Range-wide genetic analysis of an endangered bumble bee (Bombus affinis, Hymenoptera: Apidae) reveals population structure, isolation by distance, and low colony abundance.

Declines in bumble bee species range and abundances are documented across multiple continents and have prompted the need for research to aid species recovery and conservation. The rusty patched bumble bee (Bombus affinis) is the first federally listed bumble bee species in North America. We conducted a range-wide population genetics study of B. affinis from across all extant conservation units to inform conservation efforts. To understand the species' vulnerability and help establish recovery targets, we examined population structure, patterns of genetic diversity, and population differentiation. Additionally, we conducted a site-level analysis of colony abundance to inform prioritizing areas for conservation, translocation, and other recovery actions. We find substantial evidence of population structuring along an east-to-west gradient. Putative populations show evidence of isolation by distance, high inbreeding coefficients, and a range-wide male diploidy rate of ~15%. Our results suggest the Appalachians represent a genetically distinct cluster with high levels of private alleles and substantial differentiation from the rest of the extant range. Site-level analyses suggest low colony abundance estimates for B. affinis compared to similar datasets of stable, co-occurring species. These results lend genetic support to trends from observational studies, suggesting that B. affinis has undergone a recent decline and exhibit substantial spatial structure. The low colony abundances observed here suggest caution in overinterpreting the stability of populations even where B. affinis is reliably detected interannually. These results help delineate informed management units, provide context for the potential risks of translocation programs, and help set clear recovery targets for this and other threatened bumble bee species.

Screening for health literacy, social determinants, and discrimination in health plans.

OBJECTIVES: Health inequities are frequently driven by social determinants of health (SDOH) and structural determinants of health. Our pilot sought to test the feasibility of screening for health literacy (HL) and perceived health care discrimination (PHD) through a live telephonic-facilitated survey experience with managed care patients. STUDY DESIGN: Cross-sectional study. METHODS: Newly enrolled Medicare Advantage patients were screened for self-reported PHD, HL, and multiple SDOH using validated screening tools. Response rates for both HL and PHD screens were analyzed. A χ2 test for association between response to PHD screen and patient race was conducted. A weighted logistic regression model was used to understand how HL is associated with SDOH and demographic factors (age, gender, race/ethnicity, and income). RESULTS: HL and PHD screening questions have different levels of feasibility. Administering the HL screen did not present a challenge, and patients felt comfortable responding to it. On the other hand, the PHD question had a lower response rate among patients, and some concierge advocates felt uncomfortable asking patients the question. Based on the self-reported HL data collected, low/limited HL is associated with patients who were Black, were low income, reported loneliness or isolation, or reported food insecurity. It is important to note that the study's findings are limited by the small sample size and that study results do not imply causality. CONCLUSIONS: It is feasible to collect self-reported HL data through a live telephonic format at the time of patient enrollment into a health plan. Health plans can leverage such screenings to better understand patient barriers for health equity-oriented interventions.

Engineering Vacancies for the Creation of Antisite Defects in Chemical Vapor Deposition Grown Monolayer MoS2 and WS2 via Proton Irradiation.

It is critical to understand the laws of quantum mechanics in transformative technologies for computation and quantum information science applications to enable the ongoing second quantum revolution calls. Recently, spin qubits based on point defects have gained great attention, since these qubits can be initiated, selectively controlled, and read out with high precision at ambient temperature. The major challenge in these systems is controllably generating multiqubit systems while properly coupling the defects. To address this issue, we began by tackling the engineering challenges these systems present and understanding the fundamentals of defects. In this regard, we controllably generate defects in MoS2 and WS2 monolayers and tune their physicochemical properties via proton irradiation. We quantitatively discovered that the proton energy could modulate the defects' density and nature; higher defect densities were seen with lower proton irradiation energies. Three distinct defect types were observed: vacancies, antisites, and adatoms. In particular, the creation and manipulation of antisite defects provides an alternative way to create and pattern spin qubits based on point defects. Our results demonstrate that altering the particle irradiation energy can regulate the formation of defects, which can be utilized to modify the properties of 2D materials and create reliable electronic devices.

Using the stress-vulnerability model to better understand suicide in prison populations.

Suicide is the culmination of the interaction of a number of factors with the critical component being distress. The contribution of mental illness as a vulnerability factor in an individual case of suicide by a prisoner may be significant, marginal or non-existent. Because of the high prevalence in prison populations of all risk factors for suicide, relying solely on those factors is of little use in elaborating the risk of suicide for a given prisoner. Whilst a current mental illness or past history of a suicide attempt are two factors that appear to be strongly associated with completed suicide, no screening or risk assessment tool has any proven efficacy in predicting suicide in prison populations. The stress-vulnerability model offers a more sophisticated and comprehensive assessment and also informs a more individualised needs-based management of the risk of suicide with an emphasis on detecting and responding to a prisoner's distress.

Chromosome-scale genome assembly of the rusty patched bumble bee, Bombus affinis (Cresson) (Hymenoptera: Apidae), an endangered North American pollinator.

The rusty patched bumble bee, Bombus affinis, is an important pollinator in North America and a federally listed endangered species. Due to habitat loss and large declines in population size, B. affinis is facing imminent extinction unless human intervention and recovery efforts are implemented. To better understand B. affinis biology and population genetic and genomic landscapes, we sequenced and assembled the B. affinis genome from a single haploid male. Whole genome HiFi sequencing on PacBio coupled with HiC sequencing resulted in a complete and highly contiguous contig assembly that was scaffolded into a chromosomal context, resolving 18 chromosomes distributed across the 365.1 Mb assembly. All material for both HiFi and HiC sequencing was derived from a single abdominal tissue segment from the single male. These assembly results, coupled with the minimal amount of tissue destructively sampled, demonstrate methods for generating contiguous and complete genomic resources for a rare and endangered species with limited material available and highlight the importance of sample preservation. Precise methods and applications of these methods are presented for potential applications in other species with similar limitations in specimen availability and curation considerations.

Addressing Detection Uncertainty in Bombus affinis (Hymenoptera: Apidae) Surveys Can Improve Inferences Made From Monitoring.

The U.S. Fish and Wildlife Service developed national guidelines to track species recovery of the endangered rusty patched bumble bee [Bombus affinis Cresson (Hymenoptera: Apidae)] and to investigate changes in species occupancy across space and time. As with other native bee monitoring efforts, managers have specifically acknowledged the need to address species detection uncertainty and determine the sampling effort required to infer species absence within sites. We used single-season, single-species occupancy models fit to field data collected in four states to estimate imperfect detection of B. affinis and to determine the survey effort required to achieve high confidence of species detection. Our analysis revealed a precipitous, seasonal, decline in B. affinis detection probability throughout the July through September sampling window in 2021. We estimated that six, 30-min surveys conducted in early July are required to achieve a 95% cumulative detection probability, whereas >10 surveys would be required in early August to achieve the same level of confidence. Our analysis also showed B. affinis was less likely to be detected during hot and humid days and at patches of reduced habitat quality. Bombus affinis was frequently observed on Monarda fistulosa (Lamiales: Lamiaceae), followed by [Pycnanthemum virginianum Rob. and Fernald (Lamiales: Lamiaceae)], Eutrochium maculatum Lamont (Asterales: Asteraceae), and Veronicastrum virginicum Farw. (Lamiales: Plantaginaceae). Although our research is focused on B. affinis, it is relevant for monitoring other bumble bees of conservation concern, such as B. occidentalis Greene (Hymenoptera: Apidae) and B. terricola Kirby (Hymenoptera: Apidae) for which monitoring efforts have been recently initiated and occupancy is a variable of conservation interest.

A fully automatic framework for evaluating cosmetic results of breast conserving therapy.

The breast cosmetic outcome after breast conserving therapy is essential for evaluating breast treatment and determining patient's remedy selection. This prompts the need of objective and efficient methods for breast cosmesis evaluations. However, current evaluation methods rely on ratings from a small group of physicians or semi-automated pipelines, making the processes time-consuming and their results inconsistent. To solve the problem, in this study, we proposed: 1. a fully-automatic Machine Learning Breast Cosmetic evaluation algorithm leveraging the state-of-the-art Deep Learning algorithms for breast detection and contour annotation, 2. a novel set of Breast Cosmesis features, 3. a new Breast Cosmetic dataset consisting 3k+ images from three clinical trials with human annotations on both breast components and their cosmesis scores. We show our fully-automatic framework can achieve comparable performance to state-of-the-art without the need of human inputs, leading to a more objective, low-cost and scalable solution for breast cosmetic evaluation in breast cancer treatment.

Incorporation of Ti in epitaxial Fe2TiO4thin films.

The titanomagnetites (Fe2-xTixO4,x⩽ 1) are a family of reducible spinel-structure oxides of interest for their favorable magnetic, catalytic, and electrical transport properties. To understand the stability of the system during low temperature deposition, epitaxial thin films of Fe2TiO4were deposited by molecular beam epitaxy (MBE) on MgO(001) at 250-375 °C. The homogeneous incorporation of Ti, Fe valence state, and film morphology were all found to be strongly dependent on the oxidation conditions at the low substrate temperatures employed. More oxidizing conditions led to phase separation into epitaxial, faceted Fe3O4and rutile TiO2. Less oxidizing conditions resulted in polycrystalline films that exhibited Ti segregation to the film surface, as well as mixed Fe valence (Fe3+, Fe2+, Fe0). A narrow window of intermediate oxygen partial pressure during deposition yielded nearly homogeneous Ti incorporation and a large fraction of Fe2+. However, these films were poorly crystallized, and no occupation of tetrahedral sites in the spinel lattice by Fe2+was detected by x-ray magnetic circular dichroism at the Fe L-edge. After vacuum annealing, a small fraction of Fe2+was found to occupy tetrahedral sites. Comparison of these results with previous work suggests that the low temperature deposition conditions imposed by use of MgO substrates limits the incorporation of Ti into the spinel lattice. This work suggests a path towards obtaining stoichiometric, well-crystallized Fe2TiO4by MBE by utilizing high substrate temperature and low oxygen partial pressure during deposition on thermally stable substrates.

Electronic and structural properties of single-crystal Jahn-Teller active Co1+x Mn2-x O4 thin films.

Recent investigations on spinel CoMn2O4 have shown its potential for applications in water splitting and fuel cell technologies as it exhibits strong catalytic behavior through oxygen reduction reactivity. To further understand this material, we report for the first time the synthesis of single-crystalline Co1+x Mn2-x O4 thin films using molecular beam epitaxy. By varying sample composition, we establish links between cation stoichiometry and material properties using in-situ x-ray photoelectron spectroscopy, x-ray diffraction, scanning transmission electron microscopy, x-ray absorption spectroscopy, and spectroscopic ellipsometry. Our results indicate that excess Co ions occupy tetrahedral interstitial sites at lower excess Co stoichiometries, and become substitutional for octahedrally-coordinated Mn at higher Co levels. We compare these results with density functional theory models of stoichiometric CoMn2O4 to understand how the Jahn-Teller distortion and hybridization in Mn-O bonds impact the ability to hole dope the material with excess Co. The findings provide important insights into CoMn2O4 and related spinel oxides that are promising candidates for inexpensive oxygen reduction reaction catalysts.

Electronic and Structural Properties of Single-crystal Jahn-Teller Active Co1+xMn2-xO4Thin Films.

Recent investigations on spinel CoMn2O4have shown its potential for applications in water splitting and fuel cell technologies as it exhibits strong catalytic behavior through oxygen reduction reactivity. To further understand this material, we report for the first time the synthesis of single-crystalline Co1+xMn2-xO4thin films using molecular beam epitaxy. By varying sample composition, we establish links between cation stoichiometry and material properties using in-situ x-ray photoelectron spectroscopy, x-ray diffraction, scanning transmission electron microscopy, x-ray absorption spectroscopy, and spectroscopic ellipsometry. Our results indicate that excess Co ions occupy tetrahedral interstitial sites at lower excess Co stoichiometries, and become substitutional for octahedrally-coordinated Mn at higher Co levels. We compare these results with density functional theory models of stoichiometric CoMn2O4to understand how the Jahn-Teller distortion and hybridization in Mn-O bonds impact the ability to hole dope the material with excess Co. The findings provide important insights into CoMn2O4and related spinel oxides that are promising candidates for inexpensive oxygen reduction reaction catalysts.

Clinical Effectiveness of an Adaptive Treatment Planning Algorithm for Intensity Modulated Radiation Therapy Versus 3D Conformal Radiation Therapy for Node-Positive Breast Cancer Patients Undergoing Regional Nodal Irradiation/Postmastectomy Radiation Therapy.

PURPOSE: Clinical trials support adjuvant regional nodal irradiation (RNI) after breast-conserving surgery or mastectomy for patients with lymph node-positive breast cancer. Advanced treatment planning techniques (eg, intensity modulated radiation therapy [IMRT]) can reduce dose to organs at risk (OARs) in this situation. However, uncertainty persists about when IMRT is clinically indicated (vs 3-dimensional conformal radiation therapy [3DCRT]) for RNI. We hypothesized that an adaptive treatment planning algorithm (TPA) for IMRT adoption would allow OAR constraints for RNI to be met when 3DCRT could not without significantly changing toxicity and locoregional recurrence (LRR) patterns. METHODS AND MATERIALS: Since 2013, all RNI patients also underwent an adaptive TPA that began with 3DCRT and then changed to IMRT when OAR constraints (mean heart dose ≤500 cGy; ipsilateral lung V20 ≤35%) could not be met. Patients received 2 Gy/d to the prospectively contoured target volumes (including internal mammary nodes). We retrospectively evaluated the dosimetry and clinical outcomes of the treatment groups (IMRT vs 3DCRT). The primary endpoint was the cumulative incidence of LRR as the site of first recurrence, and we specifically address patterns of failure based on dose to the posterior supraclavicular nodal region (SCL-post). RESULTS: Two hundred forty patients (60% stage III; mean 4.0 + nodes) underwent an adaptive-TPA for RNI after mastectomy (74%) or breast-conserving surgery (26%), resulting in 168 patients treated with 3DCRT and 72 patients treated with IMRT. There were 7 LRRs (2 IMRT, 5 3DCRT) resulting in 4-year LRR of 2.8% for IMRT versus 1.8% for 3DCRT (P = .99). Three patients (2 IMRT, 1 3DCRT) had SCL nodal failures (1 in the SCL-post). CONCLUSIONS: An adaptive TPA for use of IMRT when 3DCRT does not meet critical OAR constraints resulted in rare high-grade toxicity and no difference in failure patterns between patients treated with IMRT and 3DCRT. These data should provide reassurance that IMRT maintains the therapeutic ratio by preserving cancer control outcomes without excess toxicity when 3DCRT fails to meet OAR constraints.

Implementation of cancer next-generation sequencing testing in a community hospital.

In the current era of personalized medicine, the field of oncology is witnessing a paradigm shift in patient care that is driving a tighter integration of genomic analysis modalities in patient care decision. This is driven by the expanding category of targeted therapies that require a broader understanding of the mutational profile of patient samples to more precisely guided personalized treatment decisions. Next-generation sequencing (NGS) has proved to be of tremendous power in detecting and characterizing a broad spectrum of activating or loss-of-function mutations across many gene targets. This power of NGS also results in significant challenges related to technical expertise, bioinformatics, computing infrastructure, laboratory practices, and integration into clinical decision-making. These challenges are particularly relevant to smaller and mid-tier hospital networks that are faced with the need to modernize their clinical practices and offer their patients access to advanced genomic technologies to improve outcomes. Adoption of such personalized medicine relies on information about a patient's cancer genome and the identification of its variants. This is best achieved using NGS. However, there are challenges to the adoption of such a complex technology and workflow, especially in smaller hospital systems. This commentary summarizes key considerations and challenges related to implementation of NGS in a community hospital setting.

On the Cutting Edge of Research to Conserve At-Risk Species: Maximizing Impact through Partnerships.

Today's conservation challenges are complex. Solving these challenges often requires scientific collaborations that extend beyond the scope, expertise, and capacity of any single agency, organization, or institution. Conservation efforts can benefit from interdisciplinary collaboration, scientific and technological innovations, and the leveraging of capacity and resources among partners. Here we explore a series of case studies demonstrating how collaborative scientific partnerships are furthering the mission of the US Fish and Wildlife Service (USFWS), including: (1) contaminants of emerging concern in the Great Lakes Basin, (2) Poweshiek skipperling conservation, (3) using technology to improve population survey methods for bats and monarch butterfly, and (4) Big River restoration in the Southeast Missouri lead mining district. These case studies illustrate how strategic and effective scientific collaboration is a multi-stage process that requires investment of time and resources by all participants. Early coordination and communication is crucial to aligning planned work with scientific and decision-making needs. Collaborations between USFWS and external scientists can be mutually beneficial by supporting the agency mission while also providing an avenue for innovative research to be directly applied in conservation decisions and management actions.

Race Does Not Affect Tumor Control, Adverse Effects, or Quality of Life after Proton Therapy.

PURPOSE: To compare 5-year biochemical control, toxicity, and patient-reported quality of life (QOL) outcomes for African American and White patients treated with proton therapy (PT) for prostate cancer. MATERIALS AND METHODS: We reviewed the medical records of 1,066 men with clinically localized prostate cancer. Patients were treated with definitive PT between 2006 and 2010. Patients received a median radiation dose of 78 Gy (RBE) with conventional fractionation (1.8- 2 Gy [RBE] per fraction). Sixty-eight (6.4%) men self-identified as African American and 998 (93.6%) self-identified as White. Five-year rates of biochemical control, grade 3 genitourinary and gastrointestinal toxicity, and patient-reported QOL are reported and compared between African American and White patients. RESULTS: Median biochemical follow-up was 5.0 years for both African American and White patients. Median follow-up for toxicity was 5.0 and 5.2 years, respectively. On multivariate analysis, race was not a significant predictor for 5-year freedom from biochemical failure (HR 0.8, p=0.55). No significant association was found between race and grade 3 genitourinary toxicity on multivariate analysis at 5 years (HR 2.5, p=0.10). Patient-reported QOL using median EPIC bowel, urinary incontinence, and irritative summaries scores were not significantly different between the groups. African Americans had higher median sexual summary scores at 2 years than White patients (75 vs. 54, p=0.01) but by 5+ years, the sexual summary scores were no longer significantly different (63 vs. 53, p=0.35). CONCLUSION: With a median follow-up of 5 years, there were no racial disparities in biochemical control, grade 3 toxicity, or patient-reported QOL after PT for prostate cancer.

Five-Year Biochemical Results, Toxicity, and Patient-Reported Quality of Life After Delivery of Dose-Escalated Image Guided Proton Therapy for Prostate Cancer.

PURPOSE: To report clinical outcomes in patients treated with image guided proton therapy (PT) for localized prostate cancer. METHODS AND MATERIALS: The medical records of 1327 men were reviewed. Each man was enrolled on an outcomes tracking study. Dual enrollment on a prospective clinical trial was allowed. Each patient was treated for localized prostate cancer with PT at our institution between 2006 and 2010. Ninety-eight percent of patients received 78 Gy (radiobiological equivalent [RBE]) or higher; 18% received androgen deprivation therapy (ADT). The 5-year freedom from biochemical progression (FFBP), distant metastasis-free survival, and cause-specific survival rates are reported for each risk group. Data on patient-reported quality of life and high-grade toxicities were prospectively collected and reported. A multivariate analysis was performed to identify clinical predictors of biochemical failure and urologic toxicity. RESULTS: The median follow-up time was 5.5 years. The 5-year FFBP rates were 99%, 94%, and 74% in low-risk, intermediate-risk, and high-risk patients, respectively. The actuarial 5-year rates of late grade 3+ Common Terminology Criteria for Adverse Events, version 4.0, gastrointestinal (GI) and genitourinary (GU) toxicity were 0.6% and 2.9%, respectively. Multivariate analysis showed a significant correlation between grade 3+ GU toxicity and pretreatment prostate reductive procedures (P<.0001), prostate volume (P=.0085), pretreatment α-blockers (P=.0067), diabetes (P=.0195), and dose-volume histogram parameters (P=.0208). The median International Prostate Symptom Scores pretreatment scores and scores at 5 years after treatment were 7 and 7, respectively. The mean Expanded Prostate Cancer Index Composite (EPIC) scores significantly declined for sexual summary for patients not receiving ADT (from 67 to 53) between baseline and 5 years. CONCLUSIONS: Image guided PT provided excellent biochemical control rates for patients with localized prostate cancer. The actuarial rates of high-grade toxicity were low after PT. From pretreatment to 5 years of follow-up, a significant decline was found only in mean EPIC sexual summary scores. Prospective clinical studies are needed to determine the comparative effectiveness of PT and other radiation treatment strategies.

Patient-Reported Quality of Life in Men with Transurethral Resection of the Prostate Undergoing Proton Therapy for Management of Prostate Cancer.

PURPOSE: We report on quality of life (QOL) and early toxicity among men with prostate cancer who underwent transurethral resection of the prostate (TURP) before proton therapy. MATERIALS AND METHODS: Between 2006 and 2010, 1,289 patients were treated definitively with proton therapy for prostate cancer at our institution and enrolled on a prospective outcomes-tracking protocol. Ninety-six of the men had received a TURP before proton therapy, while 1,193 men had not. Baseline comorbidities, medications, expanded prostate index composite (EPIC) score, international prostate symptom score (IPSS), and CTCAE vs.3 toxicity assessment were collected prospectively. The Kaplan-Meier product limit method was used to estimate freedom from toxicity. RESULTS: Men who had TURP before proton therapy had lower baseline EPIC scores for urinary incontinence, bowel summary, and sexual summary compared with the non-TURP group, but no significant difference in urinary obstructive score was observed. After controlling for baseline scores, there was no significant difference in bowel summary or sexual summary scores between the two groups over time. There were, however, differences for urinary irritation/obstruction scores and urinary incontinence scores favoring those patients who did not have a TURP-like procedure. Toxicity assessment showed that the 2-year and 5-year rates of grade 3 genitourinary toxicity in the pretreatment TURP group were 12.3% and 17.2%, respectively. CONCLUSIONS: Pretreatment TURP was associated with both a high incidence of physician-assessed toxicity and inferior patient-reported QOL scores both before and after proton therapy treatment. Studies investigating QOL and toxicity after specific prostate cancer therapies should stratify patients by pretreatment TURP. Longer follow-up is needed to confirm if these differences ever resolve.

PAPC mediates self/non-self-distinction during Snail1-dependent tissue separation.

Cleft-like boundaries represent a type of cell sorting boundary characterized by the presence of a physical gap between tissues. We studied the cleft-like ectoderm-mesoderm boundary in Xenopus laevis and zebrafish gastrulae. We identified the transcription factor Snail1 as being essential for tissue separation, showed that its expression in the mesoderm depends on noncanonical Wnt signaling, and demonstrated that it enables paraxial protocadherin (PAPC) to promote tissue separation through two novel functions. First, PAPC attenuates planar cell polarity signaling at the ectoderm-mesoderm boundary to lower cell adhesion and facilitate cleft formation. Second, PAPC controls formation of a distinct type of adhesive contact between mesoderm and ectoderm cells that shows properties of a cleft-like boundary at the single-cell level. It consists of short stretches of adherens junction-like contacts inserted between intermediate-sized contacts and large intercellular gaps. These roles of PAPC constitute a self/non-self-recognition mechanism that determines the site of boundary formation at the interface between PAPC-expressing and -nonexpressing cells.

Insights in public health: Building support for an evidence-based teen pregnancy and sexually transmitted infection prevention program adapted for foster youth.

Hawai'i Youth Services Network (HYSN) was founded in 1980 and is incorporated as a 501(c) (3) organization. HYSN plays a key role in the planning, creation, and funding of local youth services. One of HYSN's focuses is teen pregnancy and sexually transmitted infections (STI) prevention among foster youth. Foster youth are at a greater risk for teen pregnancy and STI due to a variety of complex factors including instability, trauma, and emancipation from the foster care system. This article highlights how HYSN is leveraging both federal and local funding, as well as other resources, in order to implement an evidence-based teen pregnancy and STI prevention program adapted for foster youth.

Associations between fecal indicator bacteria prevalence and demographic data in private water supplies in Virginia.

Over 1.7 million Virginians rely on private water sources to provide household water. The heaviest reliance on these systems occurs in rural areas, which are often underserved with respect to available financial resources and access to environmental health education. This study aimed to identify potential associations between concentrations of fecal indicator bacteria (FIB) (coliforms, Escherichia coli) in over 800 samples collected at the point-of-use from homes with private water supply systems and homeowner-provided demographic data (household income and education). Of the 828 samples tested, 349 (42%) of samples tested positive for total coliform and 55 (6.6%) tested positive for E. coli. Source tracking efforts targeting optical brightener concentrations via fluorometry and the presence of a human-specific Bacteroides marker via quantitative real-time polymerase chain reaction (qPCR) suggest possible contamination from human septage in over 20 samples. Statistical methods implied that household income has an association with the proportion of samples positive for total coliform, though the relationship between education level and FIB is less clear. Further exploration of links between demographic data and private water quality will be helpful in building effective strategies to improve rural drinking water quality.