Quantification of microbial risks to human health caused by waterborne viruses and bacteria in an urban slum.

AIMS: To determine the magnitude of microbial risks from waterborne viruses and bacteria in Bwaise III in Kampala (Uganda), a typical slum in Sub-Saharan Africa. METHODS AND RESULTS: A quantitative microbial risk assessment (QMRA) was carried out to determine the magnitude of microbial risks from waterborne pathogens through various exposure pathways in Bwaise III in Kampala (Uganda). This was based on the concentration of Escherichia coli O157:H7, Salmonella spp., rotavirus (RV) and human adenoviruses F and G (HAdV) in spring water, tap water, surface water, grey water and contaminated soil samples. The total disease burden was 680 disability-adjusted life years (DALYs) per 1000 persons per year. The highest disease burden contribution was caused by exposure to surface water open drainage channels (39%) followed by exposure to grey water in tertiary drains (24%), storage containers (22%), unprotected springs (8%), contaminated soil (7%) and tap water (0.02%). The highest percentage of the mean estimated infections was caused by E. coli O157:H7 (41%) followed by HAdV (32%), RV (20%) and Salmonella spp. (7%). In addition, the highest infection risk was 1 caused by HAdV in surface water at the slum outlet, while the lowest infection risk was 2.71 × 10(-6) caused by E. coli O157:H7 in tap water. CONCLUSIONS: The results show that the slum environment is polluted, and the disease burden from each of the exposure routes in Bwaise III slum, with the exception of tap water, was much higher than the WHO reference level of tolerable risk of 1 × 10(-6) DALYs per person per year. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this study provide guidance to governments, local authorities and nongovernment organizations in making decisions on measures to reduce infection risk and the disease burden by 10(2) to 10(5) depending on the source of exposure to achieve the desired health impacts. The infection risk may be reduced by sustainable management of human excreta and grey water, coupled with risk communication during hygiene awareness campaigns at household and community level. The data also provide a basis to make strategic investments to improve sanitary conditions in urban slums.

Genomic copy concentrations of selected waterborne viruses in a slum environment in Kampala, Uganda.

The presence of viruses in a slum environment where sanitation is poor is a major concern. However, little is known of their occurrence and genomic copy concentration in the slum environment. The main objective of this study was to determine the genomic copy concentrations of human adenoviruses F and G, Rotavirus (RV), Hepatitis A virus (HAV), Hepatitis E virus (HEV) and human adenovirus species A,C,D,E, and F (HAdV-ACDEF) in Bwaise III, a typical slum in Kampala, Uganda. Forty-one samples from surface water, grey water and ground water were collected from 30 sampling locations. The virus particles were recovered by glass wool filtration with elution using beef extract. DNA and RNA viruses were detected by the real time quantitative polymerase chain reaction (qPCR) and the reverse transcription-qPCR (RT-qPCR), respectively. HAdV-F and G were detected in 70.7% of the samples with concentrations up to 2.65 × 10(1) genomic copies per mL (gc mL(-1)). RV and HAV were detected in 60.9% and 17.1% of the samples, respectively. The maximum concentration of RV was 1.87 × 10(2)gc mL(-1). In addition, 78% of the samples tested positive for the HAdV-ACDEF, but all samples tested negative for HEV. These new data are essential for quantitative microbial risk assessment, and for understanding the effects of environmental pollution in slums.

Transport of Escherichia coli strains isolated from natural spring water.

We present a new methodology to scale up bacteria transport experiments carried out in the laboratory to practical field situations. The key component of the methodology is to characterize bacteria transport not by a constant sticking efficiency, but by a range of sticking efficiency values determined from laboratory column experiments. In this study, initially, we harvested six Escherichia coli strains from springs in Kampala, the capital of Uganda, and then we carried out a number of experiments with 1.5m high columns of quartz sand with various sampling ports in order to determine the fraction of bacteria as a function of sticking efficiency. Furthermore, we developed a simple mathematical formulation, based on the steady-state analytical solution for the transport of mass in the subsurface, to arrive at bacteria concentrations as a function of transport distance. The results of the quartz sand column experiments indicated that the fractional bacteria mass and sticking efficiency of most of the strains we harvested could be adequately described by a power law. When applying the power distributions to the field situation in Kampala, we found that the transport distance required to reduce bacteria concentrations with five log units ranged from 1.5 to 23m, and this was up to three times more than when using a constant sticking efficiency. The methodology we describe is simple, can be carried out in a spreadsheet, and in addition to parameters describing transport, like pore water flow velocity and dispersion, only two constants are required, which define the relation between sticking efficiency and percentage of bacteria mass.

Sustainable sanitation technology options for urban slums.

Poor sanitation in urban slums results in increased prevalence of diseases and pollution of the environment. Excreta, grey water and solid wastes are the major contributors to the pollution load into the slum environment and pose a risk to public health. The high rates of urbanization and population growth, poor accessibility and lack of legal status in urban slums make it difficult to improve their level of sanitation. New approaches may help to achieve the sanitation target of the Millennium Development Goal (MDG) 7; ensuring environmental sustainability. This paper reviews the characteristics of waste streams and the potential treatment processes and technologies that can be adopted and applied in urban slums in a sustainable way. Resource recovery oriented technologies minimise health risks and negative environmental impacts. In particular, there has been increasing recognition of the potential of anaerobic co-digestion for treatment of excreta and organic solid waste for energy recovery as an alternative to composting. Soil and sand filters have also been found suitable for removal of organic matter, pathogens, nutrients and micro-pollutants from grey water.

Transport of Escherichia coli in 25 m quartz sand columns.

To help improve the prediction of bacteria travel distances in aquifers laboratory experiments were conducted to measure the distant dependent sticking efficiencies of two low attaching Escherichia coli strains (UCFL-94 and UCFL-131). The experimental set up consisted of a 25 m long helical column with a diameter of 3.2 cm packed with 99.1% pure-quartz sand saturated with a solution of magnesium sulfate and calcium chloride. Bacteria mass breakthrough at sampling distances ranging from 6 to 25.65 m were observed to quantify bacteria attachment over total transport distances (α(L)) and sticking efficiencies at large intra-column segments (α(i)) (>5m). Fractions of cells retained (F(i)) in a column segment as a function of α(i) were fitted with a power-law distribution from which the minimum sticking efficiency defined as the sticking efficiency of 0.001% bacteria fraction of the total input mass retained that results in a 5 log removal were extrapolated. Low values of α(L) in the order 10(-4) and 10(-3) were obtained for UCFL-94 and UCFL-131 respectively, while α(i)-values ranged between 10(-6) to 10(-3) for UCFL-94 and 10(-5) to 10(-4) for UCFL-131. In addition, both α(L) and α(i) reduced with increasing transport distance, and high coefficients of determination (0.99) were obtained for power-law distributions ofα(i) for the two strains. Minimum sticking efficiencies extrapolated were 10(-7) and 10(-8) for UCFL-94 and UCFL-131, respectively. Fractions of cells exiting the column were 0.19 and 0.87 for UCFL-94 and UCL-131, respectively. We concluded that environmentally realistic sticking efficiency values in the order of 10(-4) and 10(-3) and much lower sticking efficiencies in the order 10(-5) are measurable in the laboratory, Also power-law distributions in sticking efficiencies commonly observed for limited intra-column distances (<2m) are applicable at large transport distances(>6m) in columns packed with quartz grains. High fractions of bacteria populations may possess the so-called minimum sticking efficiency, thus expressing their ability to be transported over distances longer than what might be predicted using measured sticking efficiencies from experiments with both short (<1m) and long columns (>25 m). Also variable values of sticking efficiencies within and among the strains show heterogeneities possibly due to variations in cell surface characteristics of the strains. The low sticking efficiency values measured express the importance of the long columns used in the experiments and the lower values of extrapolated minimum sticking efficiencies makes the method a valuable tool in delineating protection areas in real-world scenarios.

Determining minimum sticking efficiencies of six environmental Escherichia coli isolates.

In health impact assessments, the sticking efficiency of a bacteria or virus population largely determines the transported distance of that biocolloid population, and hence, the potential health impact. However, at the same time, one of the most difficult parameters to estimate is the lower value of the sticking efficiency that should be used in calculating the health impact. In this paper, we introduce the concept of the minimum sticking efficiency (alpha(i)) value of a bacteria population, including a method to determine the minimum sticking efficiency. Thereto, sticking efficiency distributions of 6 environmentally isolated Escherichia coli strains were determined by carrying out laboratory column experiments over a transport distance of about 5m. Experiments were conducted in de-mineralized (DI) water and in artificial groundwater (AGW). Sticking efficiencies were calculated for column segments (at varying distances from top of column) and fractions of total bacteria mass input in each segment were estimated by mass balance. The sticking efficiencies were highest close to the top of the column, near the point of bacteria mass input (0.103-0.352 in DI, and 1.034-9.470 for AGW) and reduced with distance with the lowest alpha(i) values (10(-5)-0.06 in DI and 0.006-0.283 in AGW) determined at the two most distant column segments (between 2.33 and 4.83 m from the top of the column). Power-law distributions best described the relationship between fraction of cells retained, F(i), and alpha(i). The minimum sticking efficiency was defined as the sticking efficiency belonging to a retained bacteria fraction of 0.001% of the original bacteria mass (total number of cells) flowing into the column (F=10(-5)), and coinciding with a 99.999% reduction of the original bacteria mass, and minimum sticking efficiencies were extrapolated from the fitted power-law distributions. In the DI experiments, minimum sticking efficiency values ranged from as low as 10(-9) (for E. coli strain UCFL-94) to 10(-2) (for E. coli strain UCFL-348); in the AGW experiments, minimum sticking efficiency values ranged from 10(-6) (for strain UCFL-94) to > or =1(for strain UCFL-348). We concluded that in quantifying health impacts of biocolloids traveling in aquifers, the concept of the minimum sticking efficiencies, and the percentage of individual biocolloids of a total population having such low sticking efficiency, together with an inactivation rate coefficient, can serve as a useful tool to determine the maximum transported distance as a worst case scenario, and, hence, the potential health impact.

The effect of surface characteristics on the transport of multiple Escherichia coli isolates in large scale columns of quartz sand.

Bacteria properties play an important role in the transport of bacteria in groundwater, but their role, especially for longer transport distances (>0.5 m) has not been studied. Thereto, we studied the effects of cell surface hydrophobicity, outer surface potential (OSP), cell sphericity, motility, and Ag43 protein expression on the outer cell surface for a number of E. coli strains, obtained from the environment on their transport behavior in columns of saturated quartz sand of 5 m height in two solutions: demineralized (DI) water and artificial groundwater (AGW). In DI water, sticking efficiencies ranged between 0.1 and 0.4 at the column inlet, and then decreased with transport distance to 0.02-0.2. In AGW, sticking efficiencies were on average 1log-unit higher than those in DI (water). Bacteria motility and Ag43 expression affected attachment with a (high) statistical significance. In contrast, hydrophobicity, OSP and cell sphericity did not significantly correlate with sticking efficiency. However, for transport distances more than 0.33 m, the correlation between sticking efficiency, Ag43 expression, and motility became insignificant. We concluded that Ag43 and motility played an important role in E. coli attachment to quartz grain surfaces, and that the transport distance dependent sticking efficiency reductions were caused by motility and Ag43 expression variations within a population. The implication of our findings is that less motile bacteria with little or no Ag43 expression may travel longer distances once they enter groundwater environments. In future studies, the possible effect of bacteria surface structures, like fimbriae, pili and surface proteins on bacteria attachment need to be considered more systematically in order to arrive at more meaningful inter-population comparisons of the transport behavior of E. coli strains in aquifers.

Transport of Escherichia coli and solutes during waste water infiltration in an urban alluvial aquifer.

Recharge of waste water in an unconsolidated poorly sorted alluvial aquifer is a complex process, both physically and hydrochemically. The aim of this paper is to analyse and conceptualise vertical transport mechanisms taking place in an urban area of extensive wastewater infiltration by analysing and combining the water balance, the microbial (Escherichia coli) mass balance, and the mass balance for dissolved solutes. For this, data on sediment characteristics (grain size, organic carbon, reactive iron, and calcite), groundwater levels, and concentrations of E. coli in groundwater and waste water were collected. In the laboratory, data on E. coli decay rate coefficients, and on bacteria retention characteristics of the sediment were collected via column experiments. The results indicated that shallow groundwater, at depths of 50 m below the surface, was contaminated with E. coli concentrations as high as 10(6) CFU/100 mL. In general, E. coli concentrations decreased only 3 log units from the point of infiltration to shallow groundwater. Concentrations were lower at greater depths in the aquifer. In laboratory columns of disturbed sediments, bacteria removal was 2-5 log units/0.5 cm column sediment. Because of the relatively high E. coli concentrations in the shallow aquifer, transport had likely taken place via a connected network of pores with a diameter large enough to allow bacterial transport instead of via the sediment matrix, which was inaccessible for bacteria, as was clear from the column experiments. The decay rate coefficient was determined from laboratory microcosms to be 0.15 d(-1). Assuming that decay in the aquifer was similar to decay in the laboratory, then the pore water flow velocity between the point of infiltration and shallow groundwater, coinciding with a concentration decrease of 3 log units, was 0.38 m/d, and therefore, transport in this connected network of pores was fast. According to the water balance of the alluvial aquifer, determined from transient groundwater modelling, groundwater flow in the aquifer was mainly in vertical downward direction, and therefore, the mass balance for dissolved solutes was simulated using a 1D transport model of a 200 m column of the Quaternary Alluvium aquifer. The model, constructed with PHREEQC, included dual porosity, and was able to adequately simulate removal of E. coli, cation-exchange, and nitrification. The added value of the use of E. coli in this study was the recognition of relatively fast transport velocities occurring in the aquifer, and the necessity to use the dual porosity concept to investigate vertical transport mechanisms. Therefore, in general and if possible, microbial mass balances should be considered more systematically as an integral part of transport studies.

Effect of goethite coating and humic acid on the transport of bacteriophage PRD1 in columns of saturated sand.

The transport of bacteriophage PRD1, a model virus, was studied in columns containing sediment mixtures of quartz sand with goethite-coated sand and using various solutions consisting of monovalent and divalent salts and humic acid (HA). Without HA and in the absence of sand, the inactivation rate of PRD1 was found to be as low as 0.014 day(-1) (at 5+/-3 degrees C), but in the presence of HA it was much lower (0.0009 day(-1)), indicating that HA helps PRD1 to survive. When the fraction of goethite in the sediment was increased, the removal of PRD1 also increased. However, in the presence of HA, C/C0 values of PRD1 increased by as much as 5 log units, thereby almost completely eliminating the effect of addition of goethite. The sticking efficiency was not linearly dependent on the amount of goethite added to the quartz sand; this is apparently due to surface charge heterogeneity of PRD1. Our results imply that, in the presence of dissolved organic matter (DOM), viruses can be transported for long distances thanks to two effects: attachment is poor because DOM has occupied favourable sites for attachment and inactivation of virus may have decreased. This conclusion justifies making conservative assumptions about the attachment of viruses when calculating protection zones for groundwater wells.

Evaluation of data from the literature on the transport and survival of Escherichia coli and thermotolerant coliforms in aquifers under saturated conditions.

Escherichia coli and thermotolerant coliforms are of major importance as indicators of fecal contamination of water. Due to its negative surface charge and relatively low die-off or inactivation rate coefficient, E. coli is able to travel long distances underground and is therefore also a useful indicator of fecal contamination of groundwater. In this review, the major processes known to determine the underground transport of E. coli (attachment, straining and inactivation) are evaluated. The single collector contact efficiency (SCCE), eta0, one of two parameters commonly used to assess the importance of attachment, can be quantified for E. coli using classical colloid filtration theory. The sticking efficiency, alpha, the second parameter frequently used in determining attachment, varies widely (from 0.003 to almost 1) and mainly depends on charge differences between the surface of the collector and E. coli. Straining can be quantified from geometrical considerations; it is proposed to employ a so-called straining correction parameter, alpha(str). Sticking efficiencies determined from field experiments were lower than those determined under laboratory conditions. We hypothesize that this is due to preferential flow mechanisms, E. coli population heterogeneity, and/or the presence of organic and inorganic compounds in wastewater possibly affecting bacterial attachment characteristics. Of equal importance is the inactivation or die-off of E. coli that is affected by factors like type of bacterial strain, temperature, predation, antagonism, light, soil type, pH, toxic substances, and dissolved oxygen. Modeling transport of E. coli can be separated into three steps: (1) attachment rate coefficients and straining rate coefficients can be calculated from Darcy flow velocity fields or pore water flow velocity fields, calculated SCCE fields, realistic sticking efficiency values and straining correction parameters, (2) together with the inactivation rate coefficient, total rate coefficient fields can be generated, and (3) used as input for modeling the transport of E. coli in existing contaminant transport codes. Areas of future research are manifold and include the effects of typical wastewater characteristics, including high concentrations of organic compounds, on the transport of E. coli and thermotolerant coliforms, and the upscaling of experiments to represent typical field conditions, possibly including preferential flow mechanisms and the aspect of population heterogeneity of E. coli.

Transport of E. coli in columns of geochemically heterogeneous sediment.

To elucidate the parameters determining the transport of Escherichia coli in aquifers, the attachment of E. coli in low concentrations to column sediments was investigated. The sediments comprised 0.18-0.50mm quartz sand, grains coated with goethite, calcite grains or grains of activated carbon (AC), in varying fractions (lambda=0, 0.05, 0.1, 0.2, 0.4, 0.7, 1.0) and all of similar diameter to the quartz sand. The weighted sum of favourable and unfavourable sticking efficiencies (alpha(total)) showed that upon increasing the fraction of favourable mineral grains (lambda) there was an initial rapid increase, which then slowed down. This was most pronounced in the AC experiments, followed by the calcite experiments and then the goethite experiments. We ascribe this non-linear relation to surface charge and hydrophobic heterogeneity of the E. coli population.

Determining straining of Escherichia coli from breakthrough curves.

Though coliform bacteria are used world wide as an indication of faecal pollution, the parameters determining the transport of Escherichia coli in aquifers are relatively unknown, especially for the period after the clean bed collision phase brought about by prolonged infiltration of waste water. In this research, the breakthrough curves of E. coli after total flushing of 50-200 pore volumes were studied for various influent concentrations in various sediments at different pore water flow velocities. The results indicated that straining in Dead End Pores (DEPs) was an important process that dominated bacteria breakthrough in fine-grained sediment (0.06-0.2 mm). The filling of the DEP space with bacteria took 5-65 pore volumes and was dependent on concentration. Column breakthrough curves were modelled and from this the DEP volumes were determined. These volumes (0.21-0.35% of total column volume) corresponded well with values calculated with a formula based on purely geometrical considerations and also with values calculated with a pore size density function. For this function the so-called Van Genuchten parameters of the sediments used in the experiments were determined. The results indicate that straining might be a dominant process affecting colloid transport in the natural environment and therefore it is concluded that proper knowledge of the pore size distribution is crucial to an understanding of the retention of bacteria.