Elucidating the role of extracellular polymeric substances (EPS) in dewaterability of fecal sludge from onsite sanitation systems, and changes during anaerobic storage.

As the importance of fecal sludge management (FSM) is increasingly being realized, the need for adequately designed and functioning fecal sludge (FS) treatment plants is also increasing. Research to fill this gap is only emerging and dewatering is a key challenge for developing sustainable treatment solutions. This study evaluated the effect of extracellular polymeric substances (EPS) on dewaterability of FS, and how EPS and dewaterability change during anaerobic storage (as a proxy for time in onsite containment). EPS was extracted from FS and activated sludge using Na2CO3 and sonication and added to sludge samples to determine the effect on dewaterability. The results confirmed that an increase in EPS had a direct impact of decreasing FS dewaterability (as capillary suction time). In this context, we evaluated FS degradation during anaerobic storage, the effect of anaerobic storage time on EPS, EPS fractions and particle size distribution, and the effect of variations in these factors on FS dewaterability. Variations in EPS, EPS fraction and particle size distribution during anaerobic storage were less than expected and average VS reduction of 20% was recorded over 7 weeks. Although anaerobic digestion was verified (biogas production), the results indicate that kinetics of degradation of FS is different from wastewater sludges. Comparatively, EPS fractions in FS were 70 - 75% lower and with higher fractions of humic-like substances than wastewater sludges. Although EPS significantly affects FS dewaterability, anaerobic storage time is not a predictor of dewaterability.

Permeable membrane dewatering of faecal sludge from pit latrines at a transfer station in Nairobi, Kenya.

Faecal sludge dewatering is often a necessary step in many treatment and reuse processes, and it can decrease costs of transporting large volumes of high water content sludge. Common dewatering methods such as settling-thickening tanks and drying beds are not feasible in space-constrained areas such as urban informal settlements, therefore innovative dewatering methods are needed. This study evaluated permeable membranes as a dewatering method for a transfer station operated by Sanergy in Nairobi, Kenya. The objective was to determine if membranes could provide a sustainable and economically-feasible dewatering method within the constraints of Nairobi's informal settlements by evaluating multiple variables, such as flocculant dosage and initial total solids content, and their relation to faecal sludge dewatering. Experiments were conducted with several membrane types including commercial geotextiles and locally-produced plastic woven bags (gunny sacks). Increasing the flocculant dosage within an optimal range was seen to increase dewatering rates, but had minimal effect on the final dewatered solids content. Sludge with initial total solids contents below 3% was seen to require longer dewatering times to allow for drainage, while sludge above 3% required a minimal drainage period before evaporation began. A settling step proved paramount for sludge with high water content where the supernatant was then decanted prior to dewatering through permeable membranes; however, this may not be recommended in all cases depending on the treatment objectives. Reuse of geotextile membranes may lead to a reduction in dewatering performance, and single-use, locally-produced membranes are a more cost-effective, though operationally intensive, option.

Predictive models using "cheap and easy" field measurements: Can they fill a gap in planning, monitoring, and implementing fecal sludge management solutions?

The characteristics of fecal sludge delivered to treatment plants are highly variable. Adapting treatment process operations accordingly is challenging due to a lack of analytical capacity for characterization and monitoring at many treatment plants. Cost-efficient and simple field measurements such as photographs and probe readings could be proxies for process control parameters that normally require laboratory analysis. To investigate this, we evaluated questionnaire data, expert assessments, and simple analytical measurements for fecal sludge collected from 421 onsite containments. This data served as inputs to models of varying complexity. Random forest and linear regression models were able to predict physical-chemical characteristics including total solids (TS) and ammonium (NH4+-N) concentrations, and solid-liquid separation performance including settling efficiency and filtration time (R2 from 0.51-0.66) based on image analysis of photographs (sludge color, supernatant color, and texture) and probe readings (conductivity (EC) and pH). Supernatant color was the best predictor of settling efficiency and filtration time, EC was the best predictor of NH4+-N, and texture was the best predictor of TS. Predictive models have the potential to be applied for real-time monitoring and process control if a database of measurements is developed and models are validated in other cities. Simple decision tree models based on the single classifier of containment type can also be used to make predictions about citywide planning, where a lower degree of accuracy is required.

Evaluation of conceptual model and predictors of faecal sludge dewatering performance in Senegal and Tanzania.

Unpredictable dewatering performance is a barrier to the effective management and treatment of faecal sludge. While mechanisms of dewatering in sludges from wastewater treatment are well understood, it is not clear how dewatering of faecal sludge fits into the framework of existing knowledge. We evaluate physical-chemical parameters, including EPS and cations, and demographic (source), environmental (microbial community), and technical factors (residence time) as possible predictors of dewatering performance in faecal sludge, and make comparisons to the existing conceptual model for wastewater sludge. Faecal sludge from public toilets took longer to dewater than sludge from other sources, and had turbid supernatant after settling. Slow dewatering and turbid supernatant corresponded to high EPS and monovalent cation concentrations, conductivity, and pH, but cake solids after dewatering was not correlated with EPS or other factors. Faecal sludges with higher EPS appeared less stabilised than those with lower EPS, potentially a result of inhibition of biological degradation due to high urine concentrations. However, distinct microbial community compositions were also observed in samples with higher and lower EPS concentrations. Higher EPS faecal sludge was comparable in dewatering behaviour and EPS content to anaerobically digested and primary wastewater sludges. However lower EPS faecal sludges had different dewatering behaviour than wastewater sludges and may be governed by different mechanisms.

Methods to reliably estimate faecal sludge quantities and qualities for the design of treatment technologies and management solutions.

Sanitation access in urban areas of low-income countries is provided through unstandardized onsite technologies containing accumulated faecal sludge. The demand for infrastructure to manage faecal sludge is increasing, however, no reliable method exists to estimate total accumulated quantities and qualities (Q&Q) This proposed approach averages out complexities to estimate conditions at a centralized to semi-centralized scale required for management and treatment technology solutions, as opposed to previous approaches evaluating what happens in individual containments. Empirical data, demographic data, and questionnaires were used in Kampala, Uganda to estimate total faecal sludge accumulation in the city, resulting in 270 L/cap∙year for pit latrines and 280 L/cap∙year for septic tanks. Septic tank sludge was more dilute than pit latrine sludge, however, public toilet was not a distinguishing factor. Non-household sources of sludge represent a significant fraction of the total and have different characteristics than household-level sludge. Income level, water connection, black water only, solid waste, number of users, containment volume, emptying frequency, and truck size were predictors of sludge quality. Empirical relationships such as a COD:TS of 1.09 ±â€¯0.56 could be used for more resource efficient sampling campaigns. Based on this approach, spatially available demographic, technical and environmental (SPA-DET) data and statistical relationships between parameters could be used to predict Q&Q of faecal sludge.

Review of synthetic human faeces and faecal sludge for sanitation and wastewater research.

Investigations involving human faeces and faecal sludge are of great importance for urban sanitation, such as operation and maintenance of sewer systems, or implementation of faecal sludge management. However, working with real faecal matter is difficult as it not only involves working with a pathogenic, malodorous material but also individual faeces and faecal sludge samples are highly variable, making it difficult to execute repeatable experiments. Synthetic faeces and faecal sludge can provide consistently reproducible substrate and alleviate these challenges. A critical literature review of simulants developed for various wastewater and faecal sludge related research is provided. Most individual studies sought to develop a simulant representative of specific physical, chemical, or thermal properties depending on their research objectives. Based on the review, a suitable simulant can be chosen and used or further developed according to the research needs. As an example, the authors present such a modification for the development of a simulant that can be used for investigating the motion (movement, settling and sedimentation) of faeces and their physical and biological disintegration in sewers and in on-site sanitation systems.

Fecal sludge as a fuel: characterization, cofire limits, and evaluation of quality improvement measures.

In many low-income cities, a high proportion of fecal sludge, the excreta and blackwater collected from onsite sanitation systems such as pit latrines, is not safely managed. This constitutes a major danger to environmental and human health. The water, sanitation, and hygiene sector has recognized that valorization of treated fecal sludge could offset the upfront cost of treatment by using it as a fuel source. The few quantitative studies on fecal sludge fuel published to date have focused on heating value, moisture, ash fraction, and heavy metals. However, other factors impacting fuel utility, specifically ash speciation, have not been adequately quantified for fecal sludge. This study contributes to closing that gap and shows the value of more detailed quantification. It first characterizes fecal sludge samples from Colorado and Uganda, confirms that the fuel is better if cofired with other biomass, and outlines a framework for determining safe cofire ratios. Second, the study evaluates two methods for improving fecal sludge as a fuel: carbonization and ash leaching. Carbonization of fecal sludge did not improve fuel quality, but leaching showed promise in ash reduction.

Evaluation of solid fuel char briquettes from human waste.

The developing world faces dual crises of escalating energy demand and lack of urban sanitation infrastructure that pose significant burdens on the environment. This article presents results of a study evaluating the feasibility of using human feces-derived char as a solid fuel for heating and cooking and a potential way to address both crises. The study determined the energy content and the elemental composition of chars pyrolyzed at 300, 450, and 750 °C. Fecal chars made at 300 °C were found to be similar in energy content to wood chars and bituminous coal, having a heating value of 25.6 ± 0.08 MJ/kg, while fecal chars made at 750 °C had an energy content of 13.8 ± 0.48 MJ/kg. The higher heating values of the studied chars were evaluated using their elemental composition and a published predictive model; results found good agreement between the measured and predicted values. Fecal chars made at low temperatures were briquetted with molasses/lime and starch binders. Briquettes made with 10% starch had an average impact resistance index of 79 and a higher heating value of 25 MJ/kg. These values are comparable to those of commercial charcoal briquettes, making fecal char briquettes a potential substitute that also contributes to the preservation of the environment.

Rho Kinase-Mediated Reduction in Cardiac Capillary Endothelial Cell Dimensions, In Situ, Against Flow.

OBJECTIVE: We previously showed that ischemia, induced by interrupting vascular flow, reduced cardiac capillary caliber. This was accomplished by a reduction in endothelial cell dimensions which was sensitive to Rho kinase (ROK) inhibition and stabilization of the actin cytoskeleton. Here, we investigated whether similar changes in endothelial cells, in situ, could be elicited in the presence of flow through the capillary bed. METHODS: Langendorffs perfused rat hearts were subjected to vasoactive agents, ischemia, and reperfusion. Luminal and abluminal perimeters of capillary cross-sections and their areas were measured from electron micrographs to monitor changes in endothelial cell dimensions. RESULTS: Histamine (100 microM) reduced capillary endothelial cell dimensions, in situ, without endothelial injury. While cross-sectional areas of endothelial cells were not altered by histamine, all other parameters measured were significantly reduced in comparison to controls. These changes were pre- vented by ROK inhibition. CONCLUSIONS: Cardiac capillary endothelial cells, in situ, are able to change shape against continuous flow. One hundred (100) microM histamine induces morphometric changes in these endothelial cells, in situ, without cell damage. These cell-shape changes require ROK and mimic those observed following myocardial ischemia. Targeting the actomyosin contractile system may be useful in ameliorating effects of ischemia on the myocardium

Attenuation of glucocorticoid functions in an Anx-A1-/- cell line.

The Ca(2+)- and phospholipid-binding protein Anx-A1 (annexin 1; lipocortin 1) has been described both as an inhibitor of phospholipase A(2) (PLA(2)) activity and as a mediator of glucocorticoid-regulated cell growth and eicosanoid generation. Here we show that, when compared with Anx-A1(+/+) cells, lung fibroblast cell lines derived from the Anx-A1(-/-) mouse exhibit an altered morphology characterized by a spindle-shaped appearance and an accumulation of intracellular organelles. Unlike their wild-type counterparts, Anx-A1(-/-) cells also overexpress cyclo-oxygenase 2 (COX 2), cytosolic PLA(2) and secretory PLA(2) and in response to fetal calf serum, exhibit an exaggerated release of eicosanoids, which is insensitive to dexamethasone (10(-8)- 10(-6) M) inhibition. Proliferation and serum-induced progression of Anx-A1(+/+) cells from G(0)/G(1) into S phase, and the associated expression of extracellular signal-regulated kinase 2 (ERK2), cyclin-dependent kinase 4 (cdk4) and COX 2, is strongly inhibited by dexamethasone, whereas Anx-A1(-/-) cells are refractory to the drug. Loss of the response to dexamethasone in Anx-A1(-/-) cells occurs against a background of no apparent change in glucocorticoid receptor expression or sensitivity to non-steroidal anti-inflammatory drugs. Taken together, these observations suggest strongly that Anx-A1 functions as an inhibitor of signal-transduction pathways that lead to cell proliferation and may help to explain how glucocorticoids regulate these processes.

A Rho-associated kinase mitigates reperfusion-induced change in the shape of cardiac capillary endothelial cells in situ.

OBJECTIVE: We have previously demonstrated that ischaemia and reperfusion of the myocardium alter capillary dimensions and endothelial cell shape and that these changes are likely to be effected by the actomyosin contractile system in endothelial cells. Rho GTPases are involved in the regulation of cytoskeletal re-organization and in cell contraction. Rho-associated kinase regulates the sensitivity of myosin light chain to Ca(2+) in smooth muscle but not in cardiac or skeletal muscle myocytes. This study investigated the role of Rho-associated kinase in endothelial cell shape change induced by cardiac ischaemia and reperfusion. The role of Rho proteins in endothelial cell shape change in situ in the myocardial capillary bed has to date not been investigated. METHODS: Ischaemia and reperfusion were induced in Langendorff perfused rat hearts at constant flow. Electron microscopy and immunofluorescence studies localized the beta Rho-associated kinase isotype in capillary endothelial cells. Whole capillary and luminal cross-section areas, luminal and abluminal membrane lengths were measured to monitor changes in cell dimensions. We used a ROCK inhibitor, Y-27632, to investigate the role of this protein in endothelial cell shape change. RESULTS: ROCK1 localized primarily to intracellular membranes in endothelial cells. Morphometric analysis and a study of capillary lumen resin casts demonstrated that inhibition of the activity of this kinase with Y-27632 ablated the change in shape of endothelial cells induced by ischaemia and reperfusion. CONCLUSION: These results suggest that ROCK1 is involved in cardiac capillary endothelial cell shape change in situ and that targeting the contractile system in this way may be useful in ameliorating reperfusion injury.

Oxidative stress and cardiac microvascular structure in ischemia and reperfusion: the protective effect of antioxidant vitamins.

Reperfusion of the ischemic myocardium results in structural changes in the capillary bed, which may contribute to decreased microcirculatory flow ("no reflow"). This study was designed to correlate the endothelial cell shape changes with both oxidative stress and lipid peroxidation and to evaluate the beneficial potential of Trolox (a hydrophilic analogue of alpha-tocopherol) and ascorbic acid. Isolated buffer-perfused rat hearts were made ischemic for 45 min and then reperfused with 100 microM Trolox and/or 100 microM ascorbic acid. Morphological changes were quantified by measuring capillary cross-sectional areas. Increased myocardial content of oxidized glutathione and its release into the coronary effluent were used as indices of oxidative stress. Myocardial MDA, an end product of lipid peroxidation, was also measured. Luminal membrane blebs and capillary "constriction" in the ischemic groups occurred when there was no change in either glutathione status or MDA concentrations. Reperfusion altered the redox state of the heart sufficiently to induce lipid peroxidation. It also induced endothelial cell swelling and a reduction in luminal area. Ascorbic acid was a more effective antioxidant than Trolox as it significantly reduced both oxidative stress and ultrastructural injury. The combined antioxidant treatment returned both the stress ratio and the capillary measurements to control values. We conclude that endothelial cell swelling correlates with the degree of oxidative stress and that antioxidant vitamins reduce membrane damage by preventing lipid peroxidation.

Ultrastructural and morphometric comparison of retinal and myocardial capillaries following acute ischaemia.

The recovery of any tissue following a period of ischaemia is dependent on a patent microvasculature to restore blood flow. In the ischaemic myocardium, a reduction in capillary cross-sectional dimensions occurs, which is likely to contribute to "no-reflow" injury. Clinical and experimental evidence indicates that the retina is able to tolerate moderate periods of ischaemia without significant loss of function. The aim of the present study is to test the hypothesis that, as an end-arterial system, the retina possesses compensatory processes to maintain a functional microcirculation following acute ischaemia. Thirty minutes of no-flow global ischaemia was induced in isolated hearts of Wistar rats without reperfusion. The retina was also made ischaemic for 30 min using two experimental models: microsphere embolization and anoxic superfusion. Changes in capillary dimensions were assessed by ultrastructural morphometry. Following 30 min of myocardial ischaemia capillaries appeared swollen with a significant reduction in total capillary and luminal cross-sectional area. By contrast, ischaemic retinal capillaries showed minimal morphological changes and no significant alteration in dimensions. We have demonstrated notable differences in the response of retinal and myocardial microvessels to acute ischaemia. It is likely that the maintenance of capillary patency following short periods ischaemia in the retina is part of an adaptive mechanism to protect visual function.

Changes in the actin cytoskeleton of cardiac capillary endothelial cells during ischaemia and reperfusion: the effect of phalloidin on cell shape.

A reduction in capillary dimensions has been demonstrated in postischaemic reperfusion in the heart. The aim of this study was to demonstrate that in ischaemia and ischaemia followed by reperfusion, the change in shape of the constituent endothelial cells can be inhibited by phalloidin which stabilises the actin microfilament system. Isolated, perfused rat hearts were made globally ischaemic both with and without reperfusion and in the presence or absence of phalloidin. Changes in ischaemic endothelial cell dimensions were quantified by measuring whole capillary and luminal cross-sectional areas, abluminal and luminal membrane lengths. The distribution of beta-actin within the endothelial cells was determined by immunocytochemistry. In control hearts, beta-actin is distributed throughout the endothelium with a slight increase towards the luminal membrane. In ischaemia, this was more marked and other patterns of actin distribution were also observed. After reperfusion, a 'double ring' of actin could be distinguished. With phalloidin, the actin staining was more regular and the ring pattern was not observed. Morphometry showed that phalloidin was more effective in reducing endothelial cell shape change after reperfusion than after ischaemia alone. We conclude that endothelial cell shape change on reperfusion can be modified by agents which target the contractile proteins.