Impact of heat-treatment on wastewater analytical parameters.

Raw wastewater analysis is an important step in treatment assessment; however, it is associated with risks of personnel exposure to pathogens. Such risks are enhanced during virus outbreaks, such as the COVID pandemic, and heat-treatment is a commonly used mitigation measure. We examined whether heat-treatment compromises wastewater analytical parameters results. We found that heat-treatment of blackwater at 60 °C for 90 min in capped containers yielded no statistically different values (p > 0.05) for pH, chemical oxygen demand (COD), ammonia (NH3), total nitrogen (TN), total suspended solids (TSS), and turbidity for specimens from three different sources. This heat-treatment inactivated coliform bacteria (>4 log10 reduction) thus compromising the measurement of commonly used fecal contamination indicators. The observation of intact helminth eggs in heat-treated specimens suggests that the helminth egg enumeration assay is not compromised. These findings indicate that heat-treatment for the safe handling of wastewater, as may be needed in future virus outbreaks, does not affect the measurements of many common wastewater physico-chemical properties.

Feasibility of Mild Traumatic Brain Injury Assessment Based on Cardiovascular Response to Postural Change.

OBJECTIVE: To determine the feasibility of short-term cardiovascular responses to postural change as a screening tool for mild traumatic brain injury (mTBI), using heart rate metrics that can be measured with a wearable electrocardiogram sensor. SETTING: Military TBI clinic. DESIGN: Data collected from active-duty service members who had sustained a medically diagnosed mTBI within the prior 72 hours and from age- and sex-matched controls. Cardiac data collected while participants performed a sequence of postural changes. MAIN MEASURES: Model classification compared with clinical mTBI diagnosis. RESULTS: Cardiac biomarkers of mTBI were identified and logistic regression classifiers for mTBI were developed from different subsets of biomarkers. The best model achieved 90% sensitivity and 69% specificity using data from 2 different postural changes. CONCLUSION: Noninvasive measurement of cardiovascular response to postural change is a promising approach for field-deployable post-mTBI screening.

Fecal Malodor Detection Using Low-Cost Electrochemical Sensors.

Technology innovation in sanitation is needed for the 4.2 billion people worldwide, lacking safely managed sanitation services. A major requirement for the adoption of these technologies is the management of malodor around toilet and treatment systems. There is an unmet need for a low-cost instrumented technology for detecting the onset of sanitation malodor and triggering corrective actions. This study combines sensory data with low-cost gas sensor data on malodor emanating from feces. The response of 10 commercial electrochemical gas sensors was collected alongside olfactometric measurements. Odor from fecal specimens at different relevant dilution as well as specimens with pleasant odors as a control were evaluated for a total of 64 responses. Several of the sensors responded positively to the fecal odor, with the formaldehyde, hydrogen sulfide, and ammonia sensors featuring the highest signal to noise ratio. A positive trend was observed between the sensors' responses and the concentration of the odorant and with odor intensity, but no clear correspondence with dilution to threshold (D/T) values was found. Selected sensors were responsive both above and below the intensity values used as the cutoff for offensive odor, suggesting the possibility of using those sensors to differentiate odor offensiveness based just on the magnitude of their response. The specificity of the sensors suggested that discrimination between the selected non-fecal and fecal odors was possible. This study demonstrates that some of the evaluated sensors could be used to assemble a low-cost malodor warning system.

Soil-transmitted helminth eggs assessment in wastewater in an urban area in India.

Water quality and sanitation are inextricably linked to prevalence and control of soil-transmitted helminth infections, a public health concern in resource-limited settings. India bears a large burden of disease associated with poor sanitation. Transformative onsite sanitation technologies are being developed that feature elimination of pathogens including helminth eggs in wastewater treatment. We are conducting third-party testing of multiple sanitation technology systems in Coimbatore (Tamil Nadu) India. To ensure stringent testing of the pathogen removal ability of sanitation technologies, the presence of helminth eggs in wastewater across the town of Coimbatore was assessed. Wastewater samples from existing test sites as well as desludging trucks servicing residential and non-residential septic tanks, were collected. The AmBic methodology (based on washing, sieving, sedimenting and floating) was used for helminth egg isolation. We tested 29 different source samples and found a 52% prevalence of potentially infective helminth eggs. Identification and enumeration of helminth species is reported against the septage source (private residential vs. shared toilet facility) and total solids content. Trichuris egg counts were higher than those of hookworm and Ascaris from desludging trucks, whereas hookworm egg counts were higher in fresh wastewater samples. Surprisingly, no correlation between soil transmitted helminth eggs and total solids was observed.

In vitro exposure systems and dosimetry assessment tools for inhaled tobacco products: Workshop proceedings, conclusions and paths forward for in vitro model use.

In 2009, the passing of the Family Smoking Prevention and Tobacco Control Act facilitated the establishment of the FDA Center for Tobacco Products (CTP), and gave it regulatory authority over the marketing, manufacture and distribution of tobacco products, including those termed 'modified risk'. On 4-6 April 2016, the Institute for In Vitro Sciences, Inc. (IIVS) convened a workshop conference entitled, In Vitro Exposure Systems and Dosimetry Assessment Tools for Inhaled Tobacco Products, to bring together stakeholders representing regulatory agencies, academia and industry to address the research priorities articulated by the FDA CTP. Specific topics were covered to assess the status of current in vitro smoke and aerosol/vapour exposure systems, as well as the various approaches and challenges to quantifying the complex exposures in in vitro pulmonary models developed for evaluating adverse pulmonary events resulting from tobacco product exposures. The four core topics covered were: a) Tobacco Smoke and E-Cigarette Aerosols; b) Air-Liquid Interface-In Vitro Exposure Systems; c) Dosimetry Approaches for Particles and Vapours/In Vitro Dosimetry Determinations; and d) Exposure Microenvironment/Physiology of Cells. The 2.5-day workshop included presentations from 20 expert speakers, poster sessions, networking discussions, and breakout sessions which identified key findings and provided recommendations to advance these technologies. Here, we will report on the proceedings, recommendations, and outcome of the April 2016 technical workshop, including paths forward for developing and validating non-animal test methods for tobacco product smoke and next generation tobacco product aerosol/vapour exposures. With the recent FDA publication of the final deeming rule for the governance of tobacco products, there is an unprecedented necessity to evaluate a very large number of tobacco-based products and ingredients. The questionable relevance, high cost, and ethical considerations for the use of in vivo testing methods highlight the necessity of robust in vitro approaches to elucidate tobacco-based exposures and how they may lead to pulmonary diseases that contribute to lung exposure-induced mortality worldwide.

Reverse genetics with a full-length infectious cDNA of the Middle East respiratory syndrome coronavirus.

Severe acute respiratory syndrome with high mortality rates (~50%) is associated with a novel group 2c betacoronavirus designated Middle East respiratory syndrome coronavirus (MERS-CoV). We synthesized a panel of contiguous cDNAs that spanned the entire genome. Following contig assembly into genome-length cDNA, transfected full-length transcripts recovered several recombinant viruses (rMERS-CoV) that contained the expected marker mutations inserted into the component clones. Because the wild-type MERS-CoV contains a tissue culture-adapted T1015N mutation in the S glycoprotein, rMERS-CoV replicated ~0.5 log less efficiently than wild-type virus. In addition, we ablated expression of the accessory protein ORF5 (rMERS•ORF5) and replaced it with tomato red fluorescent protein (rMERS-RFP) or deleted the entire ORF3, 4, and 5 accessory cluster (rMERS-ΔORF3-5). Recombinant rMERS-CoV, rMERS-CoV•ORF5, and MERS-CoV-RFP replicated to high titers, whereas MERS-ΔORF3-5 showed 1-1.5 logs reduced titer compared with rMERS-CoV. Northern blot analyses confirmed the associated molecular changes in the recombinant viruses, and sequence analysis demonstrated that RFP was expressed from the appropriate consensus sequence AACGAA. We further show dipeptidyl peptidase 4 expression, MERS-CoV replication, and RNA and protein synthesis in human airway epithelial cell cultures, primary lung fibroblasts, primary lung microvascular endothelial cells, and primary alveolar type II pneumocytes, demonstrating a much broader tissue tropism than severe acute respiratory syndrome coronavirus. The availability of a MERS-CoV molecular clone, as well as recombinant viruses expressing indicator proteins, will allow for high-throughput testing of therapeutic compounds and provide a genetic platform for studying gene function and the rational design of live virus vaccines.

High-throughput cardiac safety evaluation and multi-parameter arrhythmia profiling of cardiomyocytes using microelectrode arrays.

Microelectrode arrays (MEAs) recording extracellular field potentials of human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CM) provide a rich data set for functional assessment of drug response. The aim of this work is the development of a method for a systematic analysis of arrhythmia using MEAs, with emphasis on the development of six parameters accounting for different types of cardiomyocyte signal irregularities. We describe a software approach to carry out such analysis automatically including generation of a heat map that enables quick visualization of arrhythmic liability of compounds. We also implemented signal processing techniques for reliable extraction of the repolarization peak for field potential duration (FPD) measurement even from recordings with low signal to noise ratios. We measured hiPS-CM's on a 48 well MEA system with 5minute recordings at multiple time points (0.5, 1, 2 and 4h) after drug exposure. We evaluated concentration responses for seven compounds with a combination of hERG, QT and clinical proarrhythmia properties: Verapamil, Ranolazine, Flecainide, Amiodarone, Ouabain, Cisapride, and Terfenadine. The predictive utility of MEA parameters as surrogates of these clinical effects were examined. The beat rate and FPD results exhibited good correlations with previous MEA studies in stem cell derived cardiomyocytes and clinical data. The six-parameter arrhythmia assessment exhibited excellent predictive agreement with the known arrhythmogenic potential of the tested compounds, and holds promise as a new method to predict arrhythmic liability.

Sensor-based evaluation of a Urine Trap toilet in a shared bathroom.

Urine diversion in a No-Mix Toilet is a promising approach for sustainable fertilizers and reduction of the nutrient load for wastewater treatment; however, user adoption remains a challenge. This study evaluates the Urine Trap, a passive No-Mix toilet design based on the teapot effect, wherein the urine stream inlet is invisible to the user and therefore it does not impact the user experience for increased adoption. This study evaluated the nutrient separation performance of a Urine Trap flush toilet in a bathroom shared by women in two sites in India. Over three different testing periods, 841 uses of this squat plate were recorded in 50 days. Analytical measurements found 36 % separation efficiency for total nitrogen (TN). While effective, the Urine Trap under test by users did not yield a 70-80 % TN separation efficiency observed under engineering characterization. High temporal resolution data from sensors on waste collection tanks, the opening of the bathroom door, and cleansing water flow were used to gain insights into hygiene practices. The data showed a frequent habit of wetting the squat plate during physiological excretion, a hygienic practice that eases cleaning but degrades the teapot separation effect of the Urine Trap design. By using sensors, we demonstrate a method to non-invasively gain quantitative insights into hygiene practices to inform sanitation technologies deployment strategies for improved outcomes.

Electromechanical performance of piezoelectric scanning mirrors for medical endoscopy.

The electromechanical performance of piezoelectric scanning mirrors for endoscopy imaging is presented. The devices are supported by a single actuating cantilever to achieve a high fill factor, the ratio of mirror area to the combined mirror and actuator area. The largest fill factor devices (74%) achieved 10° mechanical scan range at +/-10V with a 300 µm long cantilever. The largest angular displacement of 30° mechanical scan range was obtained with a 500 µm long cantilever device with a 63% fill factor driven at 40 Vpp. A systematic investigation of device performance (displacement and speed) as a function of fabrication and operational parameters including the stress balance in the cantilever revealed unexpectedly large displacements with lack of inversion at the coercive field. An interpretation of the results is presented based on piezoelectric film domain orientation and clamping with supporting piezoelectric film characterization measurements.

Predictive values of time-dense SARS-CoV-2 wastewater analysis in university campus buildings.

Wastewater-based SARS-CoV-2 surveillance on college campuses has the ability to detect individual clinical COVID-19 cases at the building-level. High concordance of wastewater results and clinical cases has been observed when calculated over a time window of four days or longer and in settings with high incidence of infection. At Duke University, twice a week clinical surveillance of all resident undergraduates was carried out in the spring 2021 semester. We conducted simultaneous wastewater surveillance with daily frequency on selected residence halls to assess wastewater as an early warning tool during times of low transmission with the hope of scaling down clinical test frequency. We evaluated the temporal relationship of the two time-dense data sets, wastewater and clinical, and sought a strategy to achieve the highest wastewater predictive values using the shortest time window to enable timely intervention. There were 11 days with clinical cases in the residence halls (80-120 occupants) under wastewater surveillance with 5 instances of a single clinical case and 3 instances of two clinical cases which also corresponded to a positive wastewater SARS-CoV-2 signal. While the majority (71%) of our wastewater samples were negative for SARS-CoV-2, 29% resulted in at least one positive PCR signal, some of which did not correlate with an identified clinical case. Using a criteria of two consecutive days of positive wastewater signals, we obtained a positive predictive value (PPV) of 75% and a negative predictive value of 87% using a short 2 day time window for agreement. A conventional concordance over a much longer 4 day time window resulted in PPV of only 60%. Our data indicated that daily wastewater collection and using a criteria of two consecutive days of positive wastewater signals was the most predictive approach to timely early warning of COVID-19 cases at the building level.

A hands-free stool sampling system for monitoring intestinal health and disease.

Analysis of stool offers simple, non-invasive monitoring for many gastrointestinal (GI) diseases and access to the gut microbiome, however adherence to stool sampling protocols remains a major challenge because of the prevalent dislike of handling one's feces. We present a technology that enables individual stool specimen collection from toilet wastewater for fecal protein and molecular assay. Human stool specimens and a benchtop test platform integrated with a commercial toilet were used to demonstrate reliable specimen collection over a wide range of stool consistencies by solid/liquid separation followed by spray-erosion. The obtained fecal suspensions were used to perform occult blood tests for GI cancer screening and for microbiome 16S rRNA analysis. Using occult blood home test kits, we found overall 90% agreement with standard sampling, 96% sensitivity and 86% specificity. Microbiome analysis revealed no significant difference in within-sample species diversity compared to standard sampling and specimen cross-contamination was below the detection limit of the assay. Furthermore, we report on the use of an analogue turbidity sensor to assess in real time loose stools for tracking of diarrhea. Implementation of this technology in residential settings will improve the quality of GI healthcare by facilitating increased adherence to routine stool monitoring.

Potential Pitfalls in Wastewater Phosphorus Analysis and How to Avoid Them.

Due to the increasing adoption of nutrient discharge regulations, many research groups are stepping into new territory with phosphorus (P) measurements. Accurate reporting of P concentrations in effluent from novel wastewater treatment technologies is critical for protecting both environmental and human health. Analysis of P in wastewater is prone to pitfalls because of the (1) variety of chemical forms of P in wastewater (orthophosphate, condensed P, and organic P), (2) availability of different chemical assays for measuring different P forms, and (3) different conventions in the units for reporting P. Here, we present a case study highlighting how these pitfalls affect analysis and interpretation of P measurements. We show that, when used appropriately, commercially-available kits are indeed accurate tools for evaluating reactive P and total P concentrations. For both standard solutions and real wastewater, we systematically remove steps from the total P protocol to show how protocol deviations affect the results. While standard solutions are important for validating analytical methods, commercially-available wastewater standard solutions only contain P as orthophosphate (reactive P). We therefore demonstrate options for making a mixed-P standard solution containing acid-hydrolyzable and/or organic P compounds that can be used to validate both reactive P and total P assays.

Optimal design of an electrochemical reactor for blackwater treatment.

Electrolysis of blackwater for disinfection and nutrient removal is a portable and scalable technology that can lessen the need for cities to construct large-scale wastewater treatment infrastructure and enable the safe onsite reuse of blackwater. Several systems for treating wastewater from single toilets are described in the literature, but there are few examples of systems designed to use electrolysis to treat blackwater from nearby toilets, which is a situation more common in densely packed urban living environments. In order to scale a single toilet electrolysis system to one that could service multiple toilets, computational fluid dynamic analysis was used to optimize the electrochemical reactor design, and laboratory and field-testing were used to confirm results. Design efforts included optimization of the reactor shape and mixing to improve treatment efficiency, as well as automated cleaning and salt injection to reduce maintenance and service requirements. PRACTITIONER POINTS: Design of a reverse polarity mechanism to enable in situ electrode cleaning and improve long-term electrode performance. Optimization of a hopper design and drainpipe location to collect and remove flaking precipitates and mitigate maintenance issues. Design of an automated salt injection system to guarantee sufficient chloride levels for producing adequate chlorine residuals for consistent disinfection.

Sensor-Array Optimization Based on Time-Series Data Analytics for Sanitation-Related Malodor Detection.

There is an unmet need for a low-cost instrumented technology for detecting sanitation-related malodor as an alert for maintenance around shared toilets and emerging technologies for onsite waste treatment. In this article, our approach to an electronic nose for sanitation-related malodor is based on the use of electrochemical gas sensors, and machine-learning techniques for sensor selection and odor classification. We screened 10 sensors from different vendors with specific target gases and recorded their response to malodor from fecal specimens and urine specimens, and confounding good odors such as popcorn. The analysis of 180 odor exposures data by two feature-selection methods based on mutual information indicates that, for malodor detection, the decision tree (DT) classifier with seven features from four sensors provides 88.0% balanced accuracy and 85.1% macro F1 score. However, the k-nearest-neighbors (KNN) classifier with only three features (from two sensors) obtains 83.3% balanced accuracy and 81.3% macro F1 score. For classification of urine against feces malodor, a balanced accuracy of 94.0% and a macro F1 score of 92.9% are achieved using only four features from three sensors and a logistic regression (LR) classifier.

Field testing of a household-scale onsite blackwater treatment system in Coimbatore, India.

4.2 billion people live without access to safely managed sanitation services. This report describes the field testing of an onsite prototype system designed to treat blackwater from a single flush toilet and reuse of the treated effluent for flushing. The system passes wastewater through a solid-liquid separator followed by settling tanks and granular activated carbon columns into an electrochemical reactor that oxidizes chloride salts from urine to generate chlorine to remove pathogens. The objectives of the study were to verify the functionality of the system (previously demonstrated in the laboratory) under realistic use conditions, to identify maintenance requirements, and to make a preliminary assessment of the system's user acceptability. The prototype was installed in a women's workplace and residential toilet block in Coimbatore, India, and tested over a period of 10 months. The treated water met stringent disinfection threshold for both E. coli and helminth eggs and produced a clear, colorless effluent that met or nearly met local and international discharge standards for non-sewered sanitation systems. The effluent had an average chemical oxygen demand of 81 mg/L, total suspended solids of 11 mg/L, and reduction of total nitrogen by 65%. These tests determined the recommended service lifetimes and maintenance intervals for key system components including the electrochemical cell, granular activated carbon columns, and solid-liquid separator. User feedback regarding the use of treated blackwater as flush water was positive. These findings will inform the design and implementation of next-generation systems currently under development.

Field testing of a household-scale onsite blackwater treatment system in South Africa.

Innovations that enable cost-effective and resource-conserving treatment of human waste are required for the 4.2 billion people in the world who currently lack safe and reliable sanitation services. Onsite treatment and reuse of blackwater is one strategy towards this end, greatly reducing the need to transport wastewater over long distances either via sewers or trucks. Here, we report on the field testing of a prototype onsite blackwater treatment system conducted over a period of 8 months. The system was connected to a women's toilet in a public communal ablution block located in an informal settlement near Durban, South Africa. Liquid waste was treated by separation and diversion of large solids, settling of suspended solids, and filtration through activated carbon prior to disinfection by electrochemical oxidation. System performance was monitored daily by measurement of chemical and physical water quality parameters onsite and confirmed by periodic detailed analysis of chemical and biological parameters at an offsite lab. Daily monitoring of system performance indicated that the effluent had minimal color and turbidity (maximum 90 Pt/Co units and 6.48 NTU, respectively), and consistent evolution of chlorine as blackwater passed through the system. Weekly offsite analysis confirmed that the system consistently inactivated pathogens (E. coli and coliforms) and reduced chemical oxygen demand and total suspended solids to meet ISO 30500 category B standards. Significant reductions in total nitrogen load were also observed, though these reductions often fell short of the 70% reduction required by ISO 30500. No significant reduction in total phosphorus was observed. Maintenance requirements were identified, and the resilience of the system to restart following a prolonged shutdown was demonstrated, but significant improvements are required in the design of the solid/liquid separation mechanism for application of this system in a wiping culture.

Resolving the relative contributions of cistern and pour flushing to toilet water usage: Measurements from urban test sites in India.

A challenge in water reuse for toilet flushing in India and other Asian countries derives from pour flushing practices. It is a common assumption that the amount of pour flushed water used for personal cleansing is small in comparison to the cistern flush volume, however there is a knowledge gap regarding the actual contribution of each water source to the blackwater amount. In this study, digital water meters were used to measure the fraction of water from personal wash tap relative to cistern water that is used for toilet flushing. High temporal resolution measurements were carried in three different urban sites in the city of Coimbatore in the southern Indian state of Tamil Nadu where onsite sanitation treatment prototypes that may provide reclaimed water for cistern flushing are being tested. Data collected over periods of up to 2 months show that the contribution of the cistern flush to the total blackwater volume is low (14-40%). These data highlight an important factor to inform interventions designed around water reuse for flushing in world geographies where personal toilet cleansing by water is the common practice.

An analysis of stereotypical motor movements and cardiovascular coupling in individuals on the autism spectrum.

One of the core diagnostic features of autism spectrum disorder (ASD) is engagement in stereotypical motor movements, although the etiology of this repetitive behavior is unknown. Since the 1960s, it has been hypothesized that stereotypical motor movements serve a homeostatic regulation function, and thereby a putative coupling mechanism to cardiovascular arousal. However, to date, surprisingly few reports explicitly assess cardio-somatic coupling and stereotypical motor movements. The present exploratory study investigates coupling of stereotypical body rocking and hand flapping to heart rate and heart rate variability (HRV) in a convenience sample (n = 10) of children and young adults with moderate to profound ASD. Motor movements were recorded via video and three-axis accelerometry, and simultaneous electrocardiographic signals were obtained to determine cardiovascular indices at or around the onset of naturalistically occurring stereotypy. Analysis of the heart rate revealed both repetitive body rocking and hand flapping in particular were found to associate with a strikingly similar cardiovascular pattern of acceleration and deceleration unrelated to physical demands associated with the movements themselves. Furthermore, neither type of stereotypical movement provoked directional change in heart rate variability. The implications of these results and opportunities for future research are discussed.

Automated Detection of Repetitive Motor Behaviors as an Outcome Measurement in Intellectual and Developmental Disabilities.

Repetitive sensory motor behaviors are a direct target for clinical treatment and a potential treatment endpoint for individuals with intellectual or developmental disabilities. By removing the burden associated with video annotation or direct observation, automated detection of stereotypy would allow for longer term monitoring in ecologic settings. We report automated detection of common stereotypical motor movements using commercially available accelerometers affixed to the body and a generalizable detection algorithm. The method achieved a sensitivity of 80% for body rocking and 93% for hand flapping without individualized algorithm training or foreknowledge of subject's specific movements. This approach is well-suited for implementation in a continuous monitoring system outside of a clinical setting.

Characterization of fecal sludge as biomass feedstock for thermal treatment in the southern Indian state of Tamil Nadu.

Background: Transformative sanitation technologies aim to treat fecal sludge (FS) by thermal processes and recover resources from it. There is a paucity of data describing the relevant properties of FS as viable feedstock for thermal treatment in major geographical target areas, such as India. Methods: This study characterized FS collected from septic tanks in two cities located in the Indian southern state of Tamil Nadu. FS samples were obtained at the point of discharge from trucks in Tiruppur (n=85 samples) and Coimbatore (n=50 samples). Additionally, biosolids obtained from sewage treatment plants (STP) in the cities of Coimbatore and Madurai were characterized. Total solids (TS) were measured, and proximate and ultimate analysis were conducted according to methods used by the fuel industry. Additionally, the ash content was analyzed for heavy metal using standard methods. Results: The average higher heating value (HHV) across all FS samples in Tiruppur (13.4 MJ/kg) was significantly higher than in Coimbatore (5.4 MJ/kg), which was partially attributed to the high ash content of 69% in the latter samples.  The HHV of the biosolids samples ranged from 10 to 12.2 MJ/Kg. The average total solids (TS) content for FS was 3.3% and 2.0% for Tiruppur and Coimbatore respectively, while the median TS content for the two cities was 2.3% and 1.2%. The heavy metal content of the ash was found to be below the thresholds for land disposal. Conclusions: This is one of the first studies that has systematically characterized the calorific and mineral content of septage and biosolids in several cities in India. We expect these data to serve as input data in the design of thermal processes for fecal sludge treatment.