Childhood parasitic infections and gastrointestinal illness in indigenous communities at Lake Atitlán, Guatemala.

Lake Atitlán has experienced a decline in water quality resulting from cultural eutrophication. Indigenous Mayans who already face disproportionate health challenges rely directly on the lake water. Our objectives were to: (1) estimate prevalence of shedding of water-borne fecal parasites among children 5 years of age and younger, (2) assess household-reported incidence of gastrointestinal illness in children, and (3) characterize water sources, treatment, and sanitation conditions in households. We hypothesized that household use of untreated lake water results in increased risk of shedding of parasites and gastrointestinal symptoms. A cross-sectional fecal sampling and physical exam of 401 children were conducted along with WASH surveys in partnership with healthcare providers in seven communities. Fecal samples were screened for Giardia lamblia and Cryptosporidium parvum, using a rapid ELISA, with a portion examined by microscope. The prevalence of parasite shedding was 12.2% (9.7% for Giardia; 2.5% for Cryptosporidium). Risk factors for Giardia shedding included age 3 years or older (3.4 odds ratio, z-stat = 2.781 p = 0.0054), low height-for-age z-score (2.3 odds ratio, z-stat = 2.225, p = 0.0216), lack of any household water treatment (2.5 odds ratio, z-stat = 2.492, p < 0.0012), and open access to household latrine (2.04 odds ratio, z-stat = 1.992, p = 0.0464). The majority (77.3%) of households reported water treatment, boiling and gravity fed filters as the most widespread practices. The vast majority of households (92%) reported usage of a latrine, while 40% reported open and shared access beyond their household. An overwhelming majority of households reported diarrhea and fever several times per year or greater, with approximately half reporting vomiting at that frequency. Lake water use was identified as a risk factor for households reporting frequent gastrointestinal symptoms (odds ratio of 2.5, 4.4, and 2.6; z-stat of 3.10, 3.65, and 3.0; p-values of 0.0021, 0.0003, and 0.0028, for diarrhea, vomiting, and fever, respectively) in children 5 years of age and younger. The frequency of gastrointestinal illness with a strong link to lake drinking water cannot be explained by the prevalence of protozoa, and risk from other enteropathogens must be explored. Improving access to water treatment and sanitation practices could substantially reduce the parasite burden faced by developing children in the region.

Harmful Algal Blooms Threaten the Health of Peri-Urban Fisher Communities: A case study in Kisumu Bay, Lake Victoria, Kenya.

Available guidance to mitigate health risks from exposure to freshwater harmful algal blooms (HABs) is largely derived from temperate ecosystems. Yet in tropical ecosystems, HABs can occur year-round, and resource-dependent populations face multiple routes of exposure to toxic components. Along Winam Gulf, Lake Victoria, Kenya, fisher communities rely on lake water contaminated with microcystins (MCs) from HABs. In these peri-urban communities near Kisumu, we tested hypotheses that MCs exceed exposure guidelines across seasons, and persistent HABs present a chronic risk to fisher communities through ingestion with minimal water treatment and frequent, direct contact. We tested source waters at eleven communities across dry and rainy seasons from September 2015 through May 2016. We measured MCs, other metabolites, physicochemical parameters, chlorophyll a, phytoplankton abundance and diversity, and fecal indicators. We then selected four communities for interviews about water sources, usage, and treatment. Greater than 30% of source water samples exceeded WHO drinking water guidelines for MCs (1µg/L), and over 60% of source water samples exceeded USEPA guidelines for children and immunocompromised individuals. 50% of households reported sole use of raw lake water for drinking and household use, with alternate sources including rain and boreholes. Household chlorination was the most widespread treatment utilized. At this tropical, eutrophic lake, HABs pose a year-round health risk for fisher communities in resource -limited settings. Community-based solutions and site-specific guidance for Kisumu Bay and similarly impacted regions is needed to address a chronic health exposure likely to increase in severity and duration with global climate change.

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.

Combined Danio rerio embryo morbidity, mortality and photomotor response assay: A tool for developmental risk assessment from chronic cyanoHAB exposure.

Freshwater harmful algal blooms produce a broad array of bioactive compounds, with variable polarity. Acute exposure to cyanotoxins can impact the liver, nervous system, gastrointestinal tract, skin, and immune function. Increasing evidence suggests chronic effects from low-level exposures of cyanotoxins and other associated bioactive metabolites of cyanobacterial origin. These sundry compounds persist in drinking and recreational waters and challenge resource managers in detection and removal. A systematic approach to assess the developmental toxicity of cyanobacterial metabolite standards was employed utilizing a robust and high throughput developmental Danio rerio embryo platform that incorporated a neurobehavioral endpoint, photomotor response. Subsequently, we applied the platform to cyanobacterial bloom surface water samples taken from temperate recreational beaches and tropical lake subsistence drinking water sources as a model approach. Dechorionated Danio rerio embryos were statically immersed beginning at four to six hours post fertilization at environmentally relevant concentrations, and then assessed at 24 h and 5 days for morbidity, morphological changes, and photomotor response. At least one assessed endpoint deviated significantly for exposed embryos for 22 out of 25 metabolites examined. Notably, the alkaloid lyngbyatoxin-a resulted in profound, dose-dependent morbidity and mortality beginning at 5 µg/L. In addition, hydrophobic components of extracts from beach monitoring resulted in potent morbidity and mortality despite only trace cyanotoxins detected. The hydrophilic extracts with several order of magnitude higher concentrations of microcystins resulted in no morbidity or mortality. Developmental photomotor response was consistently altered in environmental bloom samples, independent of the presence or concentration of toxins detected in extracts. While limited with respect to more polar compounds, this novel screening approach complements specific fingerprinting of acutely toxic metabolites with robust assessment of developmental toxicity, critical for chronic exposure scenarios.

Employing CBPR to investigate function, utility, and longevity of household filters to improve potable water quality for indigenous peoples at Lake Atitlán, Guatemala: a pilot study with San Pedro de La Laguna.

Cyanobacterial blooms at Lake Atitlán in Guatemala threaten and compromise the livelihood and health of local residents. Indigenous Tz'utujil, Kaqchikel, and K'iche' rely directly on lake water for drinking, bathing, cleaning, cooking, and fishing. Nonpoint source runoff and untreated wastewater pumped directly into the lake contribute to high fecal pathogen loads into source waters. Concurrent nutrient loading results in cyanobacterial blooms further compromising water quality. A lakeside municipality facing high rates of childhood gastrointestinal illness volunteered to engage in community-based participatory research (CBPR) to evaluate efficacy, utility, and longevity of filters in households. The filters consistently reduced the risk of coliforms and E. coli in household water drawn from the lake based on World Health Organization guidelines. Household surveys were simultaneously administered through a student leadership group regarding water usage, water quality, and community health. Filters demonstrated ability to reduce high loads of fecal indicators from source waters and ability to remove a cyanobacterial toxin (microcystin) at 10 µg/L in deionized water. Further studies are imperative to determine longevity of use in households and CBPR provides a powerful avenue to test efficacy of a possible intervention while engaging stakeholders and empowering community members with sustainable solutions.

Composition and design of vegetative filter strips instrumental in improving water quality by mass reduction of suspended sediment, nutrients and Escherichia coli in overland flows in eastern escarpment of Mau Forest, Njoro River Watershed, Kenya.

This study assessed the effect of vegetative filter strip (VFS) in removal of suspended sediment (SS), total nitrogen, total phosphorus and Escherichia coli (E. coli) in overland flow to improve receiving water quality standards. Four and half kilograms of cowpat manure was applied to the model pasture 14 m beyond the edge of vegetated filter strip (VFS) comprising 10-m Napier grass draining into 20-m Kikuyu grass (VFS II), 10-m Kikuyu grass draining into 20-m Napier grass (VFS III) and native grass mixture of Couch-Buffel (VFS I-control). Overland flow water samples were collected from the sites at positions 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 and 30 m along the length of VFSs. E. coli removal by Napier grass VFS was on the order of log unit, which provided an important level of protection and reduced surface-flow concentrations of E. coli to below the 200 (CFU 100 mL-1) recommended water quality standards, but not for nutrients and SS. The Napier grass showed highest efficiency (99.6 %), thus outperforming both Kikuyu grass (85.8 %) and Couch-Buffel grasses VFS (67.9 ± 4.2 %) in removing E. coli from overland flow. The low-level efficiency of native Couch-Buffel grasses in reducing E. coli in overland flow was because of preferential flow. Composition and design of VFS was instrumental and could be applied with a high potential of contracting the uncertainty in improving water quality standards through mass reduction of SS, nutrients and E. coli load in watersheds.

Aggregate culture: A more accurate predictor of microcystin toxicity for risk assessment.

Aggregate or spheroid culture has emerged as a more biologically relevant method for screening pharmaceutical compounds and understanding exact mechanism of action. Here in, the aggregate approach applied to the freshwater toxins, microcystins, further unearths exact mechanism(s) of toxicity and provides a markedly improved in vitro predictor of toxicity. Microcystins result in acute intoxication by binding covalently to protein phosphatase 1/2A, resulting in hepatocellular necrosis, hemorrhaging and death. Hepatocellular uptake by organic anion transporting polypeptides (OATPs), in addition to other intracellular sequelae, is considered essential for toxicity. In aggregate HepG2, expression of OAT1B1 and OATP1B3 significantly increased relative to monolayer culture. Uptake of two fluorescently labeled substrates significantly increased in aggregates compared with monolayer, confirmed by inhibition of uptake with known competitive substrates. Increased reaction oxygen species (ROS) production occurred following a three-hour exposure of microcystin LR at concentrations from 100 nM to 100 µM, with reversal by ROS scavengers, in contrast with no response in monolayers. These results suggest monolayer culture inadequately predict intracellular effects of microcystins and support evidence that aggregate culture more closely approximates in vivo form and function. The approach results in more reliable prediction of microcystin toxicity in vitro.

Rapid quantitative analysis of microcystins in raw surface waters with MALDI MS utilizing easily synthesized internal standards.

The freshwater cyanotoxins, microcystins (MCs), pose a global public health threat as potent hepatotoxins in cyanobacterial blooms; their persistence in drinking and recreational water has been associated with potential chronic effects in addition to acute intoxications. Rapid and accurate detection of the over 80 structural congeners is challenged by the rigorous and time consuming clean up required to overcome interference found in raw water samples. MALDI-MS has shown promise for rapid quantification of individual congeners in raw water samples, with very low operative cost, but so far limited sensitivity and lack of available and versatile internal standards (ISs) has limited its use. Two easily synthesized S-hydroxyethyl-Cys(7)-MC-LR and -RR ISs were used to generate linear standard curves in a reflectron MALDI instrument, reproducible across several orders of magnitude for MC-LR, -RR and -YR. Minimum quantification limits in direct water samples with no clean up or concentration step involved were consistently below 7 µg/L, with recoveries from spiked samples between 80 and 119%. This method improves sensitivity by 30 fold over previous reports of quantitative MALDI-TOF applications to MCs and provides a salient option for rapid throughput analysis for multiple MC congeners in untreated raw surface water blooms as a means to identify source public health threats and target intervention strategies within a watershed. As demonstrated by analysis of a set of samples from Uruguay, utilizing the reaction of different MC congeners with alternate sulfhydryl compounds, the m/z of the IS can be customized to avoid overlap with interfering compounds in local surface water samples.

Microcystins in potable surface waters: toxic effects and removal strategies.

In freshwater, harmful cyanobacterial blooms threaten to increase with global climate change and eutrophication of surface waters. In addition to the burden and necessity of removal of algal material during water treatment processes, bloom-forming cyanobacteria can produce a class of remarkably stable toxins, microcystins, difficult to remove from drinking water sources. A number of animal intoxications over the past 20 years have served as sentinels for widespread risk presented by microcystins. Cyanobacterial blooms have the potential to threaten severely both public health and the regional economy of affected communities, particularly those with limited infrastructure or resources. Our main objectives were to assess whether existing water treatment infrastructure provides sufficient protection against microcystin exposure, identify available options feasible to implement in resource-limited communities in bloom scenarios and to identify strategies for improved solutions. Finally, interventions at the watershed level aimed at bloom prevention and risk reduction for entry into potable water sources were outlined. We evaluated primary studies, reviews and reports for treatment options for microcystins in surface waters, potable water sources and treatment plants. Because of the difficulty of removal of microcystins, prevention is ideal; once in the public water supply, the coarse removal of cyanobacterial cells combined with secondary carbon filtration of dissolved toxins currently provides the greatest potential for protection of public health. Options for point of use filtration must be optimized to provide affordable and adequate protection for affected communities.

Public health implications of lead poisoning in backyard chickens and cattle: four cases.

Lead intoxication in livestock has historically been associated with cattle turned out to pasture and accidental ingestion of lead from drinking crankcase oil, licking grease from machinery, chewing on plumbing or batteries, or drinking water contaminated from leaching materials. Even with the decrease in manufactured items produced with lead, contaminants persist in the landscape and may enter the food supply through animal products. Changing patterns of open range herds moving to new pasture and the increased popularity of urban/suburban backyard chickens or other livestock necessitates public awareness about the clinical signs of lead intoxication, the potential for subclinical animals, public health concerns, particularly for exposure in children, and testing options available. Cases of lead intoxication in livestock demand a thorough case work-up to identify all sources of lead, address subclinical cases, evaluate risk to consumers, and make management suggestions for future prevention. We discuss four recent cases of confirmed lead poisoning in backyard chickens and open range cattle and assess the public health implications therein. Taken as a whole and considering the potential of the remaining herd or flock to be affected without necessarily showing signs, public health officials and veterinarians should be prepared to advise clients on case work-up and management and prevention considerations. Backyard chickens and cattle may not present for suspected lead poisoning as in several of the cases discussed herein yet may still contain concerning tissue or blood levels. The authors believe increased surveillance through heavy metal screens is crucial to adequately protect public health.