Metagenomic insights into the impact of litter from poultry Concentrated Animal Feeding Operations (CAFOs) to adjacent soil and water microbial communities.

In recent decades, human food consumption has led to an increased demand for animal-based foods, particularly chicken meat production. The state of Georgia, USA is one of the top broiler chicken producers in the United States, where animals are raised in Concentrated Animal Feeding Operations (CAFOs). Without proper management, CAFOs could negatively impact the environment and become a public health risk as a source of water and air pollution and/or by spreading antimicrobial resistance genes. In this study, we used metagenome sequencing to investigate the impact of the application of the CAFO's litter on adjacent soils and downstream creek waters in terms of microbial diversity and antimicrobial resistance profile changes. Our data indicate that while a few microbial groups increased in abundance within a short period of time after litter application, these populations subsequently decreased to levels similar to those found prior to the litter application or to below the detection limit of our metagenome sequencing effort. Microbial taxonomic composition analyses, relative abundance of Metagenome-Assembled Genomes (MAGs) and detection of Antimicrobial Resistance Genes (ARGs) allow us to conclude that this practice of litter application had a negligible effect on the microbiome or resistome profile of these soils and nearby waterways, likely due to its dilution in the field and/or outcompetition by indigenous microbes, revealing a minimal impact of these poultry facilities on the natural microbial communities.

Simultaneous detection and quantification of multiple pathogen targets in wastewater.

Wastewater-based epidemiology has emerged as a critical tool for public health surveillance, building on decades of environmental surveillance work for pathogens such as poliovirus. Work to date has been limited to monitoring a single pathogen or small numbers of pathogens in targeted studies; however, few studies consider simultaneous quantitative analysis of a wide variety of pathogens, which could greatly increase the utility of wastewater surveillance. We developed a novel quantitative multi-pathogen surveillance approach (35 pathogen targets including bacteria, viruses, protozoa, and helminths) using TaqMan Array Cards (TAC) and applied the method on concentrated wastewater samples collected at four wastewater treatment plants in Atlanta, GA from February to October of 2020. From sewersheds serving approximately 2 million people, we detected a wide range of targets including many we expected to find in wastewater (e.g., enterotoxigenic E. coli and Giardia in 97% of 29 samples at stable concentrations) as well as unexpected targets including Strongyloides stercoralis (a human threadworm rarely observed in the USA). Other notable detections included SARS-CoV-2, but also several pathogen targets that are not commonly included in wastewater surveillance like Acanthamoeba spp., Balantidium coli, Entamoeba histolytica, astrovirus, norovirus, and sapovirus. Our data suggest broad utility in expanding the scope of enteric pathogen surveillance in wastewaters, with potential for application in a variety of settings where pathogen quantification in fecal waste streams can inform public health surveillance and selection of control measures to limit infections.

Long-term impacts of an urban sanitation intervention on enteric pathogens in children in Maputo city, Mozambique: study protocol for a cross-sectional follow-up to the Maputo Sanitation (MapSan) trial 5 years postintervention.

INTRODUCTION: We previously assessed the effect of an onsite sanitation intervention in informal neighbourhoods of urban Maputo, Mozambique on enteric pathogen detection in children after 2 years of follow-up (Maputo Sanitation (MapSan) trial, ClinicalTrials.gov: NCT02362932). We found significant reductions in Shigella and Trichuris prevalence but only among children born after the intervention was delivered. In this study, we assess the health impacts of the sanitation intervention after 5 years among children born into study households postintervention. METHODS AND ANALYSIS: We are conducting a cross-sectional household study of enteric pathogen detection in child stool and the environment at compounds (household clusters sharing sanitation and outdoor living space) that received the pour-flush toilet and septic tank intervention at least 5 years prior or meet the original criteria for trial control sites. We are enrolling at least 400 children (ages 29 days to 60 months) in each treatment arm. Our primary outcome is the prevalence of 22 bacterial, protozoan, and soil transmitted helminth enteric pathogens in child stool using the pooled prevalence ratio across the outcome set to assess the overall intervention effect. Secondary outcomes include the individual pathogen detection prevalence and gene copy density of 27 enteric pathogens (including viruses); mean height-for-age, weight-for-age, and weight-for-height z-scores; prevalence of stunting, underweight, and wasting; and the 7-day period prevalence of caregiver-reported diarrhoea. All analyses are adjusted for prespecified covariates and examined for effect measure modification by age. Environmental samples from study households and the public domain are assessed for pathogens and faecal indicators to explore environmental exposures and monitor disease transmission. ETHICS AND DISSEMINATION: Study protocols have been reviewed and approved by human subjects review boards at the Ministry of Health, Republic of Mozambique and the University of North Carolina at Chapel Hill. Deidentified study data will be deposited at https://osf.io/e7pvk/. TRIAL REGISTRATION NUMBER: ISRCTN86084138.

Risk factors for early childhood growth faltering in rural Cambodia: a cross-sectional study.

OBJECTIVE: This study aimed to determine risk factors of growth faltering by assessing childhood nutrition and household water, sanitation, and hygiene (WASH) variables and their association with nutritional status of children under 24 months in rural Cambodia. DESIGN: We conducted surveys in 491 villages (clusters) randomised across 55 rural communes in Cambodia in September 2016 to measure associations between child, household and community-level risk factors for stunting and length-for-age z-score (LAZ). We measured 4036 children under 24 months of age from 3877 households (491 clusters). We analysed associations between nutrition/WASH practices and child growth (LAZ, stunting) using generalised estimating equations (GEEs) to fit linear regression models with robust SEs in a pooled analysis and in age-stratified analyses; child-level and household-level variables were modelled separately from community-level variables. RESULTS: After adjustment for potential confounding, we found household-level and community-level water, sanitation and hygiene factors to be associated with child growth among children under 24 months: presence of water and soap at a household's handwashing station was positively associated with child growth (adjusted mean difference in LAZ +0.10, 95% CI 0.03 to 0.16); household-level use of an improved drinking water source and adequate child stool disposal practices were protective against stunting (adjusted prevalence ratio (aPR) 0.80, 95% CI 0.67 to 0.97; aPR 0.82, 95% CI 0.64 to 1.03). In our age-stratified analysis, we found associations between child growth and community-level factors among children 1-6 months of age: shared sanitation was negatively associated with growth (-0.47 LAZ, 95% CI -0.90 to -0.05 compared with children in communities with no shared facilities); improved sanitation facilities were protective against stunting (aPR 0.43, 95% CI 0.21 to 0.88 compared with children in communities with no improved sanitation facilities); and open defecation was associated with more stunting (aPR 2.13, 95% CI 1.10 to 4.11 compared with children in communities with no open defecation). These sanitation risk factors were only measured in the youngest age strata (1-6 months). Presence of water and soap at the household level were associated with taller children in the 1-6 month and 6-12 month age strata (+0.10 LAZ, 95% CI -0.02 to 0.22 among children 1-6 months of age; +0.11 LAZ, 95% CI -0.02 to 0.25 among children 6-12 months of age compared with children in households with no water and soap). Household use of improved drinking water source was positively associated with growth among older children (+0.13 LAZ, 95% CI -0.01 to 0.28 among children 12-24 months of age). CONCLUSION: In rural Cambodia, water, sanitation and hygiene behaviours were associated with growth faltering among children under 24 months of age. Community-level sanitation factors were positively associated with growth, particularly for infants under 6 months of age. We should continue to make effort to: investigate the relationships between water, sanitation, hygiene and human health and expand WASH access for young children.

A greater awareness of children with glucose-6-phosphate dehydrogenase deficiency is imperative in western countries.

AIM: This national retrospective Danish study described the characteristics of children diagnosed with glucose-6-phosphate dehydrogenase (G6PD) deficiency, an inherited X-linked recessive disorder that often affects children of Middle Eastern descent. METHODS: We studied children born between 1 January 2000 and 31 December 2017 and diagnosed with G6PD deficiency. They were identified from the Danish National Hospital Discharge Register and the Danish Database of Extreme Neonatal Hyperbilirubinaemia. RESULTS: There were 113 children diagnosed with G6PD deficiency, 67% were of Middle Eastern descent and they were frequently diagnosed before the onset of symptoms, based on known heredity. Of the 67 infants born in Denmark, 10% had extreme neonatal hyperbilirubinaemia and one developed kernicterus spectrum disorder, as did one child born in the Middle East. Most (61%) of the 33 children with jaundice received phototherapy, 12% had exchange transfusions and 18% received whole blood transfusions. After the neonatal period, 23% of the cohort had blood transfusions and 4% needed intensive care for acute haemolytic anaemia. The incidence of G6PD deficiency appeared to be severely underestimated. CONCLUSION: Many families from countries where G6PD deficiency is endemic move to Denmark and other Western countries. Greater awareness is essential to avoid chronic and potentially lethal, consequences.

Role of ferroelectric polarization during growth of highly strained ferroelectric materials.

In ferroelectric thin films and superlattices, the polarization is intricately linked to crystal structure. Here we show that it can also play an important role in the growth process, influencing growth rates, relaxation mechanisms, electrical properties and domain structures. This is studied by focusing on the properties of BaTiO3 thin films grown on very thin layers of PbTiO3 using x-ray diffraction, piezoforce microscopy, electrical characterization and rapid in-situ x-ray diffraction reciprocal space maps during the growth using synchrotron radiation. Using a simple model we show that the changes in growth are driven by the energy cost for the top material to sustain the polarization imposed upon it by the underlying layer, and these effects may be expected to occur in other multilayer systems where polarization is present during growth. This motivates the concept of polarization engineering as a complementary approach to strain engineering.

Two Sides of the Same Coin: Environmental and Health Concern Pathways Toward Meat Consumption.

The dramatic increase of meat production in the last decades has proven to be one of the most impacting causes of negative environmental outcomes (e.g., increase of greenhouse emissions, pollution of land and water, and biodiversity loss). In two studies, we aimed to verify the role of key socio-psychological dimensions on meat intake. Study 1 (N = 198) tested the predictive power of an extended version of the Value-Belief-Norm (VBN) model on individual food choices in an online supermarket simulation. In an online survey, participants were directed to a virtual shop and asked to buy food within a set amount of money. Subsequently, they completed measures of behavioral intention, the VBN constructs (values, general pro-environmental beliefs, awareness of consequences, ascription of responsibility, and personal norm), and social norms (injunctive and descriptive). The outcome variable was operationalized in terms of percentage of expenses dedicated to meat and processed meat items, which provided a more robust behavioral measure than the common self-reported ones. Results confirmed the VBN sequential path, showing direct effects of biospheric values and descriptive norm on personal norm. Furthermore, a proof of validity for the new behavioral measure was provided (medium-sized correlation with behavioral intention). Study 2 (N = 218) aimed at verifying whether the meat consumption could be also motivated by a health concern, reflecting individual (cost/benefit) considerations, besides pro-environmental drivers. Results showed the direct impact of health concern and confirmed the indirect role of biospheric values and descriptive norm (via personal norm) on meat intake. This evidence would suggest the use of multiple-frame messages, highlighting both pro-environmental and health consequences, for meat consumption reduction. Nevertheless, the different implications of moral (e.g., environmental concern) vs. non-moral motivators (e.g., health concern) for reducing meat intake need to be stressed: indeed, the first drivers are more central for self-identity and for engaging in environmental citizenship behaviors.

Gut carriage of antimicrobial resistance genes among young children in urban Maputo, Mozambique: Associations with enteric pathogen carriage and environmental risk factors.

Because poor sanitation is hypothesized as a major direct and indirect pathway of exposure to antimicrobial resistance genes (ARGs), we sought to determine a) the prevalence of and b) environmental risk factors for gut carriage of key ARGs in a pediatric cohort at high risk of enteric infections due to poor water, sanitation, and hygiene (WASH) conditions. We investigated ARGs in stool from young children in crowded, low-income settlements of Maputo, Mozambique, and explored potential associations with concurrent enteric pathogen carriage, diarrhea, and environmental risk factors, including WASH. We collected stool from 120 children <14 months old and tested specimens via quantal, multiplex molecular assays for common bacterial, viral, and protozoan enteric pathogens and 84 ARGs encoding potential resistance to 7 antibiotic classes. We estimated associations between ARG detection (number and diversity detected) and concurrently-measured enteric pathogen carriage, recently-reported diarrhea, and risk factors in the child's living environment. The most commonly-detected ARGs encoded resistance to macrolides, lincosamides, and streptogramins (100% of children); tetracyclines (98%); ß-lactams (94%), aminoglycosides (84%); fluoroquinolones (48%); and vancomycin (38%). Neither concurrent diarrhea nor measured environmental (including WASH) conditions were associated with ARG detection in adjusted models. Enteric pathogen carriage and ARG detection were associated: on average, 18% more ARGs were detected in stool from children carrying bacterial pathogens than those without (adjusted risk ratio (RR): 1.18, 95% confidence interval (CI): 1.02, 1.37), with 16% fewer ARGs detected in children carrying parasitic pathogens (protozoans, adjusted RR: 0.84, 95% CI: 0.71, 0.99). We observed gut ARGs conferring potential resistance to a range of antibiotics in this at-risk cohort that had high rates of enteric infection, even among children <14 months-old. Gut ARGs did not appear closely correlated with WASH, though environmental conditions were generally poor. ARG carriage may be associated with concurrent carriage of bacterial enteric pathogens, suggesting indirect linkages to WASH that merit further investigation.

Residential Place Attachment as an Adaptive Strategy for Coping With the Reduction of Spatial Abilities in Old Age.

This study intended to test whether attachment to one's own residential place at neighborhood level could represent a coping response for the elderly (consistently with the "docility hypothesis;" Lawton, 1982), when dealing with the demands of unfamiliar environments, in order to balance their reduction of spatial abilities. Specifically, a sequential path was tested, in which neighborhood attachment was expected to play a buffer role between lowered spatial competence and neighborhood satisfaction. The participants (N = 264), senior citizens (over 65-year-old), responded to a questionnaire including the measures of spatial self-efficacy, spatial anxiety, attitude toward wayfinding, residential attachment and residential satisfaction. Results from the mediation analysis showed that a lower perceived spatial self-efficacy is associated to a higher spatial anxiety, and both promote a more negative attitude toward wayfinding tasks in non-familiar places. This leads to a higher attachment to one's own neighborhood, which in turn predicts a higher residential satisfaction. Thus, the "closure" response of becoming more attached to their residential place may be an adaptive strategy of the elderly for compensating the Person-Environment (P-E) mis-fit (Lawton and Nahemow, 1973) when they feel unable (or less able) to cope with the demands of unfamiliar environments.

Estimation of global recoverable human and animal fecal biomass.

Human and animal feces present persistent threats to global public health and also opportunities for recovery of resources. We present the first global-scale accounting of recoverable feces (livestock animal and human) -from 2003-2030-using country-specific human and animal population estimates and estimated species-specific feces production by human or animal body mass. We also examine global coverage of domestic livestock animals and sanitation facilities to describe the distribution of onsite vs. offsite hazards from animal and human feces. In 2014, the total mass of feces was 3.9 trillion kg/year, increasing by >52 billion kg/year since 2003 and anticipated to reach at least 4.6 trillion kg in 2030. Annual global production of feces from animals-primarily cattle, chickens, and sheep-was about four times that from humans, and ratios of animal:human feces continue to increase (geometric mean of 4.2:1 (2003) vs. 5.0:1 (2014) vs. a projected 6.0:1 (2030)). Low-income populations bear the greatest burden of onsite feces, mostly from animals in or near the domestic environment. This analysis highlights the challenges of resource recovery from concentrated and dispersed sources of feces, and the global public health policy need for safe management of animal feces.

Limited sampling strategy for cyclosporine (Neoral) area under the curve monitoring in pediatric kidney transplant recipients.

Cyclosporine (CSA; Neoral) is one of the most common immunosuppressants used in pediatric renal transplantation. Research in adult renal transplant recipients has shown that 2-h post-dose concentration (C2) monitoring and limited sampling strategies (LSSs) are better at predicting drug exposure and outcome than trough concentrations (C0). While C0 monitoring is the usual practice in pediatric renal transplant patients, area under the curve (AUC) monitoring has been shown to be superior in terms of predictive ability and outcomes. However, AUC monitoring is impractical and inconvenient in a clinic setting because it involves many blood samples. An LSS provides a reliable alternative. The purpose of this study was to prospectively define an LSS (AUC(0-12)) for CSA monitoring and to test its predictive performance. As well, an LSS (AUC(0-4)) for CSA was developed and its predictive performance tested. Blood samples for CSA concentrations were collected in 29 stable pediatric renal transplant patients prior to (t = 0) and at 0.5, 1, 2, 4, 6, and 8 h following a steady-state morning CSA dose. AUC was calculated by the trapezoidal method; LSSs for AUC(0-12) and AUC(0-4) were determined using multiple regression analysis in 14 patients; and the LSSs' predictive performance was tested in 15 additional patients. Both LSSs require two blood samples. For the LSS (AUC(0-12)), blood samples are required immediately before the dose and 2 h post-dose: AUC(0-12) = 12.45 C0 + 2.17 C2 + 723.16 (r2 = 0.909). For the LSS (AUC(0-4)), blood samples are required at one and 2 h post-dose, AUC(0-4) = 1.17 C1 + 1.85 C2 - 41.00 (r2 = 0.971). The LSSs demonstrated low bias and high precision for both AUC(0-12) and AUC(0-4). Our two-concentration LSSs are accurate and precise predictors that are more clinically useful for our patient population than other LSSs that have been developed for pediatric renal transplant patients. Our study template provides a guide for other centers to develop accurate and precise LSSs specific to their own patient population.