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The impact of ferrous ion reduction of chlorite ion on drinking water process performance.

The use of chlorine dioxide (ClO2) as a primary disinfectant and pre-oxidant in drinking water treatment is being explored as an alternative to chlorine for reducing disinfection by-product formation and to assure compliance with United States Environmental Protection Agency's Stage 1 Disinfection/ Disinfection By-Products Rule. However, the ClO2 by-product chlorite ion (ClO2-) is also regulated by the same regulation. Ferrous iron (Fe(II)) has been shown to effectively reduce chlorite ion to chloride ion (Cl-) and this study was conducted to evaluate the impact on overall treatment process performance due to the ferric hydroxide solids that form from the reaction. Ferrous iron application was explored at three different points in a pilot-scale water treatment system: pre-rapid mix. pre-settling and pre-filter. Chlorite ion concentrations were effectively reduced from 2mg/L to less than 0.3mg/L using an Fe(II) dose of approximately 6mg/L for all trials. Fe(II) addition at the rapid mix caused no adverse effects and, in fact, allowed for reduction of the alum dose due to the newly formed ferric hydroxide acting as a supplemental coagulant. An increase of 241 and 247% of total suspended solids influent to the filter process was observed when Fe(II) was applied at the pre-settling and pre-filter locations. Pilot-scale filter runs during these trials were less than 2 h and never obtained true steady state conditions. Jar testing was performed to better understand the nature of the ferric hydroxide solids that are formed when Fe(II) was oxidized to Fe(III) and to explore the effectiveness of Fe(II) addition at intermediate stages in the flocculation process.

Chlorides↗

Evaluation of conditioning responses of thermophilic-mesophilic anaerobically and mesophilic aerobically digested biosolids using rheological properties.

One of the most crucial processes in biosolids management is stabilisation, which is typically accomplished using either anaerobic or aerobic digestion processes. Although there are many advantages to both of these methods--such as reduction of volatile organic matter, pathogen content, and offensive odours--one common disadvantage is that digested biosolids exhibit poor dewaterability characteristics. The deterioration in dewaterability of digested biosolids leads to increases in polymer requirements, connoting higher conditioning costs. Many studies have examined this phenomenon based on conventional filterability tests such as capillary suction time and specific resistance to filtration. However, these test methods are limited in their ability to predict full-scale dewatering behaviour (particularly in centrifugation), and do not regard handling properties as being important for sludge transport. For this purpose, new rheometric methods may assist in predicting sludge behaviour, potentially allowing optimisation of stabilisation and conditioning processes relative to the desired sludge properties. This paper investigates the possible utility of these methods. Classical methods for characterising biosolids, such as filtration tests, are compared with yield stress values and other rheological properties as determined using the steady-shear rate sweep test. The rheological behaviours are compared for different sludge types, including raw and polymer conditioned thermophilic anaerobically, mesophilic anaerobically, and mesophilic aerobically digested biosolids. The paper demonstrates the determination of specific energy values associated with breakdown of the flocculated matrix. Robust modelling approaches are employed to optimise the conditioning of the digested samples, and to shed light on commonly used rheological models such as Bingham, Herschel-Bulkley, and Ostwald equations.

Aerobiosis↗

Measurement of ATP synthesis and flocculent matrix densities in mitochondria as a function of 'in vitro' ischemia in the heart and liver of rats.

We intended to determine the levels of adenosine triphosphate (ATP) synthesis at the time when mitochondria ultrastructurally show flocculent densities in the matrix space. For this purpose, mitochondria were isolated from rat heart and rat liver after the tissues have been maintained under controlled ischemic conditions in vitro at 37 degrees C for intervals of 15, 30, 45, 60, 120, 180, and 240 (heart) min. The isolated mitochondria were tested for new ATP synthesis by luciferin/luciferase luminescence in the presence of substrate and adenosine 5'-diphosphate (ADP). The luminescence peaks were standardized and related to an external measure by measuring absorbance of ATP at 259 nm where the extinction coefficient is 15,400. Mitochondrial yield was monitored by measuring succinate dehydrogenase activity in the first homogenate and in the final mitochondrial pellet. Alternatively, cytochrome oxidase activity was used and the protein in the mitochondrial pellet was also determined. We found that the yield of mitochondria was above 53-54% in both liver and heart at 2 h of ischemia. Longer intervals were accompanied by lower yields. The ability to synthesize new ATP declined at different time intervals in ischemia of the heart compared to the liver. After 30 min ischemia, the synthesis in heart mitochondria is 18% of control, while the synthesis of liver mitochondria reaches 16% of control after 45 min of in vitro ischemia. Flocculent densities in heart mitochondria appeared at 45 min ischemia in vitro and in vivo, and at 60 min in liver mitochondria. We conclude that the decline of ATP synthesis is a significant early change in mitochondria and antedates the appearance of flocculent densities.

Adenosine Triphosphate↗

In vivo genotoxicity studies with 3-monochloropropan-1,2-diol.

3-Monochloropropan-1,2-diol (3-MCPD) is a contaminant of polyamine flocculants used in the production of drinking water, but more significantly for human exposure it can arise also in certain foodstuffs containing acid-hydrolysed vegetable protein. It is carcinogenic in the rat, producing tumours in males in the testes, mammary gland and the preputial gland and also kidney tumours in both sexes. It has given positive results in in vitro mutagenicity studies, but there have been no satisfactorily conducted in vivo studies in somatic cells published in the peer reviewed literature. As a result, and because of the absence of appropriate in vivo evidence, several international regulatory agencies had previously judged it prudent to assume that 3-MCPD possessed mutagenic activity in vivo and considered 3-MCPD to be a genotoxic carcinogen. We present in this paper results from two in vivo mutagenicity studies with 3-MCPD, namely a bone marrow micronucleus test in the rat and unscheduled DNA synthesis in the rat liver, both conducted in accordance with relevant OECD protocols. These studies show that 3-MCPD does not possess genotoxic activity in vivo in the tissues examined. On the basis of these findings, and along with evidence that tumours may be induced by mechanisms involving either hormonal disturbances or sustained cytotoxicity, we believe that 3-MCPD may now be considered to be carcinogenic to rodents via a non-genotoxic mechanism.

Animals↗

Diphtheria toxin resistance in human lymphocytes and lymphoblasts in the in vivo somatic cell mutation test.

It has been shown that circulating peripheral blood lymphocytes can be used for the enumeration of 6-thioguanine-resistant cells that presumably arise by mutation in vivo. This somatic cell mutation test has been studied in lymphocytes from human populations exposed to known mutagens and/or carcinogens. The sensitivity of the test could be further enhanced by including other gene markers, since there is evidence for locus-specific differences in response to mutagens. Resistance to diphtheria toxin (Dipr) seemed like a potential marker to incorporate into the test because the mutation acts codominantly, can readily be selected in human diploid fibroblasts and Chinese hamster cells with no evidence for cell density or cross-feeding effects, and can be assayed for in nondividing cells by measuring protein synthesis inhibition. Blood samples were collected from seven individuals, and fresh, cryopreserved, or Epstein-Barr virus (EBV)-transformed lymphocytes were tested for continued DNA synthesis (3H-thymidine, autoradiography) or protein synthesis (35S-methionine, scintillation counting). Both fresh and cryopreserved lymphocytes, stimulated to divide with phytohemagglutinin (PHA), continued to synthesize DNA in the presence of high doses of diphtheria toxin (DT). Similarly, both dividing (PHA-stimulated) and nondividing fresh lymphocytes carried on significant levels of protein synthesis even 68 hr after exposure to 100 flocculating units (LF)/ml DT. The findings were confirmed in cord blood lymphocytes, ruling out the possibility that diphtheria immunization could have led to a selection of Dipr lymphocytes. One lymphoblast line (EBV-transformed lymphocytes) showed a reduction in protein synthesis to 0.2% of controls only at 192 hr after exposure to 100 LF/ml. The results suggest that human T and B lymphocytes may not be as sensitive to DT protein synthesis inhibition as human fibroblast and Chinese hamster cells. For this reason, Dipr may not be a suitable marker for the somatic cell mutation test.

Cells, Cultured↗

A new in-vitro agglutination technique for potency estimation of antisnake venom serum (ASVS).

Traditionally the potency of ASVS is assayed quantitatively by in-vivo neutralization test for lethality in mice. A sensitive and simple in-vitro agglutination assay for the quantitative determination of Antisnake Venom Serum (ASVS) potency is reported. The method is rapid, cheap, simple, economical and above all does not require the use of experimental animals for potency assay of in process, unpurified and purified sera batches. Among in-vitro procedures, agglutination assay was favored in comparison to flocculation as the later was found to give variable results and also time consuming (high Kf value). Before application, the method was standardized and validated for choice and concentration of particulate material (latex vs. bentonite), temperature and optimum antiserum concentration. It is well known fact that venoms lose toxicity on dilution however this study demonstrated that the bentonite adsorbed venoms of the entire four snake species viz., Cobra, Krait, Russell's viper and Echis are stable even up to 30 days of storage. Among five lots each of unpurified serum, unprocessed plasma and purified sera tested, the results were found comparable with universally accepted in-vivo biological assay. The coefficient of correlation was found to be near 1.0 within 95% fiducial limits of acceptance and also significantly less variation was observed in the mean potency values and standard deviations. For all results p value was observed to be <0.01. Results indicate that in-vitro agglutination assay is suitable and can be used for potency estimation of in process as well as unpurified and purified ASVS batches.

Adsorption↗

[The toxicity variation of organic extracts in drinking water treatment processes].

Source water samples and outlet water samples from different treatment processes of the Beijing Ninth Water Works were concentrated in situ with XAD-2 filled columns. GC-MS analysis and toxic assessment including acute toxicity evaluation by luminescent bacterium bioassay(Q67 strains) and mutagenicity assessment by Ames test(TA98 and TA100 strains with and without S9 addition) were conducted on these samples. The results showed that prechlorination caused the direct and indirect frame shift mutagenicity as well as indirect base pair substitute mutagenicity. Addition of coagulant may increase the base pair substitute mutagenic effects greatly. Sand and coal filtration and granular activated carbon filtration could effectively remove most of the formed mutagens. The rechlorination do not obviously increase the mutagenic effects. No mutagenic effect was observed in tap water. Acute toxicity showed the same variation with that of mutagenicity during the treatment processes. Sample from flocculation treatment process was found to be the most toxic sample. Results of GC-MS analysis showed that water in this plant was not contaminated by PCB. Concentrations of toluene, naphthalene and phenol increased in flocculation treatment process and in tap water. However, the concentrations of these substances were at the level of microgram/L, therefore, were not high enough to cause mutagenicity.

Gas Chromatography-Mass Spectrometry↗

STA10 repression of STA gene expression is caused by a defective activator, flo8, in Saccharomyces cerevisiae.

The expression of STA genes that encode extracellular glucoamylase isozymes is repressed in most laboratory Saccharomyces cerevisiae strains, which are believed to contain an undefined repressor, designated STA10. To identify the regulator involved in STA10 repression, we investigate the FLO8, MSN1, MSS11, STE12, and TEC1 genes. The Deltaflo8 or Deltamss11 deletion mutants in the sta10 genetic background exhibit both a loss of flocculation ability and a reduction in extracellular glucoamylase activity, as in the STA10 strain. Moreover, the STA10 repression is suppressed completely or partially by the introduction of a single copy of the FLO8 or MSS11 genes. Sequence analysis and complementation testing of the STA10 strain reveal that it has an inactive, mutated flo8-1 allele. A random spore analysis and transplacement (allele replacement) experiment confirms that the repressive phenotype of STA10 is due to the amber mutation of the transcriptional activator, FLO8.

Alleles↗

Hydrodynamic drag on non-spherical floc and free-settling test.

This work numerically investigates the hydrodynamic drag force exerted on a porous spheroid floc moving steadily through a quiescent Newtonian fluid over a wide range of the Reynolds number. The flow patterns for a highly porous spheroid moving at an elevated Reynolds number are basically the same as those at a low Reynolds number, which extends the applicable range of a creeping-flow based correlation to the higher Reynolds number regime. The shape effect becomes more prominent as the spheroid becomes more porous. Using the equivalent diameter, defined as the geometric mean diameter of the principal axes, leads to a universal correlation relating to the drag force, aspect ratio, and interior permeability. In addition, free-settling experiments are performed to estimate how the non-spherical shape affects the three sludge samples. The possible errors in data reduction for the free-settling test are attributed to the a/b ratio and the internal permeability. The errors range from 16-34% for a/b = 0.6-2.0.

Flocculation↗

Separation characteristics of liquid nematode cultures and the design of recovery operations.

Production of nematode-based pesticides involves the recovery of a viable nematode life stage known as the infective juvenile (IJ) from fermentation broth. Waste components to be separated from the IJs include non-IJ life stages, dead nematodes, nematode debris, spent media, and the nematode's associated bacteria. This paper reports separation characteristics of liquid cultures and suspensions of the nematodes Phasmarhabditis hermaphrodita, Steinernema feltiae, and Heterorhabditis megidis measured at small scale. Separation characteristics were determined for dead-end filtration, gravity settling and flotation. Results were used to identify large-scale recovery procedures. Separation of culture liquid by dead-end filtration of the crude fermentation broth was not possible due to rapid blinding of filters. However, nematode-water suspensions prepared by gravity settling could be concentrated using this separation method. Settling tests indicated that IJs could be efficiently separated from culture liquid by centrifugation but not by gravity settling. Examination of the effects of nematode concentration indicated an optimum concentration for gravity settling that may entail modest dilution of the fermentation broth. Flocculation of insoluble spent media in suspensions of P. hermaphrodita prevented its separation from nematodes by gravity settling. However, attachment of air bubbles to spent media allowed removal by flotation. Finally, adjustment of continuous phase density using sucrose allowed separation of non-IJ life stages, dead nematodes, and discarded cuticles from the IJs by flotation. The efficiency of this separation decreased with increasing nematode-solute contact time.

Animals↗

Monitoring of coagulation performance and determination of coagulant dosage using a pilot in-line filter.

A rapid method using the pilot in-line filter to detect any change in coagulation performance was a proposed in this study. This method attempted to detect a change in coagulant dosage and mixing intensity by evaluating the filtrate quality of the in-line filter, which took the rapidly mixed water. Since the response time of this method was less than 10 min, it could be valuable to monitor the coagulation performance. The in-line filter was found more useful without underdrain. The in-line filter was more sensitive to a change in filtrate quality without underdrain than with underdrain. A new method, which combines a jar test with the in-line filter, was proposed to determine the coagulant dosage. This method reflected the actual plant situation more accurately than a jar test.

Aluminum Hydroxide↗

Giardia and Cryptosporidium in source waters of São Paulo State, Brazil.

Giardia lamblia and Cryptosporidium parvum are two protozoan intestinal parasites responsible for many drinking-water-related disease outbreaks in recent years. They are very resistant to conventional water treatment processes, can persist for long times in the environment and are, therefore, of great concern for public health. This work aimed to evaluate the presence of Giardia and Cryptosporidium in water sources from São Paulo State, Brazil, as part of the "Evaluation of Inland Waters from São Paulo State" project from CETESB. Over a period of 19 months, 278 water samples from 28 sites located in 10 watersheds were analysed. The immunofluorescence assay was used after concentration of the samples by the calcium carbonate flocculation technique. Thermotolerant (faecal) coliforms, faecal streptococci and Clostridium perfringens were also determined in order to verify the existence of correlation between these bacterial indicators and the protozoa. Giardia and Cryptosporidium were detected in 27% and 2.5% of the samples, respectively, a lower figure compared with the results reported by other authors, especially for Cryptosporidium. A Spearman rank correlation test demonstrated a significant correlation between Giardia and faecal indicator concentrations. According to the American Regulation of Monitoring (ICR), treated water from 16 of these 28 collection sites should also be analysed to evaluate whether the treatment process could remove the parasites. Some technical deficiencies of these methods still limit the utilisation of the monitoring results for public health decisions, but the data here reported will help to improve the quality of drinking water in São Paulo State.

Animals↗

Synthesis and characterization of active ester-functionalized polypyrrole-silica nanoparticles: application to the covalent attachment of proteins.

Novel ester-functionalized polypyrrole-silica nanocomposite particles were prepared by oxidative copolymerization of pyrrole and N-succinimidyl ester pyrrole (50/50% initial concentrations), using FeCl3 in the presence of ultrafine silica nanoparticles (20 nm diameter). The N-succinimidyl ester pyrrole monomer was prepared in aqueous solution using 1-(2-carboxyethylpyrrole) and N-hydroxysuccinimide in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. The resulting nanocomposites (N-succinimidyl ester polypyrrole-silica) are raspberry-shaped agglomerates of silica sol particles "glued" together by the insoluble poly(pyrrole-co-N-succinimidyl pyrrole). The N-succinimidyl ester polypyrrole-silica particles were characterized in terms of their size, density, copolymer content, and polydispersity. Scanning electron microscopy and disk centrifuge sedimentometry confirmed that the nanocomposite particles had narrow size distributions. X-ray photoelectron spectroscopy analysis indicated a silica-rich surface and a high surface concentration of N-succinimidyl ester groups. These nanoparticles exhibited good long-term dispersion stability. The chemical stability of the ester functions in aqueous media after several weeks of storage was monitored by FTIR spectroscopy. The functionalized nanocomposites were tested as bioadsorbents of human serum albumin (HSA). The very high amount of immobilized HSA determined by UV-visible spectroscopy is believed to be due to covalent binding. Incubation of the HSA-grafted nanocomposite with anti-HSA resulted in immediate flocculation, an indication that they are alternative candidates for visual diagnostic assays.

Esters↗

Behaviour of heavy metals during tertiary bio-filtration.

More stringent water quality parameters in the Annex of the Water Framework Directive 2000/60/EC led to the introduction of "Maximum Tolerable Risk concentrations" (MTR-values) in the national legislation in The Netherlands (Vierde Nota Waterhuishouding). The MTR-values give limitations for the concentrations of e.g. heavy metals (HM's) but also for nutrients: Ntot < 2.2 mg/l, Ptot <0.15 mg/l. Investigations of HM removal during denitrifying flocculation filtration are conducted on the effluent of a typical modern Dutch WWTP. Because of low concentrations of HM's in the feed water, a cocktail of copper, nickel and zinc chloride (approximately equal 150-200 microg/l) is dosed before filtration. Preliminary jar tests and filtration tests with media filtration in pilot-scale and lab-scale are conducted. The results show high removal of nickel and zinc during jar tests with dosing of powdered activated carbon. During filtration tests at pilot-scale the bounded fraction of copper and zinc is highly removed. All three HM's are removed in the lab-scale activated carbon filter. After dosing, nickel is found mainly in the dissolved form, but it is removed in the lab-scale activated carbon filter. The removal of HM's via adsorption subsequently leads to a discussion on the toxicity of HM's and their bio-availability.

Filtration↗

Municipal wastewater sludge dewaterability and the presence of microbial extracellular polymer.

Dewatering of sewage sludge is an essential and costly part of the wastewater treatment process. The presence of microbial extracellular polymer (ECP) is important for sludge flocculation, but ECP has also been shown to have a detrimental effect on the dewaterability of certain sludge types. This paper investigates the relationship between sludge dewaterability and the level of ECP present in a range of sludges obtained from 8 full-scale municipal treatment works in the UK. Sludge dewaterability was determined using the capillary suction time (CST) test, and a thermal extraction process followed by solvent precipitation was used for ECP extraction. The results indicate that for each type of sludge examined there appears to be an optimum level of ECP (raw sludge 20 mg ECP/g SS; activated sludge 35 mg ECP/g SS; digested sludge 10 mg ECP/g SS) at which the sludge should exhibit maximum dewaterability. The establishment of a trend between sludge dewaterability and the quantity of ECP present opens up the possibility of manipulating the level of microbial polymer present to aid sludge dewatering, and hence reduce plant operating costs.

Flocculation↗

Improved method and test strategy for recovery of enteric viruses from shellfish.

An improved recovery method and testing strategy were devised for recovery of low numbers of enteric viruses from each of three commercially important shellfish species. Effective recovery of virus depended as much upon details of the test strategy adopted for use of the improved method with each species as on the method itself. The most important test details involved sample composition, pool size, and method of use of cell cultures. Recovery sensitivity measured permitted detection of 25 to 3 plaque-forming units of enteroviruses and 100 to 27 plaque-forming units of reovirus through their recovery in cell culture, with effectivenesses averaging 64 and 46%, respectively. Test samples prepared by the improved recovery method were virtually cytotoxicity free. Optimal recovery of virus on 45-cm2 cell culture monolayers was obtained with 1-ml inocula adsorbed for 2 h. The most effective recovery of virus from shellfish samples was made by a sequential adsorption procedure which allowed equal exposure of an entire sample to each of two or more cell cultures. Removal of nonviral contaminants from test samples by antibiotic treatment was preferable to the use of ether or membrane filtration procedures.

Animals↗

Intermittent vs continuous operation of upflow anaerobic sludge bed reactors for dairy wastewater and related microbial changes.

This work compares continuous vs intermittent UASB reactors inoculated with flocculent sludge for the treatment of dairy effluents. The effects of effluent recirculation on the performance of intermittent reactors were assessed as well as the differences in specific methanogenic activity (SMA) with different substrates for the biomass from continuous and intermittent UASB reactors. Compared to the continuous operation the intermittent operation resulted in higher methanization of the removed COD (64-78% and 65-88%, respectively) whilst the effluent recirculation presented beneficial effects when applied during the stabilization period and was clearly detrimental when applied during the feed period of the intermittent operation. The SMA tests showed that the intermittent operation causes a shift in the microbial populations towards a better adaptation for the degradation of complex substrates confirmed by the meaningfull contribution of methane production through a pathway other than acetoclastic methanogenesis observed in the biomass taken from intermittent UASB reactors.

Anaerobiosis↗

Proof-of-concept of a novel micro-bioreactor for fast development of industrial bioprocesses.

The experimental performance of a novel micro-bioreactor envisaged for parallel screening and development of industrial bioprocesses has been tested in this work. The micro-bioreactor with an internal volume of 4.5 mL is operated under oscillatory flow mixing (OFM), where a controllable mixing and mass transfer rates are achieved under batch or continuous laminar flow conditions. Several batch fermentations with a flocculent Saccharomyces cerevisiae strain were carried out at initial glucose concentrations (S(0)) range of approximately 5-20 g/L and compared to yeast growth kinetics in a stirred tank (ST) bioreactor. Aerobic fermentations were monitored ex situ in terms of pH, DO, glucose consumption, and biomass and ethanol production (wherever applicable). An average biomass production increase of 83% was obtained in the micro-bioreactor when compared with the ST, with less 93.6% air requirements. It also corresponded to a 214% increase on biomass production when compared with growth in a shaken flask (SF) at S(0) = 20 g/L. Further anaerobic fermentations at the same initial glucose concentration ranges gave the opportunity to use state-of-the-art fiber optics technology for on-line and real-time monitoring of this bioprocess. Time profiles of biomass concentration (measured as optical density (OD)) were very similar in the ST bioreactor and in the micro-bioreactor, with a highly reproducible yeast growth in these two scale-down platforms.

Biomass↗