The health implications of cumulative exposure to contextual (dis)advantage: Methodological and substantive advances from a unique data linkage.

Deleterious neighborhood conditions are associated with poor health, yet the health impact of cumulative lifetime exposure to neighborhood disadvantage is understudied. Using up to five decades of residential histories for 4,177 adult participants in the Survey of Health of Wisconsin (SHOW) and spatio-temporally linked neighborhood conditions, we develop four operational approaches to characterizing cumulative neighborhood (dis)advantage over the life course. We estimated their associations with self-reported general health and compared to estimates using neighborhood (dis)advantage at time of study enrollment. When cumulative exposures were assessed with the most granular temporal scale (Approach 4), neighborhood transport constraints (OR = 1.21, 95% CI: 1.08, 1.36), residential turnover (OR = 1.20, 95% CI: 1.07, 1.34), education deficit (OR = 1.17, 95% CI: 1.04, 1.32), racial segregation (OR = 1.20, 95% CI: 1.04, 1.38) and median household income (OR = 0.85, 95% CI: 0.75, 0.97) were significantly associated with risk of fair or poor health. For composite neighborhood disadvantage, cumulative exposures had a stronger association (OR = 1.05, 95% CI: 1.02, 1.08) than the cross-sectional exposure (OR = 1.03, 95% CI: 1.01, 1.06). Single point-in-time neighborhood measures underestimate the neighborhood and health relationship, underscoring the importance of a life course approach to cumulative exposure measurement.

Circadian disruption, clock genes, and metabolic health.

A growing body of research has identified circadian-rhythm disruption as a risk factor for metabolic health. However, the underlying biological basis remains complex, and complete molecular mechanisms are unknown. There is emerging evidence from animal and human research to suggest that the expression of core circadian genes, such as circadian locomotor output cycles kaput gene (CLOCK), brain and muscle ARNT-Like 1 gene (BMAL1), period (PER), and cyptochrome (CRY), and the consequent expression of hundreds of circadian output genes are integral to the regulation of cellular metabolism. These circadian mechanisms represent potential pathophysiological pathways linking circadian disruption to adverse metabolic health outcomes, including obesity, metabolic syndrome, and type 2 diabetes. Here, we aim to summarize select evidence from in vivo animal models and compare these results with epidemiologic research findings to advance understanding of existing foundational evidence and potential mechanistic links between circadian disruption and altered clock gene expression contributions to metabolic health-related pathologies. Findings have important implications for the treatment, prevention, and control of metabolic pathologies underlying leading causes of death and disability, including diabetes, cardiovascular disease, and cancer.

Linking sequences of exposure to residential (dis)advantage, individual socioeconomic status, and health.

Life course theories suggest that the relationship between residential (dis)advantage and health is best understood by examining the ordering and duration of cumulative exposures across the life course. This study employs sequence and cluster analysis on two decades of residential histories linked to the Survey of the Health of Wisconsin to define typologies of exposure to residential (dis)advantage and use these typologies to predict self-rated fair/poor health. Exposure to residential (dis)advantage is mostly stable across the adult life course and greater disadvantage predicts fair/poor health. Longitudinal exposures to residential (dis)advantage shape health independently of and in tandem with individual-level resources.

Dietary isoleucine content defines the metabolic and molecular response to a Western diet.

The amino acid composition of the diet has recently emerged as a critical regulator of metabolic health. Consumption of the branched-chain amino acid isoleucine is positively correlated with body mass index in humans, and reducing dietary levels of isoleucine rapidly improves the metabolic health of diet-induced obese male C57BL/6J mice. However, it is unknown how sex, strain, and dietary isoleucine intake may interact to impact the response to a Western Diet (WD). Here, we find that although the magnitude of the effect varies by sex and strain, reducing dietary levels of isoleucine protects C57BL/6J and DBA/2J mice of both sexes from the deleterious metabolic effects of a WD, while increasing dietary levels of isoleucine impairs aspects of metabolic health. Despite broadly positive responses across all sexes and strains to reduced isoleucine, the molecular response of each sex and strain is highly distinctive. Using a multi-omics approach, we identify a core sex- and strain- independent molecular response to dietary isoleucine, and identify mega-clusters of differentially expressed hepatic genes, metabolites, and lipids associated with each phenotype. Intriguingly, the metabolic effects of reduced isoleucine in mice are not associated with FGF21 - and we find that in humans plasma FGF21 levels are likewise not associated with dietary levels of isoleucine. Finally, we find that foods contain a range of isoleucine levels, and that consumption of dietary isoleucine is lower in humans with healthy eating habits. Our results demonstrate that the dietary level of isoleucine is critical in the metabolic and molecular response to a WD, and suggest that lowering dietary levels of isoleucine may be an innovative and translatable strategy to protect from the negative metabolic consequences of a WD.

Corrigendum: The effect of low-abundance OTU filtering methods on the reliability and variability of microbial composition assessed by 16S rRNA amplicon sequencing.

[This corrects the article DOI: 10.3389/fcimb.2023.1165295.].

Diverse mechanisms by which chemical pollutant exposure alters gut microbiota metabolism and inflammation.

The human gut microbiome, the host, and the environment are inextricably linked across the life course with significant health impacts. Consisting of trillions of bacteria, fungi, viruses, and other micro-organisms, microbiota living within our gut are particularly dynamic and responsible for digestion and metabolism of diverse classes of ingested chemical pollutants. Exposure to chemical pollutants not only in early life but throughout growth and into adulthood can alter human hosts' ability to absorb and metabolize xenobiotics, nutrients, and other components critical to health and longevity. Inflammation is a common mechanism underlying multiple environmentally related chronic conditions, including cardiovascular disease, multiple cancer types, and mental health. While growing research supports complex interactions between pollutants and the gut microbiome, significant gaps exist. Few reviews provide descriptions of the complex mechanisms by which chemical pollutants interact with the host microbiome through either direct or indirect pathways to alter disease risk, with a particular focus on inflammatory pathways. This review focuses on examples of several classes of pollutants commonly ingested by humans, including (i) heavy metals, (ii) persistent organic pollutants (POPs), and (iii) nitrates. Digestive enzymes and gut microbes are the first line of absorption and metabolism of these chemicals, and gut microbes have been shown to alter compounds from a less to more toxic state influencing subsequent distribution and excretion. In addition, chemical pollutants may interact with or alter the selection of more harmful and less commensal microbiota, leading to gut dysbiosis, and changes in receptor-mediated signaling pathways that alter the integrity and function of the gut intestinal tract. Arsenic, cadmium, and lead (heavy metals), influence the microbiome directly by altering different classes of bacteria, and subsequently driving inflammation through metabolite production and different signaling pathways (LPS/TLR4 or proteoglycan/TLR2 pathways). POPs can alter gut microbial composition either directly or indirectly depending on their ability to activate key signaling pathways within the intestine (e.g., PCB-126 and AHR). Nitrates and nitrites' effect on the gut and host may depend on their ability to be transformed to secondary and tertiary metabolites by gut bacteria. Future research should continue to support foundational research both in vitro, in vivo, and longitudinal population-based research to better identify opportunities for prevention, gain additional mechanistic insights into the complex interactions between environmental pollutants and the microbiome and support additional translational science.

Determinants of per- and polyfluoroalkyl substances (PFAS) exposure among Wisconsin residents.

BACKGROUND: Per- and polyfluoroalkyl substances (PFAS) include thousands of manufactured compounds with growing public health concerns due to their potential for widespread human exposure and adverse health outcomes. While PFAS contamination remains a significant concern, especially from ingestion of contaminated food and water, determinants of the variability in PFAS exposure among regional and statewide populations in the United States remains unclear. OBJECTIVES: The objective of this study was to leverage The Survey of the Health of Wisconsin (SHOW), the only statewide representative cohort in the US, to assess and characterize the variability of PFAS exposure in a general population. METHODS: This study sample included a sub-sample of 605 adult participants from the 2014-2016 tri-annual statewide representative sample. Geometric means for PFOS, PFOA, PFNA, PFHxS, PFPeS, PFHpA, and a summed measure of 38 analyzed serum PFAS were presented by demographic, diet, behavioral, and residential characteristics. Multivariate linear regression was used to determine significant predictors of serum PFAS after adjustment. RESULTS: Overall, higher serum concentrations of long-chain PFAS were observed compared with short-chain PFAS. Older adults, males, and non-Hispanic White individuals had higher serum PFAS compared to younger adults, females, and non-White individuals. Eating caught fish in the past year was associated with elevated levels of several PFAS. DISCUSSION: This is among the first studies to characterize serum PFAS among a representative statewide sample in Wisconsin. Both short- and long-chain serum PFAS were detectable for six prominent PFAS. Age and consumption of great lakes fish were the most significant predictors of serum PFAS. State-level PFAS biomonitoring is important for identifying high risk populations and informing state public health standards and interventions, especially among those not living near known contamination sites.

Participant attrition from statewide, population-based Survey of the Health of Wisconsin into the longitudinal SHOW COVID-19 cohort.

PURPOSE: Longitudinal studies are essential for examining how social and institutional determinants of health, historical and contemporary, affect disparities in COVID-19 related outcomes. The unequal impacts of COVID-19 likely exacerbated selected attrition in longitudinal research. This study examines attrition and survey mode effects in the SHOW COVID-19 study which recruited from a statewide, representative cohort. MATERIALS & METHODS: Participants were recruited from the Survey of the Health of Wisconsin (SHOW) cohort. Online surveys, or phone interviews, were administered at three timepoints during 2020-2021. The surveys captured social, behavioral, and structural determinants of health and the lived experience. Univariate and multivariate logistic regression was used to examine predictors of participation and survey mode effects. RESULTS: A total of 2304 adults completed at least one COVID-19 online survey. Participants were more educated, older, and more likely to be female, married, non-Hispanic, and White compared to non-participants. Phone participants were older, less educated, and more likely be non-White, food insecure, and have co-morbidities compared to online participants. Mode effects were seen with reporting COVID-19 beliefs, loneliness, and anxiety. CONCLUSION: The SHOW COVID-19 cohort offers unique longitudinal data but suffered from selected attrition. Phone interview is an important mode for retention and representation.

Sense of neighborhood belonging and health: geographic, racial, and socioeconomic variation in Wisconsin.

Background: Individuals' sense of belonging (SoB) to their neighborhood is an understudied psychosocial factor that may influence the association between neighborhood characteristics, health, and disparities across socio-demographic groups. Methods: Using 2014-2016 data from the Survey of the Health of Wisconsin (SHOW, N = 1,706), we conduct a detailed analysis of SoB and health in an American context. We construct OLS and logistic regressions estimating belonging's association with general, physical, and mental health. We explore geographic, racial, and socioeconomic variation to understand both the differential distribution of SoB and its heterogeneous relationship with health. Results: A higher SoB is positively associated with better physical, mental, and general health. White participants report higher SoB than Black participants, yet the association between SoB and mental health is strongest among participants of color and urban residents. Conclusion: Sense of belonging to neighborhood significantly predicts many facets of health, with place and individual characteristics appearing to moderate this relationship. Racial, geographic, and socioeconomic disparities in belonging-health associations raise important questions about who benefits from the social, economic, and physical aspects of local communities.

Socioecologic Factors and Racial Differences in Breast Cancer Multigene Prognostic Scores in US Women.

Importance: Disproportionately aggressive tumor biology among non-Hispanic Black women with early-stage, estrogen receptor (ER)-positive breast cancer contributes to racial disparities in breast cancer mortality. It is unclear whether socioecologic factors underlie racial differences in breast tumor biology. Objective: To examine individual-level (insurance status) and contextual (area-level socioeconomic position and rural or urban residence) factors as possible mediators of racial and ethnic differences in the prevalence of ER-positive breast tumors with aggressive biology, as indicated by a high-risk gene expression profile. Design, Setting, and Participants: This retrospective cohort study included women 18 years or older diagnosed with stage I to II, ER-positive breast cancer between January 1, 2007, and December 31, 2015. All data analyses were conducted between December 2022 and April 2023. Main Outcomes and Measures: The primary outcome was the likelihood of a high-risk recurrence score (RS) (≥26) on the Oncotype DX 21-gene breast tumor prognostic genomic biomarker. Results: Among 69 139 women (mean [SD] age, 57.7 [10.5] years; 6310 Hispanic [9.1%], 274 non-Hispanic American Indian and Alaskan Native [0.4%], 6017 non-Hispanic Asian and Pacific Islander [8.7%], 5380 non-Hispanic Black [7.8%], and 51 158 non-Hispanic White [74.0%]) included in our analysis, non-Hispanic Black (odds ratio [OR], 1.33; 95% CI, 1.23-1.43) and non-Hispanic American Indian and Alaska Native women (OR, 1.38; 95% CI, 1.01-1.86) had greater likelihood of a high-risk RS compared with non-Hispanic White women. There were no significant differences among other racial and ethnic groups. Compared with non-Hispanic White patients, there were greater odds of a high-risk RS for non-Hispanic Black women residing in urban areas (OR, 1.35; 95% CI, 1.24-1.46), but not among rural residents (OR, 1.05; 95% CI, 0.77-1.41). Mediation analysis demonstrated that lack of insurance, county-level disadvantage, and urban vs rural residence partially explained the greater odds of a high-risk RS among non-Hispanic Black women (proportion mediated, 17%; P < .001). Conclusions and Relevance: The findings of this cohort study suggest that the consequences of structural racism extend beyond inequities in health care to drive disparities in breast cancer outcome. Additional research is needed with more comprehensive social and environmental measures to better understand the influence of social determinants on aggressive ER-positive tumor biology among racial and ethnic minoritized women from disadvantaged and historically marginalized communities.

Constructing Residential Histories in a General Population-Based Representative Sample.

Research on neighborhoods and health typically measures neighborhood context at a single point in time. However, neighborhood exposures accumulate over the life course, influenced by both residential mobility and neighborhood change, with potential implications for estimating the impact of neighborhoods on health. Commercial databases offer fine-grained longitudinal residential address data that can enrich life-course spatial epidemiology research, and validated methods for reconstructing residential histories from these databases are needed. Our study draws on unique data from a geographically diverse, population-based representative sample of adult Wisconsin residents and the LexisNexis (New York, New York) Accurint, a commercial personal profile database, to develop a systematic and reliable methodology for constructing individual residential histories. Our analysis demonstrated that creating residential histories across diverse geographical contexts is feasible, and it highlights differences in the information obtained from available residential histories by age, education, race/ethnicity, and rural/urban/suburban residency. Researchers should consider potential address data availability and information biases favoring socioeconomically advantaged individuals and their implications for studying health inequalities. Despite these limitations, LexisNexis data can generate varied residential exposure metrics and be linked to contextual data to enrich research into the contextual determinants of health at varied geographic scales.

Associations of gut microbiome richness and diversity with objective and subjective sleep measures in a population sample.

STUDY OBJECTIVES: Alterations in gut microbiota composition have been associated with several conditions, and there is emerging evidence that sleep quantity and quality are associated with the composition of the gut microbiome. Therefore, this study aimed to assess the associations between several measures of sleep and the gut microbiome in a large, population-based sample. METHODS: Data were collected from participants in the Survey of the Health of Wisconsin from 2016 to 2017 (N = 720). Alpha diversity was estimated using Chao1 richness, Shannon's diversity, and Inverse Simpson's diversity. Beta diversity was estimated using Bray-Curtis dissimilarity. Models for each of the alpha-diversity outcomes were calculated using linear mixed effects models. Permutational multivariate analysis of variance tests were performed to test whether gut microbiome composition differed by sleep measures. Negative binomial models were used to assess whether sleep measures were associated with individual taxa relative abundance. RESULTS: Participants were a mean (SD) age of 55 (16) years and 58% were female. The sample was 83% non-Hispanic white, 10.6% non-Hispanic black, and 3.5% Hispanic. Greater actigraphy-measured night-to-night sleep duration variability, wake-after-sleep onset, lower sleep efficiency, and worse self-reported sleep quality were associated with lower microbiome richness and diversity. Sleep variables were associated with beta-diversity, including actigraphy-measured night-to-night sleep duration variability, sleep latency and efficiency, and self-reported sleep quality, sleep apnea, and napping. Relative abundance of several taxa was associated with night-to-night sleep duration variability, average sleep latency and sleep efficiency, and sleep quality. CONCLUSIONS: This study suggests that sleep may be associated with the composition of the gut microbiome. These results contribute to the body of evidence that modifiable health habits can influence the human gut microbiome.

For Submission to the Journal of Community Health: Carbon Monoxide Awareness and Detector Use in the State of Wisconsin.

Carbon monoxide (CO) is a leading cause of poisoning. CO detectors are a known-effective prevention strategy, however, little is known about use of detectors or knowledge of risk. This study assessed awareness of CO poisoning risk, detector laws, and detector use among a statewide sample. Data collected from the Survey of the Health of Wisconsin (SHOW) included a CO Monitoring module added to the in-home interview for 466 participants representing unique households across Wisconsin in 2018-2019. Univariate and multivariable logistic regression models examined associations between demographic characteristics, awareness of CO laws and detector use. Less than half of households had a verified CO detector. Under 46% were aware of the detector law. Those aware had 2.82 greater odds of having a detector in the home compared to those unaware of the law. Lack of CO law awareness may lead to less frequent detector use and result in higher risk of CO poisoning. This highlights the need for CO risk and detector education to decrease poisonings.

Geographic Isolation and Vulnerability Across Peru's Ecological Regions: The Influence of Regional Contexts of Extraction.

Geographically isolated places are often sites of exported environmental risks, intense resource extraction, exploitation and marginalization, and social policy neglect. These conditions create unique challenges related to vulnerability and adaptation that have direct disaster management implications. Our research investigates the relationship between geographic isolation and flood-related social vulnerability across Peru's ecological regions. Ecoregions have different relationships with colonialism and capitalism that shape vulnerability, and we hypothesize that the relationship between vulnerability and geographic isolation varies across ecoregions. Using mapping techniques and spatial regression analysis, we find that relationships between vulnerability and geographic isolation vary regionally, with differences that suggest alignment with regional contexts of extraction. We find notable differences in vulnerability related to public health infrastructure and access to services and between ecoregions with sharply contrasting histories of natural resource extraction and investment and disinvestment.

Change in alcohol consumption during the Covid-19 pandemic and associations with mental health and financial hardship: results from a survey of Wisconsin patients with cancer.

PURPOSE: Alcohol consumption increases health risks for patients with cancer. The Covid-19 pandemic may have affected drinking habits for these individuals. We surveyed patients with cancer to examine whether changes in drinking habits were related to mental health or financial effects of the pandemic. METHODS: From October 2020 to April 2021, adult patients (age 18-80 years at diagnosis) treated for cancer in southcentral Wisconsin were invited to complete a survey. Age-adjusted percentages for history of anxiety or depression, emotional distress, and financial impacts of Covid-19 overall and by change in alcohol consumption (non-drinker, stable, decreased, or increased) were obtained via logistic regression. RESULTS: In total, 1,875 patients were included in the analysis (median age 64, range 19-87 years), including 9% who increased and 23% who decreased drinking. Compared to stable drinkers (32% of sample), a higher proportion of participants who increased drinking alcohol also reported anxiety or depression (45% vs. 26%), moderate to severe emotional distress (61% vs. 37%) and viewing Covid-19 as a threat to their community (67% vs. 55%). Decreased (vs. stable) drinking was associated with higher prevalence of depression or anxiety diagnosis, emotional distress, and negative financial impacts of the pandemic. Compared to non-drinkers (36% of sample), participants who increased drinking were more likely to report emotional distress (61% vs. 48%). CONCLUSIONS: Patients with cancer from Wisconsin who changed their alcohol consumption during the Covid-19 pandemic were more likely to report poor mental health including anxiety, depression, and emotional distress than persons whose alcohol consumption was stable. IMPLICATIONS FOR CANCER SURVIVORS: Clinicians working with cancer survivors should be aware of the link between poor mental health and increased alcohol consumption and be prepared to offer guidance or referrals to counseling, as needed.

Biomonitoring of perfluoroalkyl and polyfluoroalkyl substances (PFAS) from the Survey of the Health of Wisconsin (SHOW) 2014-2016 and comparison with the National Health and Nutrition Examination Survey (NHANES).

BACKGROUND: Per- and polyfluoroalkyl substances (PFAS) are a growing class of manufactured chemical compounds found in a variety of consumer products. PFAS are ubiquitous in the environment and were found in many humans sampled in the United States (U.S.). Yet, significant gaps in understanding statewide levels of exposure to PFAS remain. OBJECTIVE: The goals of this study are to establish a baseline of exposure at the state level by measuring PFAS serum levels among a representative sample of Wisconsin residents and compare to United States National Health and Nutrition Examination Survey (NHANES). METHODS: The study sample included 605 adults (18+ years of age) selected from the 2014-2016 sample of the Survey of the Health of Wisconsin (SHOW). Thirty-eight PFAS serum concentrations were measured using high-pressure liquid chromatography coupled with tandem mass spectrometric detection (HPLC-MS/MS) and geometric means were presented. Weighted geometric mean serum values of eight PFAS analytes from SHOW were compared to U.S. national levels from the NHANES 2015-2016 sample (PFOS, PFOA, PFNA, PFHxS, PFHpS, PFDA, PFUnDA), and the 2017-2018 sample for Me-PFOSA, PFHPS using the Wilcoxon rank-sum test. RESULTS: PFOS, PFHxS, PFHpS, PFDA, PFNA, and PFOA were detected in over 96% of SHOW participants. In general, SHOW participants had lower serum levels across all PFAS when compared to NHANES. Serum levels increased with age and were higher among males and whites. Similar trends were seen in NHANES, except non-whites had higher PFAS levels at higher percentiles in NHANES. IMPACT STATEMENT: The present study conducts biomonitoring of 38 PFAS among representative sample of residents in the state of Wisconsin. Results suggest that while the majority of Wisconsin residents tested have detectable levels of PFAS in their blood serum, they may have a lower body burden of some PFAS compared to a nationally representative sample. Older adults, males, and whites may have a higher body burden of PFAS relative to other groups, both in Wisconsin and the wider United States.

Neighborhood socioeconomic status is associated with low diversity gut microbiomes and multi-drug resistant microorganism colonization.

Social disparities continue to limit universal access to health care, directly impacting both lifespan and quality of life. Concomitantly, the gut microbiome has been associated with downstream health outcomes including the global rise in antibiotic resistance. However, limited evidence exists examining socioeconomic status (SES) associations with gut microbiome composition. To address this, we collected information on the community-level SES, gut microbiota, and other individual cofactors including colonization by multidrug-resistant organisms (MDROs) in an adult cohort from Wisconsin, USA. We found an association between SES and microbial composition that is mediated by food insecurity. Additionally, we observed a higher prevalence of MDROs isolated from individuals with low diversity microbiomes and low neighborhood SES. Our integrated population-based study considers how the interplay of several social and economic factors combine to influence gut microbial composition while providing a framework for developing future interventions to help mitigate the SES health gap.

Sense of community and mental health: a cross-sectional analysis from a household survey in Wisconsin.

BACKGROUND: In the USA, one in five adults live with a mental illness, and researchers have estimated that nearly half of the population will have a mental illness over the course of their lifetime. Research has shown significant associations between social relationships and mental health outcomes at the individual and population levels. This study aims to examine whether sense of community, a type of social capital, is associated with mental health. METHODS: In a cross-sectional analysis, multiple logistic regression models were used to examine whether sense of community was associated with symptoms of depression, anxiety and stress reported over the last week. The analysis used data from the Survey of the Health of Wisconsin collected between 2014 and 2016. A total of 1647 observations are included in the analyses. RESULTS: Compared with those who report a positive sense of community, those with a negative sense of community had a significantly higher odds of reporting depression, anxiety and stress symptoms. Socioeconomic status is negatively associated with depression and anxiety, but not with stress. Women were more likely to experience moderate, severe, or extremely severe anxiety and stress, compared with men. CONCLUSION: This study extends current understanding of health benefits of social capital and found that individuals' sense of community is associated with reduced symptoms of depression, anxiety and stress. Further research examining mechanisms to support improved sense of community and other types of social capital could benefit health equity research.

The effect of low-abundance OTU filtering methods on the reliability and variability of microbial composition assessed by 16S rRNA amplicon sequencing.

PCR amplicon sequencing may lead to detection of spurious operational taxonomic units (OTUs), inflating estimates of gut microbial diversity. There is no consensus in the analytical approach as to what filtering methods should be applied to remove low-abundance OTUs; moreover, few studies have investigated the reliability of OTU detection within replicates. Here, we investigated the reliability of OTU detection (% agreement in detecting OTU in triplicates) and accuracy of their quantification (assessed by coefficient of variation (CV)) in human stool specimens. Stool samples were collected from 12 participants 22-55 years old. We applied several methods for filtering low-abundance OTUs and determined their impact on alpha-diversity and beta-diversity metrics. The reliability of OTU detection without any filtering was only 44.1% (SE=0.9) but increased after filtering low-abundance OTUs. After filtering OTUs with <0.1% abundance in the dataset, the reliability increased to 87.7% (SE=0.6) but at the expense of removing 6.97% reads from the dataset. When filtering was based on individual sample, the reliability increased to 73.1% after filtering OTUs with <10 copies while removing only 1.12% of reads. High abundance OTUs (>10 copies in sample) had lower CV, indicating better accuracy of quantification than low-abundance OTUs. Excluding very low-abundance OTUs had a significant impact on alpha-diversity metrics sensitive to the presence of rare species (observed OTUs, Chao1) but had little impact on relative abundance of major phyla and families and alpha-diversity metrics accounting for both richness and evenness (Shannon, Inverse Simpson). To increase the reliability of microbial composition, we advise removing OTUs with <10 copies in individual samples, particularly in studies where only one subsample per specimen is available for analysis.