Elimination of human enteric viruses during conventional waste water treatment by activated sludge

The present study was undertaken to determine if viruses were selectively eliminated during waste water treatment. Human enteric viruses were detected at all steps of treatment in a conventional activated sludge waste water treatment plant. Liquid overlays and large volume sampling with multiple passages on BGM cells permitted the detection of poliovirus (serotypes 1, 2, and 3), coxsackievirus B (serotypes 1, 2, 3, 4, and 5), and echovirus (serotypes 3, 14, and 22), as well as reoviruses. The mean virus concentration was 95.1 most probable number of infectious units per litre (mpniu/L) in raw sewage, 23.3 in settled water, 1.4 in effluent after activated sludge treatment, and 40.3 mpniu/L in sludge samples. All samples of raw sewage and settled water, 79% of effluent water, and 94% of sludge samples contained viruses. The mean reduction was 75% after settling and 98% after activated sludge treatment. Poliovirus type 3 was rarely isolated after the activated sludge treatment, but was still detected in about one-third of the sludge samples. Reoviruses and coxsackieviruses were detected at similar rates from all samples and appear to be more resistant to the activated sludge treatment than poliovirus type 3. Poliovirus types 1 and 2 were present in almost every sample of raw sewage and settled water and still found in about half of the effluent and sludge samples, indicating a level of resistance similar to that of reoviruses and coxsackieviruses.     

Assessment of recovery efficiency of beef extract reagents for concentrating viruses from municipal wastewater sludge solids by the organic flocculation procedure

This study was designed to assess the capacity of beef extract reagents to form flocs suitable for virus adsorption. Reagent comparisons resulted in the establishment of a modified organic flocculation procedure to concentrate viruses desorbed from sewage sludge solids with currently available modified powdered beef extracts. The method, based on supplementation with paste beef extract floc, achieved virus recoveries comparable to those obtained with powdered beef extract produced before a 1979 change in the manufacturing process. When primary settled sludge solids originating from mostly domestic waste were eluted with an unsupplemented modified powdered beef extract, high virus recovery efficiency was observed upon concentration by organic flocculation. This appreciable increase might have been due to floc-forming substances that were present in the primary settled sludge. These substances did not appear to be present in settled sludge collected from biologically treated wastes. Apparently, the floc-forming substances had been either removed or substantially altered during biological treatment.     

Assessment of recovery efficiency of beef extract reagents for concentrating viruses from municipal wastewater sludge solids by the organic flocculation procedure.

This study was designed to assess the capacity of beef extract reagents to form flocs suitable for virus adsorption. Reagent comparisons resulted in the establishment of a modified organic flocculation procedure to concentrate viruses desorbed from sewage sludge solids with currently available modified powdered beef extracts. The method, based on supplementation with paste beef extract floc, achieved virus recoveries comparable to those obtained with powdered beef extract produced before a 1979 change in the manufacturing process. When primary settled sludge solids originating from mostly domestic waste were eluted with an unsupplemented modified powdered beef extract, high virus recovery efficiency was observed upon concentration by organic flocculation. This appreciable increase might have been due to floc-forming substances that were present in the primary settled sludge. These substances did not appear to be present in settled sludge collected from biologically treated wastes. Apparently, the floc-forming substances had been either removed or substantially altered during biological treatment.     

Malachite green-INT (MINT) method for determining active bacteria in sewage.

A membrane filtration method was developed to determine the proportion of active (respiring) bacteria at various stages of sewage treatment. Samples were incubated in the presence of 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) and, after fixation, passed through membrane filters. Filters were counterstained with malachite green and then were examined by bright-field microscopy. The contrast between bacteria and the filter background was greatly improved by drying and then clearing the filter before counterstaining. By this method, it was found that active bacterial fractions in raw sewage, settled sewage, and secondary effluent were 40, 29, and 58%, respectively, whereas the proportion of respiring bacteria in chlorinated secondary effluent was 6.1%. The active bacterial fraction of activated sludge was found to be 16%. The proposed method represents a significant improvement in speed and simplicity over existing methods for determining active bacteria in sewage.     

Some effects of organic pollution on the distribution and seasonal incidence of Chironomidae in riffles in the River Cole

SUMMARY. The effects of a sewage effluent discharge into the River Cole, Birmingham, were determined by regular collections of water and biological samples from six comparable stations, one above and five below the sewage effluent. The effluent reduced the dissolved oxygen concentration and increased the ammonia, orthophosphate and BOD concentrations in the water. The effects were much more severe during the summer.
The Chironomidae were very important constituents of the benthic macroinvertebrate community at the stations downstream of the effluent. Larval densities of over 30,000 m⁻² were recorded. Eleven species were common and of these Chironomus riparius was the most tolerant. There was considerable variation in the seasonal incidence of the different species. Brillia longifurca was most abundant in the spring while Cricotopus bicinctus was most abundant in the summer. Brillia longifurca, Paratrichocladius rufiventris, Prodiamesa olivacea, Micropsectra atrofasciata and Eukiefferiella claripennis had three peaks in numbers of larvae during the year.
The increase in the polluted state of the river in the summer resulted in changes in the chironomid populations. Some were suppressed, e.g. E. claripennis , while C. riparius extended its range and achieved maximum abundance further downstream during the summer.     

Activity of microorganisms in activated sludge in relation to the concentration of mercury in sewage

Activated sludge from the refinery of a plant producing synthetic rubber was shown to adsorb mercury ions contained in the sewage. As a result, the content of microorganisms in the sludge as well as the activity of dehydrogenase decreased. The quantity of Pseudomonas aeruginosa cells was 40 x 10(6) per 1 ml in a chemically defined medium without mercury ions, 20 x 10(5) per 1 ml in the water after the first sedimentation tank (0.07 to 0.08 mg Hg per litre), and 10 x 10(6) per 1 ml in the water of sewage common to the whole plant (0.13 to 0.14 mg Hg per litre). The use of the phage-resistant P-aeruginosa T-76 culture adapted to synthetic organic pollutants contained in the sewage of the plant together with the activated sludge of the refinery increased the biological activity of the sludge and improved the quality of purification in terms of dehydrogenase activity and chemical uptake of oxygen.     

Biological Uptake of Phosphorus by Activated Sludge

The ability of activated sludge to remove phosphates was studied by adding carrier-free (32)P to raw sewage and measuring incorporation of the radioactivity into the cells over a period of time. Radioisotope determinations indicated that 48% of the (32)P radioactivity was removed by 12 hr. However, chemical methods indicated that only 30% of the orthophosphate apparently disappeared from the sewage during this period. Experiments with sludge prelabeled with (32)P indicated that considerable phosphate turnover occurred. The cells released large amounts of radioactivity as they were incorporating fresh phosphates. Starvation in isotonic saline for 18 hr caused the sludge to dump phosphate. When introduced into fresh sewage containing (32)P, the starved sludge removed about 60% of the radioactivity in 6 hr with little phosphate turnover. The ability of sludge to remove (32)P was inhibited approximately 83% by 10(-3)m 2,4-dinitrophenol. This inhibition was at the expense of the cell fraction that contained ribonucleic acid and deoxyribonucleic acid. The sludge cells released orthophosphate when exposed to the chemical agent. Experiments using (45)Ca indicated that calcium phosphate precipitation plays a minor role in phosphate removal under our experimental conditions.     

Spatial distribution of dynamics characteristic in the intermittent aeration static composting of sewage sludge

Spatial differences and temporal changes in biological activity characteristics were investigated in a static reactor using intermittent aeration during the sewage sludge composting process. Pumice was proposed as a bulking agent in the composting of sewage sludge. Variations in temperature, moisture, oxygen level, volatile solids, specific oxygen uptake rate (SOUR) and dehydrogenase activity (DHA) were determined during 28 days of composting. The peak temperature in the upper region of the reactor was 10 °C higher than that at the bottom. The moisture level in the middle region was significantly higher than that of other positions. Analysis of SOUR and DHA indicated that the lowest level of sludge stability was at the bottom region. These spatial and temporal differences in biochemical dynamics in the static system could extend the composting period and affect product uniformity.     

The effect of sewage effluent on Grasmere (English Lake District) with particular reference to inorganic nitrogen transformations

SUMMARY. In the summer of 1971 the village of Grasmere was converted from septic tank to mains drainage with sewage treatment at an activated sludge plant. The effluent was discharged into the River Rothay, the main inflow of a nearby small lake (Grasmere). This paper describes some of the effects on the lake. The mean areal hypolimnetic oxygen deficit increased from 274 to 434 mg O₂ m⁻² day⁻¹ with an accompanying marked increase in the degree of deoxygenation in the hypolimnion. Plate counts of bacteria in the surface water increased for 2 years but this increase was not sustained. The mean summer soluble reactive phosphorus concentration did, however, increase significantly ( P = 0.05), but the same was not true of nitrate levels. This paper is particularly concerned with inorganic nitrogen transformations and analyses of the main inflow have shown that 50–98% of the ammonia and 10–40% of the nilrate entering in this river was derived from the sewage effluent. The concentrations in the main body of the lake were usually lower, possibly due to assimilation and denitrification in the shallower reaches of the lake. Seasonal changes in the inorganic nitrogen species in the hypolimnion showed three distinct phases of activity, ammonification, nitrification and denitrification. Nitrification accounted for approximately a quarter of the oxygen uptake in the hypolimnion.     

Hydrogen peroxide as a supplemental oxygen source for activated sludge: Microbiological investigations

Summary Parallel bench-scale activated sludge systems were operated using air or hydrogen peroxide as oxygen source. The use of H₂O₂ resulted in a temporary decrease of COD reduction, an increase of the catalase activity of the activated sludge, a depression of the nitrification, and a marked decrease of some filamentous organisms. Enumeration of some microbiologic groups indicated that the counts of enterobacteria, coliforms, staphylococci, and streptococci were lower in the H₂O₂ unit than in the parallel air unit. Also the use of H₂O₂ did not induce the selection of bacterial species that are more resistant to H₂O₂. The increase in catalase activity after H₂O₂ addition might be the result of a stimulation of catalase synthesis in catalase positive microorganisms.List of Abbreviation COD chemical oxygen demand, mg O₂/1 - COD_eff chemical oxygen demand of the effluent, mg O₂/1 - DO dissolved oxygen, mg O₂/1 - MLSS mixed liquor suspended solids, g dry weight/1 - SVI sludge volume index, ml settled sludge per liter/MLSS (ml/g) - F:M sludge loading factor or the Food to Microorganisms ratio, g COD/g MLSS.day     

Seasonal variation in the properties of titania photocatalysts produced from Ti-salt flocculated bioresource sludge

Ti-salt flocculation of biologically treated sewage effluent (BTSE) was carried out on monthly basis during one year to trace the seasonal variation in the properties of BTSE, Ti-salt flocculated BTSE and titania photocatalysts. Titania photocatalysts were produced from incineration of Ti-salt flocculated sludge at 600 °C. The physio-chemical properties of BTSE, Ti-salt flocculated BTSE and titania photocatalysts were investigated. The photocatalytic activity of titania was examined using different substrates of rhodamine B and humic acid under UV light irradiation. Results indicated that the flocculation performance of Ti-salt was not affected by the seasonal variation of BTSE. BTSE characteristics resulted in marginal effect in titania characterisation and photocatalytic activity. Titania photocatalysts produced from Ti-salt flocculated sludge in different seasons showed constant anatase phase, high BET surface area and high photocatalytic activity.     

Potential utilisation of sewage sludge and paper mill waste for biosorption of metals from polluted waterways

The adsorption of cadmium, copper(II), lead and zinc ions from aqueous solution by sewage sludge, paper mill waste (PMW) and composted PMW was investigated along with the influence of pre-treatment on composted PMW. Langmuir adsorption isotherms were fitted where appropriate. Sewage sludge was the most effective biosorbent of the waste products for all metal ions examined, adsorbing, for example, up to 39.3 mg/g of Pb at an initial concentration of 77.8 mg/l. PMW was a less effective biosorbent than sewage sludge. However, it was found that composting the PMW resulted in an increase in metal uptake capacity and both sewage sludge and composted PMW have potential for low-cost remediation of high leachate wastewaters. The desorption of metal ions from PMW compost was most effective using 0.1 N H2SO4 and 1 mM nitrilotriacetic acid (NTA).     

Genotoxicity of wastewater samples from sewage and industrial effluent detected by the Allium root anaphase aberration and micronucleus assays.

The genotoxicity of wastewater samples from sewage, and industrial effluent from the Amritsar, India, area were investigated using the Allium micronucleus and anaphase aberration assays. Raw sewage samples and acetone extracts of the dehydrated sewage were use for treatment of the Allium roots. Industrial effluents were collected and stored in the form of sludge (semi-dried matter). The acetone extracts of the sludge samples were also used for treatment of the Allium roots. From the Allium root micronuclei tests on the sewage extracts, no significant increase in the number of micronuclei was found in comparison with negative controls. All the other extracts from industrial effluent showed positive responses both in the micronucleus and anaphase aberration assays.     

The fate of crop nutrients during digestion of swine manure in psychrophilic anaerobic sequencing batch reactors

The objectives of the study were to measure the levels of manure nutrients retained in psychrophilic anaerobic sequencing batch reactors (PASBRs) digesting swine manure, and to determine the distribution of nutrients in the sludge and supernatant zones of settled bioreactor effluent. Anaerobic digestion reduced the total solids (TS) concentration and the soluble chemical oxygen demand (SCOD) of manure by 71.4% and 79.9%, respectively. The nitrogen, potassium, and sodium fed with the manure to the PASBRs were recovered in the effluent. The bioreactors retained on average 25.5% of the P, 8.7% of the Ca, 41.5% of the Cu, 18.4% of the Zn, and 67.7% of the S fed to the PASBRs. The natural settling of bioreactor effluent allowed further nutrient separation. The supernatant fraction, which represented 71.4% of effluent volume, contained 61.8% of the total N, 67.1% of the NH4-N, and 73.3% of the Na. The settled sludge fraction, which represented 28.6% of the volume, contained 57.6% of the solids, 62.3% of the P, 71.6% of the Ca, 89.6% of the Mg, 76.1% of the Al, 90.0% of the Cu, 74.2% of the Zn, and 52.2% of the S. The N/P ratio was increased from 3.9 in the raw manure to 5.2 in the bioreactor effluent and 9.2 in the supernatant fraction of the settled effluent. The PASBR technology will then substantially decrease the manure management costs of swine operations producing excess phosphorus, by reducing the volume of manure to export outside the farm. The separation of nutrients will also allow land spreading strategies that increase the agronomic value of manure by matching more closely the crop nutrient requirements.     

构树的污泥适应性及养分和重金属吸收累积特征

构树是我国重要速生经济树种,具有适应性强、生物量大和重金属富集能力强等优点,而污泥中含有大量养分和重金属,在污泥中种植构树有望同时实现污泥生态修复和构树资源生产。本研究通过盆栽试验,分析在对照(赤红壤)、50%污泥(污泥、赤红壤混合基质,重量比各50%)和100%污泥基质中构树生长及不同部位(根、茎、叶)养分和重金属吸收累积特征,并通过主成分分析和隶属度函数对吸收累积能力进行综合评价。结果表明: 构树在50%和100%污泥中均可正常生长且株高、生物量显著高于对照,在100%污泥中长势最好,质量指数(1.02)分别是对照和50%污泥处理的4.3和2.4倍。50%和100%污泥处理构树各部位N含量和茎P含量显著高于对照,100%污泥处理构树茎、叶K含量显著低于对照。构树对Cu、Zn、Pb、Cd、Ni的吸收部位以根为主,根系重金属含量与污泥比例呈正相关,叶Pb、Cd含量符合《饲料卫生标准》(GB 13078—2017)。构树对Cd的吸收累积效果好于其他重金属元素。与对照相比,50%和100%污泥处理构树根部Zn、Pb、Cd滞留率显著提高(57.8%~85.8%),100%污泥处理构树根部Cu、Ni滞留率显著提高(67.5%和74.8%)。污泥处理全株养分和重金属累积量均显著大于对照,其中100%污泥处理显著大于50%污泥处理。与50%污泥处理相比,100%污泥处理构树各部位及全株养分和重金属累积量大幅提高。不同处理下构树污泥适应性和元素吸收累积的综合评价得分为100%污泥(0.848)>50%污泥(0.344)>对照(0.080)。构树对污泥具有良好的适应性,在纯污泥中能够正常生长并具有较强的吸收累积养分和重金属能力,可在修复污泥的同时进行构树资源生产。     

Microbial Ecology of Activated Sludge: II. Bacteriophages, Bdellovibrio, Coliforms, and Other Organisms

A comparative estimation of the coliform population of raw sewage, activated sludge, and the effluent derived therefrom revealed that raw sewage had a preponderance of Escherichia coli (75%), as compared with 25 and 30%, respectively, in sludge and effluent. Nitrogen-free mannitol-sucrose enrichments of activated sludge resulted in the isolation of Azotobacter agilis, Aerobacter aerogenes, Corynebacterium laevaniformans, and an Achromabacter species. Sludge had a large population of C. laevaniformans and A. aerogenes but not of Azotobacter. The bacterial parasites, Bdellovibrio and bacteriophages, were not active during activated-sludge treatment. A 10-fold reduction in phage content occurred after 2 hr of aeration, but the Bdellovibrio population was unaffected.     

Metabolic Factors Affecting Enhanced Phosphorus Uptake by Activated Sludge

Activated sludges obtained from the Rilling Road plant located at San Antonio, Tex., and from the Hyperion treatment plant located at Los Angeles, Calif., have the ability to remove all of the orthophosphate normally present in Tucson sewage within 3 hr after being added to the waste water. Phosphorus removal was independent of externally supplied sources of energy and ions, since orthophosphate and (32)P radioactivity were readily removed from tap water, glass-distilled water, and deionized water. Phosphorus uptake by Rilling sludge in the laboratory appears to be wholly biological, as it has an optimum pH range (7.7 to 9.7) and an optimum temperature range (24 to 37 C). It was inhibited by HgCl(2), iodoacetic acid, p-chloromercuribenzoic acid, NaN(3), and 2, 4-dinitrophenol (compounds that affect bacterial membrane permeability, sulfhydryl enzymes, and adenosine triphosphate synthesis). Uptake was inhibited by 1% NaCl but was not affected by 10(-3)m ethylenediaminetetraacetic acid disodium salt (a chelating agent for many metallic ions).     

Tracking human sewage microbiome in a municipal wastewater treatment plant

Human sewage pollution is a major threat to public health because sewage always comes with pathogens. Human sewage is usually received and treated by wastewater treatment plants (WWTPs) to control pathogenic risks and ameliorate environmental health. However, untreated sewage that flows into water environments may cause serious waterborne diseases, as reported in India and Bangladesh. To examine the fate of the human sewage microbiome in a local municipal WWTP of Hong Kong, we used massively parallel sequencing of 16S rRNA gene to systematically profile microbial communities in samples from three sections (i.e., influent, activated sludge, and effluent) obtained monthly throughout 1 year. The results indicated that: (1) influent sewage bacterial profile reflected the human microbiome; (2) human gut bacterial community was the dominant force shaping influent sewage bacterial profile; (3) most human sewage bacteria could be effectively removed by the WWTP; (4) a total of 75 genera were profiled as potentially pathogenic bacteria, most of which were still present in the effluent although at a very low level; (5) a grouped pattern of bacterial community was observed among the same section samples but a dispersed pattern was found among the different section samples; and (6) activated sludge was less affected by the influent sewage bacteria, but it showed a significant impact on the effluent bacteria. All of these findings provide novel insights toward a mechanistic understanding of the fate of human sewage microbiome in the WWTP.     

Viruses in sewage: effect of phosphate removal with calcium hydroxide (lime).

During calcium hydroxide (lime) treatment (pH 9.6 to 10.5) of wastewaters for phosphate removal there was also a two-log removal of added poliovirus (type I, Sabin) from effluents. A similar virus reduction was seen in the sludge generated in these experiments. However, in view of the limitations of techniques for virus recovery from sludge, only a small portion of the infectious virus present in lime sludge may have been detected. Storage of lime sludge at 28 degrees C for up to 48 h produced no appreciable reduction in the virus titre. Five sets of field samples of sewage, effluents, and sludge from a sewage treatment plant (Kemptville, Ont.) which utilizes lime for phosphate removal were also examined for indigenous viruses being BS-C-1 cells. All of the sample of lime sludge and 80% of the samples of both sewage and lime-treated effluent revealed virus; after chlorination only 20% of the lime-treated effluent samples were positive for virus. In contrast, in an earlier study with essentially the same experimental set up, 76% of the sample of chlorinated primary effluent were found to contain virus. Because of the easily detectable quantities of infectious virus in lime sludge and due to the lack of virus inactivation during storage of such sludge, caution must be exercised in its handling and disposal.     

Feasibility study of a pilot-scale sewage treatment system combining an up-flow anaerobic sludge blanket (UASB) and an aerated fixed bed (AFB) reactor at ambient temperature

A feasibility test of a 17 m3-pilot-scale sewage treatment system was carried out by continuous feeding of raw municipal sewage under ambient temperature conditions. The system consisted of a UASB and an aerated fixed bed reactor. Some of the effluent from the fixed bed reactor was returned to the UASB influent in order to provide a sulfate source. The total BOD of 148-162 mg l(-1) in the influent was reduced to a more desirable 11-25 mg l(-1) in the final effluent. The levels of methane-producing activity from acetate and H2/CO2 gas at 10 degrees C were only 2% and 0% of those at 35 degrees C, respectively. On the other hand, the sulfate-reducing activity levels of the UASB sludge were relatively high at 10 degrees C, for example, 18% for acetate and 9% for H2/CO2 gas, compared to the activity levels at 35 degrees C. Therefore, BOD oxidization by sulfate reduction in the UASB was greater than that by methane production under low temperature conditions. This sulfate-reducing activity tended to be proportional to the copy number of adenosine-5'-phosphosulfate (APS) reductase genes in DNA extracted from the sludge.