Liming as an advanced treatment for sludge sanitisation: helminth eggs elimination--Ascaris eggs as model

The presence of helminth eggs (Ascaris eggs) in sewage sludge may constitute a sanitary risk when used as agricultural fertiliser. Sanitisation of sewage sludge can be achieved by treatment with quick lime, a process that destroys sludge pathogens in two ways: pH increase and temperature rise. Among the pathogens of epidemiological relevance, Ascaris eggs are the most resistant to liming, and, hence, may serve as indicators of hygienic quality of biosolids. This research aims at defining, between 50 degrees C and 60 degrees C, the time required in the case of limed sludge to obtain a product with a negligible level of viable Ascaris eggs. To achieve this objective, investigations on inactivation kinetics of Ascaris eggs were conducted in the following products: contaminated milk of lime; naturally contaminated sludge treated with slaked lime and heat; naturally contaminated sludge treated with quick lime; and sludge treated at full scale with quick lime. For the inactivation kinetics where a negligible level of Ascaris eggs was reached, the inactivation threshold was determined. Depending on the experimental situation, the inactivation threshold period was found to fluctuate between 5 and 75 min at 55 degrees C and between 1 and 8 min at 60 degrees C.     

Viability of six species of larval and non-larval helminth eggs for different conditions of temperature, pH and dryness

Helminth eggs are the most difficult biological parasites to inactivate in wastewater and sludge. In developing countries, in particular, they are present in high concentrations and are the cause of many diseases that impact seriously on the human population. The process conditions for affordable inactivation are very variable, leading to different inactivation efficiencies. Temperature, dryness, pH and the developmental stage of the eggs must be taken into consideration to achieve complete inactivation. The objective of this research was to study the inactivation of six species of larval and non-larval helminth eggs of medical importance in developing countries under controlled conditions of temperature, pH, dryness and contact time. Results showed considerable differences in inactivation conditions among helminth eggs and a high level of resistance was confirmed for the eggs of Ascaris lumbricoides and Ascaris suum. The appropriate conditions for inactivation of all types of eggs were found by applying combinations of pH, temperature and dryness. At 45 °C it was possible to inactivate all species with a pH of 5.3 and 90% dryness within 6 days. If alkalization was applied, a pH of 12.7 was sufficient over 19 days at the same conditions of dryness and temperature. From these results it is proposed that both Ascaris spp. and Taenia solium may be used as indicators of biological contamination in wastewater and sludge.     

The potential for self-sanitisation of faecal sludge by intrinsic ammonia

Faecal sludge has the potential to be used as a sustainable fertiliser in agriculture, but the sludge must be sanitised due to its content of pathogenic microorganisms. The intrinsic ammonia from the urine may be sufficient for sanitisation of the sludge if it is not too diluted by flush water or lost by ventilation. To evaluate the potential for this sanitisation method, inactivation of Enterococcus faecalis, Salmonella typhimurium and Ascaris suum eggs during treatment were assessed. The inactivation was studied at different storage temperatures (10–28 °C) and in several sludge mixes with different contents of urine, faeces and flush water, and with ammonia concentrations from 40 to 400 mM. All pathogens were inactivated by the ammonia, and ascaris eggs were the most persistent. Lower flush water volume and higher urine content favoured inactivation, mainly due to increased uncharged ammonia (NH₃) concentration. The lag phase in ascaris inactivation was shortened by increasing temperature and NH₃ concentration, while post-lag phase inactivation was not influenced by NH₃ concentration. Faecal sludge can be sanitised by airtight storage without the use of additives when flush water volumes are sufficiently low. For temperatures of 23–28 °C, a 3 log reduction of ascaris egg viability can be achieved within 1–6 months depending on ammonia concentration and temperature.     

Inactivation of bacteria and viruses in human urine depending on temperature and dilution rate

Source separation and reuse of human urine can decrease the environmental pollution of recipient waters and reduce the need for artificial mineral fertilisers. However, the reuse of urine introduces another pathogen transmission route that needs to be managed. The inactivation of enteric pathogens and model organisms (Salmonella enterica subspecies 1 serovar Typhimurium (S. typhimurium), Enterococcus faecalis, bacteriophages S. typhimurium 28B, MS2 and Phix 174) by urine storage was studied at dilutions (urine:water) 1:0, 1:1 and 1:3 at temperatures 4, 14, 24 and 34 degrees C. A threshold concentration of ammonia was found at approximately 40mMNH(3) (e.g. 2.1gNH(3)-NL(-1) and pH 8.9 at 24 degrees C), below which all studied organisms, except Salmonella, persisted considerably longer irrespective of treatment temperature, showing that urine dilution rate is of great importance for pathogen inactivation. For Salmonella spp. no threshold level was found in these studies (15mMNH(3) lowest concentration studied). At temperatures below 20 degrees C, bacteriophage reduction was very slow. Therefore, urine stored at temperatures below 20 degrees C carries a high risk of containing viable viruses. The study indicated that the current recommended storage time for urine of 6 months at 20 degrees C or higher is safe for unrestricted use and could probably be shortened, especially for undiluted urine.     

Sludge accumulation, characteristics, and pathogen inactivation in four primary waste stabilization ponds in central Mexico

To support the development of safe and feasible sludge management strategies, the accumulation rates of sludge and its characteristics were studied in four primary wastewater stabilization ponds (WSPs) in central Mexico (three facultative and one anaerobic). The accumulation rates and distribution of sludge were determined by measuring the thickness of the sludge layer at 8-40 locations throughout each pond. The average, per capita sludge accumulation rates ranged from 0.021 to 0.036m(3)/person/yr. In the anaerobic pond the sludge distribution was uniform throughout the pond, whereas in the three facultative ponds most of the sludge accumulated directly in front of the inlet. To measure the horizontal and vertical variation in the sludge characteristics, sludge cores were collected from 3 to 7 locations in three of the ponds. Each core was divided into 4 sub-samples in which various physical, chemical, and microbiological parameters were measured. In addition, the inactivation of several pathogen indicator organisms was studied in a batch of sludge for 7 months. Based on the microbiological results, it is concluded that reasonable estimates of the inactivation of fecal coliform bacteria, fecal enterococci, F+ coliphage, somatic coliphage, and Ascaris eggs in WSP sludge in central Mexico can be made using first-order rate constants of 0.1, 0.1, 0.01, 0.001, and 0.001d(-1), respectively. From the observed changes in the concentrations of total solids and the volatile to fixed solids ratio, empirical equations were developed to describe anaerobic degradation and compression, which are the two most important processes affecting the volume of sludge after its deposition.     

Inactivation of Bacillus subtilis spores and formation of bromate during ozonation

Inactivation of B. subtilis spores with ozone was investigated to assess the effect of pH and temperature, to compare the kinetics to those for the inactivation of C. parvum oocysts, to investigate bromate formation under 2-log inactivation conditions, and to assess the need for bromate control strategies. The rate of B. subtilis inactivation with ozone was independent of pH, decreased with temperature (activation energy of 42,100 Jmol(-1)), and was consistent with the CT concept. B. subtilis was found to be a good indicator for C. parvum at 20-30 degrees C, but at lower temperatures B. subtilis was inactivated more readily than C. parvum. Bromate formation increased as both pH and temperature increased. For water with an initial bromide concentration of 33 microgl(-1), achieving 2-logs of inactivation, without exceeding the 100 microg l(-1) bromate standard, was most difficult at 30 degrees C for B. subtilis and at midrange temperatures (10-20 degrees C) for C. partum. pH depression and ammonia addition were found to reduce bromate formation without affecting B. subtilis inactivation, and may be necessary for waters containing more than 50 microgl(-1) bromide.     

Helminth eggs inactivation efficiency by faecal sludge dewatering and co-composting in tropical climates

This study investigates helminth eggs removal and inactivation efficiency in a treatment process combining faecal sludge (FS) dewatering and subsequent co-composting with organic solid waste as a function of windrow turning frequency. Fresh public toilet sludge and septage mixed at a 1:2 ratio were dewatered on a drying bed. Biosolids with initial loads of 25-83 helminth eggs/g total solids (TS) were mixed with solid waste as bulking material for co-composting at a 1:2 volume ratio. Two replicate sets of compost heaps were mounted in parallel and turned at different frequencies during the active composting period: (i) once every 3 days and (ii) once every 10 days. Turning frequency had no effect on helminth eggs removal efficiency. In both setups, helminth eggs were reduced to <1 viable egg/g TS, thereby complying with the WHO guidelines 2006 for the safe reuse of FS.     

Inactivation kinetics of adenovirus serotype 2 with monochloramine

The effect of pH (6-10), temperature (10-30 degrees C), disinfectant concentration (1-11mg/l as Cl(2)), and ammonia nitrogen-to-chlorine molar ratio (1.3-52) on the inactivation kinetics of adenovirus serotype 2 with monochloramine was investigated by performing batch-reactor experiments with synthetic 0.01M buffer (phosphate or borate) solutions. The inactivation kinetics was independent of monochloramine concentration and ammonia nitrogen-to-chlorine molar ratio but had strong pH dependence, with the rate of inactivation decreasing with increasing pH. The kinetics at pH 6 and 8 were consistent with pseudo-first-order kinetics, while curves at pH 10 were characterized by a lag phase followed by a pseudo-first-order phase. The rate of inactivation increased with increasing temperature-activation energies of 56.5kJ/mole (pH 8) and 72.6kJ/mole (pH 10). The results obtained in this study revealed that monochloramine disinfection might not generally provide adequate control of adenoviruses in drinking water at high pH and low temperature.     

Influence of biotic and abiotic factors on human pathogens in a finished compost

The role of indigenous microflora of a finished compost, defined NK12, on the growth suppression of pathogens under different moisture and temperature storages was investigated. Total count of mesophilic and thermophilic bacteria was evaluated by the most probable number method and growth of seeded Salmonella arizonae 3924 serogroup B and enteropathogenic Escherichia coli 84 M in NK12 at different moisture temperature conditions was monitored. Results on sterile and non-sterile NK12 were compared. In all tested experimental conditions, the NK12 indigenous microflora was stable and biologically active. S. arizonae 3924 and E. coli 84 M grew rapidly in sterilized NK12 at different moistures and storage temperatures, and their growth was suppressed in non-sterilized NK12. Pathogens inactivation was lower when compost was stored at 40% and 80% humidity and at 37 degrees C. Our results show that the major role in the pathogens suppression was played by the indigenous microflora of the finished compost, although physical factors too influenced the growth phenomenon.     

Factors influencing inactivation of Klebsiella pneumoniae by chlorine and chloramine

Inactivation of Klebsiella pneumoniae cultures by chlorine and chloramine was evaluated under different growth conditions by varying nutrient media dilution, concentrations of essential inorganic nutrients (FeCl3, MgSO4, phosphate, and ammonium salts), and temperature. All inactivation assays were performed at room temperature (22-23 degrees C) and near neutral pH (7.2-7.5). C*T(99.9) values for chlorine increased >20-fold and for chloramine increased 2.6-fold when cells were grown in 100-fold diluted nutrient broth (2NB) solutions (final TOC of 35-40 mg/L). Background levels of Mg: 6.75 x 10(-2) mM and Fe: 3.58 x 10(-5) mM or high levels of FeCl3 (0.01 mM) and MgSO4 (1 mM) during growth resulted in the highest resistances to chlorine with C*T(99.9) values of 13.06 (+/-0.91) and 13.78 (+/-1.97) mg-min/L, respectively. Addition of low levels of FeCl3 (0.001 mM) and MgSO4 (0.1 mM) to K. pneumoniae cultures during growth resulted in the lowest bacterial resistances to inactivation; C*T(99.9) values ranged from 0.28 (+/-0.06) to 1.88 (+/-0.53)mg-min/L in these cultures. Increase in growth temperature from 22.5 degrees C to 35 degrees C for unamended 2NB cultures resulted in a 42-fold decrease in C*T(99.9) values for chlorine. A similar change in temperature resulted in no significant change in C*T(99.9) values for chloramine. These results indicate that inactivation of K. pneumoniae cultures by chlorine was highly sensitive to changes in growth conditions unlike inactivation by chloramine.     

Cryptosporidium parvum oocyst inactivation in field soil and its relation to soil characteristics: analyses using the geographic information systems

The need exists to understand the environmental parameters that affect inactivation of Cryptosporidium parvum oocysts in soil under field conditions. The inactivation of C. parvum oocysts placed in the natural environment was studied at a dairy farm in western New York State, USA. Seventy sampling points were arranged in a grid with points 150 m apart using the Geographic Information System. The sampling points were distributed among three distinct areas: woodland, corn field and pasture. Purified oocysts were inoculated into chambers filled with soil from each sampling point, and buried in the surface of each respective sampling point. To compare C. parvum oocyst survival with another organism known to survive environmental stresses, Ascaris suum eggs were also placed in soil contained in chambers and buried at the same sampling points as the oocysts. As controls oocysts and eggs in distilled water were also placed at each sampling point. Oocyst and egg viability, soil pH and percent gravimetric water content were measured at all sampling points at 0, 60 and 120 day sampling periods. Soil organic content was determined for each sampling point. At 120 days after placement, mean viability of C. parvum oocysts was 10% although at a few sampling points, 30% of oocysts were still potentially infective; whereas 90% of A. suum eggs were viable at all sampling points. Statistically significant differences were not observed among the three different sampling areas, and no statistically significant predictors were found by regression analysis. Results exemplified the heterogeneity of soil parameters and oocyst viability across a landscape; such results make predictive models for C. parvum inactivation problematical. The long-term survival of C. parvum oocysts in soil under field conditions, as this study demonstrated, emphasizes their potential as a risk to contaminate surface waters.     

Hydrolysis and acidification of waste activated sludge at different pHs

The effect of pH from 4.0 to 11.0 on the hydrolysis and acidification of waste activated sludge (WAS) was investigated. Experimental results showed that at room temperature the sludge hydrolysis was in the following order: alkaline>acidic>(neutral and blank test), and between pH 6.0 and 11.0 the sludge hydrolysis increased with pH. The three main components, soluble protein, carbohydrate and volatile fatty acids (VFAs) in the hydrolytic product were analyzed. It was observed that both the soluble protein and carbohydrate increased with pH in the pH range 7.0-11.0, but also increased to a smaller extent with pH from 7.0 to 4.0. The VFAs concentration was also affected by pH. Under alkaline conditions, the VFAs production was significantly higher than under other conditions. The concentration of VFAs on the 8th day of fermentation at pH 4.0, 7.0 and 10.0 was, respectively, 354.49, 842.00 and 2708.02 mg/L, while VFAs in the blank test was only 633.59 mg/L. The VFAs consisted of acetic, propionic, iso-butyric, n-butyric, iso-valeric and n-valeric acids, but acetic, propionic and iso-valeric were the three main products. Also, the release of soluble phosphorus and ammonia and the production of methane was studied during WAS fermentation at different pHs.     

Optimization of the hydrolytic-acidogenic anaerobic digestion stage (55 degrees C) of sewage sludge: influence of pH and solid content

In conventional single-stage anaerobic digestion processes, hydrolysis is regarded as the rate-limiting step in the degradation of complex organic compounds, such as sewage sludge. Two-stage systems have been proposed to enhance this process. However, so far it is not clear which are the best conditions for a two-stage anaerobic digestion process of sewage sludge, in terms of temperature and hydraulic retention time of each stage. The aim of this work was to determine the optimal conditions for the hydrolytic-acidogenic stage treating real sludge with a high concentration of total solids (40-50gL(-1)) and volatile solids (25-30gL(-1)), named high concentration sludge. The variables considered for this first stage were: hydraulic retention time (1-4 days) and temperature (55 and 65 degrees C). Maximum volatile fatty acids generation was obtained at 4 days and 3 days hydraulic retention time for 55 degrees C and 65 degrees C, respectively. Consequently, 4 days hydraulic retention time and temperature of 55 degrees C were set as the working conditions for the hydrolytic-acidogenic stage treating high concentration sludge. The results obtained when operating with high concentration sludge were compared with a low concentration sludge consisting of 17-28gL(-1) total solids and 13-21gL(-1) volatile solids. The effect of decreasing the influent sludge pH, when working at the optimal conditions established, was also evaluated.     

Synergistic inactivation of Cryptosporidium parvum using ozone followed by free chlorine in natural water

The synergistic effect of sequential exposure to ozone followed by free chlorine on inactivation of Cryptosporidium parvum oocysts suspended in natural waters was studied in bench-scale batch reactors. Animal infectivity using neonatal CD-1 mice was used to measure oocyst inactivation. The synergistic effect measured in two alkaline (pH 8.1) natural waters was statistically significant but was considerably smaller than previously reported in buffered de-ionized water at pH 6.0. Temperature, ozone primary treatment level, and water type did not have measurable impacts on the synergistic effect. Efforts to increase the synergistic effect by reducing the pH from 8 to 6 by acid addition were unsuccessful. In the two low alkalinity (pH 6.0) natural waters tested, the measured synergistic effect was greater than in the alkaline waters, but was still less than that measured previously in buffered de-ionized water. It was concluded that the synergistic effect reduction in the natural waters tested was due in part to alkalinity and in part to other unidentified water quality characteristics. Sequential treatment with ozone followed by free chlorine may only be a feasible strategy for achieving synergistic C. parvum inactivation credit for water treatment facilities with natural waters having a low pH (near 6.0).     

Natural freezing as a wastewater treatment method: E. coli inactivation capacity

Inactivation capacity of E. coli (strain ATCC 15597) in water by natural freezing was examined via two freezing methods: spray freezing and freezing in a freezer. The effect of freezing temperature (-5, -15 and -35 degrees C), storage time, freeze-thaw cycles on the survival of the test organism were investigated. In addition, the number of cells injured by the freezing process was also examined by using different growth media. The bacteria frozen at the warmer temperature (-5 degrees C) was most sensitive to storage and freeze-thaw cycles as compared to those frozen at -15 and -35 degrees C. In general, greater inactivation efficiencies were achieved under longer storage time and warmer freezing temperature conditions. Freezing and thawing caused cell injury. More cells were injured when frozen at -15 degrees C. The percentage of cells injured decreased as freeze-thaw cycles increased. The spray-freezing process was found more effective in killing the cells. On average, the log reduction rate for the spray ice with two-day storage time was about 4 log units higher than those without any storage after freezing. The results indicated that the natural freezing processes are not only cost-effective techniques for chemical and physical contaminant removal from wastewater or enhancing sludge dewaterability in cold regions but also effective in reducing E. coli concentration.     

Inactivation of Bacillus subtilis spores with ozone and monochloramine

The inactivation kinetics of Bacillus subtilis spores with ozone and monochloramine was characterized by a lag phase followed by a pseudo-first-order rate of inactivation. The lag phase decreased and the post-lag phase rate constant increased with increasing temperature within the range investigated (1-30 degrees C for ozone, 1-20 degrees C for monochloramine). The corresponding activation energies were 46820 J/mol for ozone and 79640 J/mol for monochloramine. The CT concept was found to be valid within the concentration range investigated of 0.44-4.8 mg/l for ozone, and 3.8-7.7 mg/l as Cl(2) for monochloramine. The inactivation kinetics of B. subtilis spores with both ozone and monochloramine varied with pH within the range of pH 6-10 investigated. The fastest ozone and monochloramine inactivation rates were observed at pH 10 and 6, respectively. Different stocks of the same strain of B. subtilis spores had different resistance to ozone and monochloramine mainly because of discrepancies in the extent of the lag phase. B. subtilis spores might not be conservative surrogates for C. parvum oocysts for ozone disinfection at relatively low temperature mainly due to the spores having a lower activation energy compared to that for the oocysts. In contrast, the activation energy for monochloramine was comparable for both microorganisms but differences in the extent of the lag phase might result in the spores being overly conservative surrogates for the oocysts at relatively low temperature.     

Inactivation of Cryptosporidium parvum oocysts with ozone and monochloramine at low temperature

The rate of Cryptosporidium parvum inactivation decreased with decreasing temperature (1-20 degrees C) for ozone and for monochloramine applied alone as well as after pre-treatment with ozone. Synergy was observed at all temperatures studied for the ozone/monochloramine sequential disinfection scheme. The synergistic effect was found to increase with decreasing temperature. The inactivation rate with monochloramine after ozone pre-treatment was 5 times faster at 20 degrees C and 22 times faster at 1 degree C than the corresponding post-lag phase rates of inactivation with monochloramine at these temperatures when no ozone pre-treatment was applied. The CT required for achieving 2-logs of inactivation ranged from 11,400 mg min l-1 at 20 degrees C to 64,600 mg min l-1 at 1 degree C when monochloramine was applied alone. In contrast, the CT required for an overall sequential inactivation of 2-logs ranged from 721 mg min l-1 at 20 degrees C to 1350 mg min l-1 at 1 degree C when applying monochloramine after ozone pre-treatment. The presence of excess ammonia in the monochloramine solutions was not responsible for the synergy observed in ozone/monochloramine sequential disinfection.     

Fate of the pathogen indicators phage ΦX174 and Ascaris suum eggs during the production of struvite fertilizer from source-separated urine

Human urine has the potential to be a sustainable, locally and continuously available source of nutrients for agriculture. Phosphate can be efficiently recovered from human urine in the form of the mineral struvite (MgNH₄PO₄·6H₂O). However, struvite formation may be coupled with the precipitation of other constituents present in urine including pathogens, pharmaceuticals, and heavy metals. To determine if struvite fertilizer presents a microbiological health risk to producers and end users, we characterized the fate of a human virus surrogate (phage ΦX174) and the eggs of the helminth Ascaris suum during a low-cost struvite recovery process. While the concentration of phages was similar in both the struvite and the urine, Ascaris eggs accumulated within the solid during the precipitation and filtration process. Subsequent air-drying of the struvite filter cake partially inactivated both microorganisms; however, viable Ascaris eggs and infective phages were still detected after several days of drying. The infectivity of both viruses and eggs was affected by the specific struvite drying conditions: higher inactivation generally occurred with increased air temperature and decreased relative humidity. On a log-log scale, phage inactivation increased linearly with decreasing moisture content of the struvite, while Ascaris inactivation occurred only after achieving a minimum moisture threshold. Sunlight exposure did not directly affect the infectivity of phages or Ascaris eggs in struvite cakes, though the resultant rise in temperature accelerated the drying of the struvite cake, which contributed to inactivation.     

Mesophilic and thermophilic anaerobic digestion of primary and secondary sludge. Effect of pre-treatment at elevated temperature

Anaerobic digestion is an appropriate technique for the treatment of sludge before final disposal and it is employed worldwide as the oldest and most important process for sludge stabilization. In general, mesophilic anaerobic digestion of sewage sludge is more widely used compared to thermophilic digestion. Furthermore, thermal pre-treatment is suitable for the improvement of stabilization, enhancement of dewatering of the sludge, reduction of the numbers of pathogens and could be realized at relatively low cost especially at low temperatures. The present study investigates (a) the differences between mesophilic and thermophilic anaerobic digestion of sludge and (b) the effect of the pre-treatment at 70 degrees C on mesophilic and thermophilic anaerobic digestion of primary and secondary sludge. The pre-treatment step showed very positive effect on the methane potential and production rate upon subsequent thermophilic digestion of primary sludge. The methane production rate was mostly influenced by the pre-treatment of secondary sludge followed by mesophilic and thermophilic digestion whereas the methane potential only was positively influenced when mesophilic digestion followed. Our results suggest that the selection of the pre-treatment duration as well as the temperature of the subsequent anaerobic step for sludge stabilization should depend on the ratio of primary to secondary sludge.