and various water quality parameters were evaluated using longitudinal Poisson regression to model effects on concentrations of protozoa and indicator bacteria (Escherichia coli and total coliform). Among wetland types, the dairy wetland exhibited the highest protozoal and bacterial concentrations, and despite significant reductions in microbe concentrations, the wetland could still be seen to influence water quality in the downstream tidal wetland. Additionally, recent rainfall events were associated with higher protozoal and bacterial counts in wetland water samples across all wetland types. Notably, detection of E. coli concentrations greater than a 400 most probable number (MPN) per 100 ml was associated with higher Cryptosporidium oocyst and Giardia cyst concentrations. These findings show that natural wetlands draining agricultural and livestock operation runoff into human-utilized waterways should be considered potential sources of pathogens and that wetlands can be instrumental in reducing pathogen loads to downstream waters.