Using environmental health dialogue in a Diné-centered approach for individualized results reporting in an environmental exposure study following the Gold King Mine Spill.

BACKGROUND: On August 5, 2015, the Gold King Mine Spill (GKMS) resulted in 3 million gallons of acid mine drainage spilling into the San Juan River impacting the Diné Bikeyah (traditional homelands of the Navajo people). The Gold King Mine Spill Diné Exposure Project was formed to understand the impacts of the GKMS on the Diné (Navajo). Reporting individualized household results in an exposure study is becoming more common; however, materials are often developed with limited community input with knowledge flowing in one direction - from researcher to participant. In this study we examined the development, dissemination, and evaluation of individualized results materials. METHODS: In August 2016, Navajo Nation Community Health Representatives (Navajo CHRs) sampled household water, dust, and soil, and resident blood and urine for lead and arsenic, respectively. From May-July 2017, iterative dialogue with a wide range of community partners and a community focus groups guided the development of a culturally-based dissemination process. In August 2017, Navajo CHRs reported individualized results and they surveyed the participants on the report-back process at that time. RESULTS: All of the 63 Diné adults (100%) who participated in the exposure study received their results by a CHR in person and 42 (67%) completed an evaluation. Most of those participants (83%) were satisfied with the result packets. Respondents ranked the individual and overall household results as the most important information they received (69% and 57%, respectively), while information on metals exposures and their health effects were the least helpful. CONCLUSIONS: Our project illustrates how a model of environmental health dialogue, defined by iterative, multidirectional communication among Indigenous community members, trusted Indigenous leaders, Indigenous researchers, non-Indigenous researchers, can improve reporting individualized study results. Findings can inform future research to encourage multi-directional environmental health dialogue to craft more culturally responsive and effective dissemination and communication materials.

Sample preparation method for metal(loid) contaminant quantitation in rodent hair collected in Yuma County, Arizona.

Yuma County, Arizona, is a large agricultural hub of the USA located in the southwestern corner of Arizona on the USA-Mexico border. Year-round use of agrichemicals at a massive scale along with the influx of aquatic contaminants in the Colorado River led to significant levels of environmental pollution and hence exposure risks for people and wildlife. Although hair is a recognized biomarker for metal exposure, there is no universal hair preparation protocol. This study evaluated two digestion methods for metal quantitation using inductively coupled plasma-mass spectrometry (ICP-MS) and three methods for mercury quantitation using cold vapor-atomic absorption spectroscopy (CV-AAS), both employing certified reference materials. The "overnight" and "heating" digestion methods were suitable for ICP-MS, while only the heating method was suitable for CV-AAS. These validated methods will be useful for a variety of human and wildlife assessments of toxic metal(loid) exposure.

Traditional Ecological Knowledge Policy Considerations for Abandoned Uranium Mines on Navajo Nation.

Environmental justice is a prominent issue for Native American nations within the United States. One example is the abandoned uranium mines on the Navajo Nation that have been left unremediated since the Cold War. Often, environmental policy is developed for issues facing Native American nations that do not include input from those nations. Instead, Native American nations should have the opportunity to address environmental issues using their traditional ecological knowledge (TEK). TEK has ties to natural laws long respected by tribal communities; these laws provide the foundation for addressing the complex relationship between nature and humans. Often, policy development addressing environmental concerns is determined by non-Native American stakeholders, which can have negative effects on the Native American communities. These policies harm Native Americans rather than ultimately helping them. The focus of this discussion is how TEK can play a role in environmental policy development for the Navajo Nation surrounding abandoned uranium mines.

Uranium and arsenic unregulated water issues on Navajo lands.

The geologic profile of the western United States lends itself to naturally elevated levels of arsenic and uranium in the groundwater and can be aggravated by mining. The Navajo Nation, located in the American Southwest, is the largest contiguous Native American Nation and has over a 100-year legacy of hard rock mining. Concentrations of uranium and arsenic above drinking water standards in unregulated water sources pose various human-health risks to the Navajo Nation due to the lack of public water infrastructure that exists. Although high natural background concentrations may occur in some environments, anthropogenic contamination concerns are especially troublesome for the Navajo Nation, where past uranium mining activity and natural sources affect unregulated water supplies. Community engaged research on uranium and arsenic present in unregulated water wells in the western portion of the Navajo Nation has been a focus of the Ingram laboratory since 2003. These studies have provided important information, particularly for uranium and arsenic, to the communities and the Navajo tribal leaders.

Advancing Cancer Workforce Capacity for American Indians and Alaska Natives: The Development of a Validated System to Optimize Trainee Participation and Outcome Tracking.

Although American Indian and Alaska Native (AIAN) students are the most underrepresented group in the U.S. in biomedical and health sciences relative to population size, little is known about long-term research education programs and outcome tracking. For over 20 years, the Partnership for Native American Cancer Prevention (NACP) has been supported under the National Cancer Institute's (NCI)-funded Comprehensive Partnerships to Advance Cancer Health Equity (CPACHE) program. Programming included hands-on mentored research and an array of development opportunities. A validated tracking system combining participation records, institutional records, and enrollment/degree attainment from the National Student Clearinghouse documents outcomes. Collectively (2002-2022) NACP engaged 367 AIAN trainees, of whom 237 individuals earned 220 bachelors, 87 masters, and 34 doctoral/professional degrees. Approximately 45% of AIAN doctoral recipients are currently engaged in academic or clinical work, and 10% in industry or tribal leadership. A total of 238 AIAN students participated in mentored research, with 85% demonstrating strong outcomes; 51% attained a degree, and 34% are currently enrolled. Implementation of a robust tracking system documented acceleration in degree attainment over time. Next steps will evaluate the most impactful training activities on student outcomes.

Perspective Developing Successful Collaborative Research Partnerships with AI/AN Communities.

In the United States, American Indian and Alaska Native (AI/AN) people are frequently under- or misrepresented in research and health statistics. A principal reason for this disparity is the lack of collaborative partnerships between researchers and tribes. There are hesitations from both academic Western scientists and tribal communities to establish new partnerships due to differences in cultural and scientific understanding, from data ownership and privacy to dissemination and project expansion. An infamous example is the mishandling of samples collected from the Havasupai Tribe by Arizona State University (ASU) scientists, leading to a legal battle between the tribe and ASU and ending in a moratorium of research with the Havasupai people. This paper will explore three successful and positive collaborations with a large and small tribe, including how the partnerships were established and the outcomes of the collaboration. In addition, the paper will provide perspective of what needs to be addressed by Western scientists if productive collaborations with tribal groups are to be established.

Approaches for Disseminating Environmental Research Findings to Navajo Communities.

We report the use of three different dissemination approaches for providing environmental research results back to Navajo communities from different research projects. The objectives of the dissemination are to provide the results to the community, have a dialogue about the results, and learn more about the environmental concerns of the community for potential future research projects. The first approach utilizes radio announcements and flyers provided to the community announcing dissemination meetings specific to the research projects. The second approach is more collaborative, working with a grassroots organization to organize report-back meetings, as well as one-on-one discussions of the research project. The third approach involves the development of a booklet for distribution to communities along with an oral presentation at the regularly scheduled monthly community meetings to discuss the information. Overall, the second and third approaches are more effective than the first approach in terms of dissemination to a larger number of community members, as well as increased dialogue between the researchers and the communities.

NACP: Partnership for Native American Cancer Prevention.

Cancer trends over a two-decade period show a greater reduction in cancer mortality rates for non-Hispanic Whites than for Native Americans. The Partnership for Native American Cancer Prevention (NACP) was established to address cancer health disparities that impact Native Americans. The partners are Northern Arizona University, the University of Arizona Cancer Center, Arizona's tribal communities and the National Cancer Institute. The activities include outreach, research and cancer education. Overall, NACP seeks to expand capacity for culturally-sensitive and community-relevant research on cancer, and to continue developing respectful collaborations that will empower sovereign Native American communities to define, implement, and achieve their goals for cancer health equity.

Dissolved Uranium and Arsenic in Unregulated Groundwater Sources - Western Navajo Nation.

Concentrations of dissolved uranium (U) and arsenic (As) above drinking water standards in unregulated water sources pose various human health risks. Although high natural background concentrations may occur in some environments (Runnells et al. 1992), anthropogenic contamination concerns are especially troublesome on the Navajo Nation (NN), where past U mining activity may have contaminated water supplies. This research investigated U and As groundwater contamination issues in unregulated wells in the western portion of the NN. Objectives of this research were to provide insights to human health risks by assessing the spatial extent and seasonal variability of U and As concentrations while effectively communicating the potential contamination risks to the local Navajo people. Eighty-two unregulated wells were sampled in 2018; nine of these sources exceeded the maximum contaminant level (MCL) for drinking water standards for U (30 µg/L), and 14 exceeded the MCL for drinking water standards for As (10 µg/L). U and As levels were highest in the southwest portion of the study area and seasonal variability was observed in a subset of wells, especially shallower hand dug wells and hand pumps. The results were compiled into a report that was presented to NN chapters included in the study as well as the Navajo Department of Water Resources and the NN Environmental Protection Agency. Implications for regional water quality patterns can help to direct policy recommendations for well monitoring, water use, and remediation targets.

Quantification of Elemental Contaminants in Unregulated Water across Western Navajo Nation.

The geologic profile of the western United States lends itself to naturally elevated levels of arsenic and uranium in groundwater and can be exacerbated by mining enterprises. The Navajo Nation, located in the American Southwest, is the largest contiguous Native American Nation and has over a 100-year legacy of hard rock mining. This study has two objectives, quantify the arsenic and uranium concentrations in water systems in the Arizona and Utah side of the Navajo Nation compared to the New Mexico side and to determine if there are other elements of concern. Between 2014 and 2017, 294 water samples were collected across the Arizona and Utah side of the Navajo Nation and analyzed for 21 elements. Of these, 14 elements had at least one instance of a concentration greater than a national regulatory limit, and six of these (V, Ca, As, Mn, Li, and U) had the highest incidence of exceedances and were of concern to various communities on the Navajo Nation. Our findings are similar to other studies conducted in Arizona and on the Navajo Nation and demonstrate that other elements may be a concern for public health beyond arsenic and uranium.

Traditional Sheep Consumption by Navajo People in Cameron, Arizona.

Over 500 abandoned uranium mines are located on the Navajo Reservation. Different pathways of environmental uranium exposure have been studied with respect to the Navajo people including water, soil, and plants; however, uranium exposure from traditional Navajo food, specifically mutton (sheep), has not been reported. This study focuses on mutton consumption in the small community of Cameron, Arizona, located in the southwestern region of the Navajo Nation and initiated after community members expressed concern with the uranium exposure of their sheep. Preliminary investigation into the presence of uranium in sheep raised near Cameron showed elevated uranium levels in the kidneys the sheep tested. The goal of this study is to investigate mutton consumption among the Navajo living in Cameron. Mutton is a traditional food of the Navajo, but consumption practices are not well documented. An important aspect of determining the extent of exposure through food consumption is to assess the frequency of consumption. The results of this study indicate the Cameron participants consume mutton most commonly at family gatherings or celebrations. The survey suggests that less mutton is consumed now compared to the past, and there is concern that contaminated mutton may change traditional ceremonies.

Metal Transformation by a Novel Pelosinus Isolate From a Subsurface Environment.

The capability of microorganisms to alter metal speciation offers potential for the development of new strategies for immobilization of toxic metals in the environment. A metal-reducing microbe, "Pelosinus lilae" strain UFO1, was isolated under strictly anaerobic conditions from an Fe(III)-reducing enrichment established with uncontaminated soil from the Department of Energy Oak Ridge Field Research Center, Tennessee. "P. lilae" UFO1 is a rod-shaped, spore-forming, and Gram-variable anaerobe with a fermentative metabolism. It is capable of reducing the humic acid analog anthraquinone-2,6-disulfonate (AQDS) using a variety of fermentable substrates and H2. Reduction of Fe(III)-nitrilotriacetic acid occurred in the presence of lactate as carbon and electron donor. Ferrihydrite was not reduced in the absence of AQDS. Nearly complete reduction of 1, 3, and 5 ppm Cr(VI) occurred within 24 h in suspensions containing 108 cells mL-1 when provided with 10 mM lactate; when 1 mM AQDS was added, 3 and 5 ppm Cr(VI) were reduced to 0.1 ppm within 2 h. Strain UFO1 is a novel species within the bacterial genus Pelosinus, having 98.16% 16S rRNA gene sequence similarity with the most closely related described species, Pelosinus fermentans R7T. The G+C content of the genomic DNA was 38 mol%, and DNA-DNA hybridization of "P. lilae" UFO1 against P. fermentans R7T indicated an average 16.8% DNA-DNA similarity. The unique phylogenetic, physiologic, and metal-transforming characteristics of "P. lilae" UFO1 reveal it is a novel isolate of the described genus Pelosinus.

Spatial distribution of uranium in mice kidneys detected by laser ablation inductively coupled plasma mass spectrometry.

The aim of the study is to better understand where uranium deposits in mice kidneys. The spatial distribution of uranium was examined in the kidneys of C57BL/6 mice using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Mice were exposed to varying levels of uranyl nitrate in their drinking water. Calibration standards were developed to allow for semi-quantitative measurement of uranium in the cortical and medullary regions of mice kidney by LA-ICP-MS. Scanning electron microscopy was used to image the ablation patterns on the kidney. Uranium levels were observed to increase in kidney tissue as uranyl nitrate treatment exposure levels increased. A trend towards a higher uranium concentration in the medullary versus cortical region of the kidneys was observed. These results show the usefulness of LA-ICP-MS in toxicity studies by providing a quantitative, spatial assessment of uranium deposition in a target organ.

Health Effects and Environmental Justice Concerns of Exposure to Uranium in Drinking Water.

We discuss the recent epidemiologic literature regarding health effects of uranium exposure in drinking water focusing on the chemical characteristics of uranium. While there is strong toxicologic evidence for renal and reproductive effects as well as DNA damage, the epidemiologic evidence for these effects in people exposed to uranium in drinking water is limited. Further, epidemiologic evidence is lacking for cardiovascular and oncogenic effects. One challenge in characterizing health effects of uranium in drinking water is the paucity of long-term cohort studies with individual level exposure assessment. Nevertheless, there are environmental justice concerns due to the substantial exposures for certain populations. For example, we present original data suggesting that individuals living in the Navajo Nation are exposed to high levels of uranium in unregulated well water used for drinking. In 10 out of 185 samples (5.4 %), concentrations of uranium exceeded standards under the Safe Drinking Water Act. Therefore, efforts to mitigate exposure to toxic elements in drinking water are warranted and should be prioritized.

Detection of fatty acids from intact microorganisms by molecular beam static secondary ion mass spectrometry.

We report the use of a surface analysis approach, static secondary ion mass spectrometry (SIMS) equipped with a molecular (ReO(4)(-)) ion primary beam, to analyze the surface of intact microbial cells. SIMS spectra of 28 microorganisms were compared to fatty acid profiles determined by gas chromatographic analysis of transesterfied fatty acids extracted from the same organisms. The results indicate that surface bombardment using the molecular primary beam cleaved the ester linkage characteristic of bacteria at the glycerophosphate backbone of the phospholipid components of the cell membrane. This cleavage enables direct detection of the fatty acid conjugate base of intact microorganisms by static SIMS. The limit of detection for this approach is approximately 10(7) bacterial cells/cm(2). Multivariate statistical methods were applied in a graded approach to the SIMS microbial data. The results showed that the full data set could initially be statistically grouped based upon major differences in biochemical composition of the cell wall. The gram-positive bacteria were further statistically analyzed, followed by final analysis of a specific bacterial genus that was successfully grouped by species. Additionally, the use of SIMS to detect microbes on mineral surfaces is demonstrated by an analysis of Shewanella oneidensis on crushed hematite. The results of this study provide evidence for the potential of static SIMS to rapidly detect bacterial species based on ion fragments originating from cell membrane lipids directly from sample surfaces.

Preservation of yeast cell morphology for scanning electron microscopy using 3.28-micro m IR laser irradiation.

As an alternative to conventional fixation procedures for scanning electron microscopy (SEM) analysis, yeast cells (Saccharomyces cerevisiae) were irradiated in ambient air, with an intense 3.28-micro m IR laser pulse. The morphology of the irradiated cells was well preserved, while nonirradiated control cells were severely shriveled.

Uranium Leaching from Contaminated Soil Utilizing Rhamnolipid, EDTA, and Citric Acid.

Biosurfactants have recently gained attention as "green" agents that can be used to enhance the remediation of heavy metals and some organic matter in contaminated soils. The overall objective of this paper was to investigate rhamnolipid, a microbial produced biosurfactant, and its ability to leach uranium present in contaminated soil from an abandoned mine site. Soil samples were collected from two locations in northern Arizona: Cameron (site of open pit mining) and Leupp (control-no mining). The approach taken was to first determine the total uranium content in each soil using a hydrofluoric acid digestion, then comparing the amount of metal removed by rhamnolipid to other chelating agents EDTA and citric acid, and finally determining the amount of soluble metal in the soil matrix using a sequential extraction. Results suggested a complex system for metal removal from soil utilizing rhamnolipid. It was determined that rhamnolipid at a concentration of 150 µM was as effective as EDTA but not as effective as citric acid for the removal of soluble uranium. However, the rhamnolipid was only slightly better at removing uranium from the mining soil compared to a purified water control. Overall, this study demonstrated that rhamnolipid ability to remove uranium from contaminated soil is comparable to EDTA and to a lesser extent citric acid, but, for the soils investigated, it is not significantly better than a simple water wash.

Micropatterned avidin arrays on silicon substrates via photolithography, self-assembly and bioconjugation.

Processes for micropatterning protein arrays on inorganic substrates have gained attention in the development of biosensors and clinical diagnostics. This study demonstrates a chemically selective method based on photolithographic deposition of gold patterns with the subsequent attachment of functionalized alkanethiols via molecular self-assembly. Selective capping of carboxy groups on alkanethiols by N-hydroxysuccinimide esters allowed the deposition of methoxypoly(ethylene glycol) silane as a blocking agent on unpatterned regions. Carboxylates were used to couple a form of avidin to create a microarray of protein. This microarray was successfully probed with biotinylated quantum dots. In-process characterization methods included grazing-angle Fourier-transform IR spectroscopy, ellipsometry, contact-angle goniometry, atomic-force microscopy and fluorescence microscopy.