Genotoxicity biomonitoring of sewage in two municipal wastewater treatment plants using the Tradescantia pallida var. purpurea bioassay

The genotoxicity of untreated and treated sewage from two municipal wastewater treatment plants (WTP BN and WTP SJN) in the municipality of Porto Alegre, in the southern Brazilian state of Rio Grande do Sul, was evaluated over a one-year period using the Tradescantia pallida var. purpurea (Trad-MCN) bioassay. Inflorescences of T. pallida var. purpurea were exposed to sewage samples in February (summer), April (autumn), July (winter) and October (spring) 2009, and the micronuclei (MCN) frequencies were estimated in each period. The high genotoxicity of untreated sewage from WTP BN in February and April was not observed in treated sewage, indicating the efficiency of treatment at this WTP. However, untreated and treated sewage samples from WTP SJN had high MCN frequencies, except in October, when rainfall may have been responsible for reducing these frequencies at both WTPs. Physicochemical analyses of sewage from both WTPs indicated elevated concentrations of organic matter that were higher at WTP SJN than at WTP BN. Chromium was detected in untreated and treated sewage from WTP SJN, but not in treated sewage from WTP BN. Lead was found in all untreated sewage samples from WTP SJN, but only in the summer and autumn at WTP BN. These results indicate that the short-term Trad-MCN genotoxicity assay may be useful for regular monitoring of municipal WTPs.     

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.     

Effects of Sewage Sludge on the Growth of Potted Salt Marsh Plants Exposed to Natural Tidal Inundation

Abstract A growth experiment with native plants in pots exposed to natural environmental conditions evaluates the use of sewage sludge as a soil amendment in restoration of a southern California salt marsh. Sludge containing desirable organic matter but also undesirable heavy metal contaminants was mixed with a readily available matrix soil to reduce metal concentrations to levels below legal limits for land applications of sludge. Soil nutrient analysis revealed expected increases in total nitrogen and total phosphorus content with increasing sludge concentration. Soil metals analysis, however, revealed decreases in metal content with increased sludge concentration, a trend evidently caused by higher than expected metal content in the matrix soil. Five artificial soil mixtures ranging from 0 to 70% sludge were accompanied by natural wetland soil controls. Pots containing these soils were placed into a natural salt marsh. The pots were then planted with two native salt marsh plant species, Salicornia virginica and Frankenia grandifolia. Aboveground biomass was harvested after 12 months. Plant growth displayed no obvious change with increasing sludge concentration. Over the concentration ranges used, increased nutrient content did not stimulate plant growth and increased metal content did not inhibit plant growth. Plants grew better in natural wetland soil than in artificial soil mixtures, a trend probably caused by the substantially finer texture and higher organic content of natural soil. All sludge treatments differed more from the natural soil than from each other, implying that within the ranges examined, soil texture and organic content exerted more influence on plant growth than did metal or nutrient concentration. These results suggest that incorporating this sewage sludge in the soil of the restored salt marsh will neither benefit nor harm the plants that will live there and that greatest plant growth will be achieved by mixing the sludge with a fine‐grained matrix soil.     

几种花卉植物对污泥中铅的富集特征

土地利用是资源化利用城市污水厂污泥的有效途径。为了解污泥中重金属污染物在土壤、植物之间的迁移富集规律,选用8种园林用植物进行污泥混配土种植实验。结果表明,植物对污泥混配土土壤中铅的富集能力,以及铅在植物中各器官的富集系数均不同,且不同种类植物差别较大。植物根部对铅有较强的富集能力,其次是叶和茎。菊花对铅的富集能力较强。植物对土壤中铅的富集能力也受土壤铅含量和赋存形态影响。     

表施和混施污泥对团花根系生长的影响

林地利用被认为是污泥资源化利用的重要方式,但施用污泥对林木根系生长的影响报道较少。本研究通过根箱试验,分析表施和混施10%污泥对速生树种团花不同土层根系形态、土壤pH值和电导率动态变化及根系重金属含量的影响,并拟合土壤pH值、电导率和根系重金属含量与根长的关系。结果表明: 与不施污泥(对照)相比,混施污泥显著抑制了团花根长、根表面积和根体积增长,混施污泥120和240 d后,0～20 cm土层总根长分别为不施污泥的76.9%和67.4%;表施污泥对团花根长和根表面积的影响不显著,但显著提高了根体积。混施污泥显著提高了土壤pH值和电导率及根系重金属含量,混施污泥0～20和20～40 cm土层根系镉含量分别是不施污泥的11.5和10.0倍。线性回归拟合分析显示,不同处理0～20 cm土层的电导率与根长均呈显著负相关;表施和混施污泥根系镉含量与根长呈极显著负相关。上述结果表明,混施污泥抑制了团花根系生长,这可能是由于混施污泥提高了土壤电导率和根系镉含量所致,而表施污泥对团花根系生长的作用不明显。     

Ecophysiological Responses of Plants After Sewage Sludge Compost Applications

Composting is one of the most appropriate methods to recycle sewage sludge. Sewage sludge compost is a suitable solution for improving the quality of barren soil at landfill. Therefore, it is important to investigate the effects of sewage sludge compost on plants. Different compost application methods (mixing and scattering over reclaimed soil) on sawtooth oak (Quercus acutissima) and Japanese red pine (Pinus densiflora) have been tested. The application of sewage sludge compost markedly increased soil moisture and nitrogen content. Compost treatments resulted in significant increases in both plant height and biomass as compared to controls. Compost treatments led to a significant increase in the N content of plant leaves. Compost treatments resulted in significant increases in the chlorophyll content and photosynthetic rates of the plants. The scattering of compost over reclaimed soil (compost 2) resulted in lower total antioxidant activity and superoxide dismutase activity than mixing the compost with the reclaimed soil (compost 1), or in the control treatment. Since the growth rates, N content, and photosynthetic rates in compost 2 treatment were not markedly different from compost 1 treatment, it (compost 2) would be a better application method from both an ecological and economic perspective.     

Effects of nickel hyperaccumulation on physiological characteristics of Alyssum murale grown on metal contaminated waste amended soil

A pot experiment was conducted to investigate the effect of nickel concentration on physiological characteristics of Alyssum murale when grown in a soil mixed with sewage sludge (at the rate of 2.8%). Two types of sludge were used: agricultural sewage sludge (S1) and industrial sewage sludge with an increasing nickel concentration (S2, S3, and S4). Results showed that Ni in shoots was higher than Ni in roots. A. murale is able to concentrate up to 12730 mg/kg Ni in leaves. The highest dry matter yield was observed with plants grown with agricultural sewage sludge. An addition of S2 and S3 increased shoot biomass. However, application of S4 reduced 40% shoot dry weight as compared to the control Ni treatment did not affect all chlorophyll fluorescence parameters. The F(v)/F(m) ratio was stable between Ni treatments. Photosynthesis rate (A) increased with agricultural sewage sludge, but remained stable with variable Ni rates from the industrial sludge. The chlorophyll content increased with S1, S2 and S3 but it remains constant with S4 when compared to the control Therefore, high nickel concentration did not affect the function of the photosynthetic machine of A. murale.     

Occurrence and activity of Archaea in aerated activated sludge wastewater treatment plants

The occurrence, distribution and activity of archaeal populations within two aerated, activated sludge wastewater treatment systems, one treating domestic waste and the second treating mixed domestic and industrial wastewater, were investigated by denaturing gradient gel electrophoresis (DGGE) analysis of polymerase chain reaction (PCR)-amplified ribosomal RNA gene fragments and process measurements. In the plant receiving mixed industrial and domestic waste the archaeal populations found in the mixed liquor were very similar to those in the influent sewage, though a small number of DGGE bands specific to the mixed liquor were identified. In contrast, the activated sludge treating principally domestic waste harboured distinct archaeal populations associated with the mixed liquor that were not prevalent in the influent sewage. We deduce that the Archaea in the plant treating mixed wastewater were derived principally from the influent, whereas those in the plant treating solely domestic waste were actively growing in the treatment plant. Archaeal 16S rRNA gene sequences related to the Methanosarcinales, Methanomicrobiales and the Methanobacteriales were detected. Methanogenesis was measured in activated sludge samples incubated under oxic and anoxic conditions, demonstrating that the methanogens present in both activated sludge plants were active only in anoxic incubations. The relatively low rates of methanogenesis measured indicated that, although active, the methanogens play a minor role in carbon turnover in activated sludge.     

Simultaneous detection of Cryptosporidium parvum and Giardia in sewage sludge by IC-PCR

AIMS: The aim of this study was to develop a method based on immunomagnetic capture and polymerase chain reaction (IC-PCR assay) for detection of Cryptosporidium parvum and Giardia intestinalis in sewage sludge. METHODS AND RESULTS: The detection limit of the IC-PCR assay for both organisms was 625 oocysts and cysts ml(-1). By hybridization of PCR products the sensitivity could be increased to 125 oocysts and cysts ml(-1). Forty-four sludge samples from 12 wastewater treatment plants were examined. The samples positive for Giardia (9 out of 44) were from eight wastewater plants and the C. parvum genotype 2 samples (3 out of 44) originated from different sewage works. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: IC-PCR offers the possibility to distinguish between Cryptosporidium and Giardia genotypes. This assay can be used to monitor the presence of these organisms in a community and to determine contamination of sludge used as soil amendment.     

Near-Bottom Pelagic Bacteria at a Deep-Water Sewage Sludge Disposal Site

The epibenthic bacterial community at deep-ocean sewage sludge disposal site DWD-106, located approximately 106 miles (ca. 196 km) off the coast of New Jersey, was assessed for changes associated with the introduction of large amounts of sewage sludge. Mixed cultures and bacterial isolates obtained from water overlying sediment core samples collected at the deep-water (2,500 m) municipal sewage disposal site were tested for the ability to grow under in situ conditions of temperature and pressure. The responses of cultures collected at a DWD-106 station heavily impacted by sewage sludge were compared with those of samples collected from a station at the same depth which was not contaminated by sewage sludge. Significant differences were observed in the ability of mixed bacterial cultures and isolates from the two sites to grow under deep-sea pressure and temperature conditions. The levels of sludge contamination were established by enumerating Clostridium perfringens, a sewage indicator bacterium, in sediment samples from the two sites. The results of hybridization experiments in which DNAs extracted directly from the water overlying sediment core samples were used indicate that the reference site epibenthic community, the disposal site epibenthic community, and the community in a surface sludge plume share many members. Decreased culturability of reference site mixed cultures in the presence of sewage sludge was observed. Thus, the culturable portions of both the autochthonous and allochthonous bacterial communities at the disposal site may be inhibited in situ, the former by sewage sludge and the latter by high pressure and low temperature.     

Influence of soil type, moisture content and biosolids application on the fate of Escherichia coli in agricultural soil under controlled laboratory conditions

AIMS: To determine the fate of the enteric indicator organism, Escherichia coli, in sewage sludge (biosolids)-amended agricultural soil in relation to soil type and moisture status under controlled conditions. METHODS AND RESULTS: We enumerated Escherichia coli in soil by membrane filtration and most probable number techniques. The background concentration of E. coli was higher in sandy loam than in silty clay soil. E. coli numbers increased in soil following addition of dewatered, mesophilic anaerobically digested sludge. Escherichia coli declined to a small extent with time in both moist and air-dried unamended control soils, although decay was only highly significant (P < 0.001) in moist sandy loam (T(90) = 100 days). Removal rates were high in sludge-treated moist soil (T(90) = 20 days), but were significantly reduced in amended air-dried soil. CONCLUSIONS: Slow removal of E. coli in air-dried soil as against their rapid decay in moist soil after sludge application indicated that the soil biota are involved in pathogen reduction processes in sludge-amended soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Soil ecological mechanisms are implicated as having a critical role in the fate of enteric organisms introduced into temperate agricultural soil in sewage sludge.     

Agricultural reclamation of disturbed soils in a lignite mining area using municipal and coal wastes: the humus situation at the beginning of reclamation

Soils disturbed by long-term opencast mining were treated with organic waste materials for reclamation. Humic substances were extracted from waste and soil samples and analysed using pyrolysis-gas chromatography/mass spectrometry and electrofocusing. Furthermore, analytical pyrolysis permits to study all starting materials in situ. According to structural similarities, the statistical evaluation of the pyrolysis results clearly indicates three sample groups. The first group, called compost, implies the waste materials compost and composted sewage sludge. Moreover, pyrolysis revealed that coal humic substances are predominant in brown coal sludge, pure mine soils and mine soils treated with the different organic waste materials. They constitute the second group. The sewage sludge contains a high nitrogen potential, as expected, and represents the third group. Finally, pyrolysis generally showed the specific structural characteristics of humic and fulvic acids, respectively. Electrofocusing yielded for all samples a signal pattern that is typical of humic substances. However, number and ratio of the signals differ according to the special structural features of the samples. This revised version was published online in June 2006 with corrections to the Cover Date. This revised version was published online in June 2006 with corrections to the Cover Date. This revised version was published online in June 2006 with corrections to the Cover Date.     

Monitoring the genotoxicity of soil extracts from two heavily polluted sites in Prague using the Tradescantia stamen hair and micronucleus (MNC) assays.

We have taken soil samples from two sites in Prague, the capital of the Czech Republic, that are heavily polluted by motor vehicles. As a negative control, soil samples from a recreational site in Prague with no motor vehicle traffic were used. Soil samples from these sites were extracted with water or 5% dimethylsulphoxide (DMSO) for 24 h and cuttings of Tradescantia clone 4430 were immersed for 12 h at 25 degrees C in the extraction solutions. As a positive control Tradescantia plants have been treated with the promutagenic arylamine o-phenylenediamine at the same treatment conditions. None of the tested soil extractions significantly increased the frequency of somatic mutations in the stamen hair assay. By contrast, a 5% DMSO soil extract from one of the tested sites (entrance of the Letná tunnel) significantly increased the frequency of micronuclei (MNC) in the pollen mother tetrad cells. A repetition of the treatment 14 days later also resulted in an increase in the frequency of MNC, however the increase was not statistically significant. This study was conducted for the International Programme on Plant Bioassays.     

Toxic effects of digested, composted and thermally-dried sewage sludge on three plants

In order to evaluate potential toxic effects of stabilized sewage sludge that are currently applied to agricultural soils, three types of municipal sewage sludge and one pig slurry were subjected to phytotoxicity assays using three plants (Brassica rapa, Lolium perenne and Trifolium pratense). Equivalent batches of aerobically and anaerobically-digested sludge (F) from two municipal wastewater treatment plants, were composted (C) or thermally dried (T). In addition, one anaerobically-digested and thermally-dried pig slurry (P) was tested. A seedling growth test was performed in accordance with the OECD Guideline 208A, using seed emergence and shoot length as endpoints to identify the inhibition of plants growing in increasing doses of sludge. A correlation analysis between EC50 and physico-chemical parameters or the pollutant burden of the biosolids was also calculated. In all tests, lower germination rates were observed for T. pratense than for L. perenne and B. rapa. A moderate stimulatory effect on shoot length at low doses was observed for the three plants. In these wastes, a strong positive correlation was found between higher values of EC50 (less toxicity) and the stability degree of their organic matter, and a negative correlation between EC50 and total N, hydrolysable N or NH4-N content. No correlations were found with heavy metal or organic pollutant content in those wastes. Results indicate that digested sludge, thermally-dried sludge and thermally-dried pig slurry have non-negligible short term phytotoxic effects, and confirm that composting is an effective sludge treatment for the reduction of phytotoxicity.     

Effects of Municipal Sewage Sludge Doses on the Chlorophyll Contents and Heavy Metal Concentration of Sugar Beet (Beta vulgaris var. saccharifera)

The present study was conducted to assess the suitability of sewage sludge amendment (SSA) in soil for Beta vulgaris var. saccharifera (sugar beet) by evaluating the heavy metal accumulation and physiological responses of plants grown at a 10%, 25%, and 50% sewage sludge amendment rate. The sewage sludge amendment was modified by the physicochemical properties of soil, thus increasing the availability of heavy metals in the soil and consequently increasing accumulation in plant parts. Cd, Pb, Ni, and Cu concentrations in roots were significantly higher in plants grown at 25% as compared to 50% SSA; however, Cr and Zn concentration was higher at 50% than 25% SSA. The concentrations of heavy metal showed a trend of Zn > Ni > Cu > Cr > Pb > Cd in roots and Zn > Cu > Ni > Cr > Pb > Cd in leaves. The only instance in which the chlorophyll content did not increase after the sewage sludge treatments was 50%. There were approximately 1.12-fold differences between the control and 50% sewage sludge application for chlorophyll content. The sewage sludge amendment led to a significant increase in Pb, Cr, Cd, Cu, Zn, and Ni concentrations of the soil. The heavy metal accumulation in the soil after the treatments did not exceed the limits for the land application of sewage sludge recommended by the US Environmental Protection Agency (US EPA). The increased concentration of heavy metals in the soil due to the sewage sludge amendment led to increases in heavy metal uptake and the leaf and root concentrations of Ni, Zn, Cd, Cu, Cr, Pb, and Zn in plants as compared to those grown on unamended soil. More accumulation occurred in roots and leaves than in shoots for most of the heavy metals. The concentrations of Cd, Cr, and Pb were more than the permissible limits of national standards in the edible portion of sugar beet grown on different sewage sludge amendment ratios. The study concludes that the sewage sludge amendment in the soil for growing sugar beet may not be a good option due to risk of contamination of Cr, Pb, and Cd.     

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。     

Effects of different soil ameliorants on karee trees (Searsia lancea) growing on mine tailings dump soil-part I: pot trials

Rehabilitation of mine tailings dams is often a challenge due to a lack of nutrients and a poor humus reservoir prevailing in tailings soils. This is especially true for establishing longer lived species such as trees. For these reasons the effects of different soil ameliorants (woodchips compost, vermicompost, mature sewage sludge), added to the root system of Karee (Searsia lancea) saplings were tested in pot trials. Those pots were filled with platinum and gold tailings substrate as well as red clay soil, respectively. For three months plants remained in a greenhouse and were subsequently moved to a test field outside. Throughout the test period regular chl a fluorescence measurements were taken and subjected to JIP-test quantifying changes in photosynthetic vitality status. Additionally, growth measurements and one-off leaf analysis were carried out. Test plants growing on mine tailings experienced an up to 35% higher average photosynthetic vitality (PI(ABS)) and improved nutrient supply, when treated with mature sewage sludge. Consequently, sewage sludge treated plants showed a higher biomass build-up rate and an up to 55% higher diameter growth, compared to control. In summary the experiments present a low cost alternative for reforestation enterprises on platinum and gold tailings dams in South Africa.     

Efficacy of Biosolids in Assisted Phytostabilization of Metalliferous Acidic Sandy Soils with Five Grass Species

The role of sewage sludge as an immobilising agent in the phytostabilization of metal-contaminated soil was evaluated using five grass species viz., Dactylis glomerata L., Festuca arundinacea Schreb., F. rubra L., Lolium perenne L., L. westerwoldicum L. The function of metal immobilization was investigated by monitoring pH, Eh and Cd, Pb, and Zn levels in column experiment over a period of 5-months. Grasses grown on sewage sludge-amendments produced high biomass in comparison to controls. A significant reduction in metal uptake by plants was also observed as a result of sewage sludge application, which was attributed to decreased bioavailability through soil stabilisation. We have observed that the sludge amendment decreased metal bioavailability and concentrations in soil at a depth of 25 cm, in contrast to untreated columns, where metal concentrations in the soil solution were very high.     

The use of sewage sludge and horticultural waste to develop artificial soil for plant cultivation in Singapore

Greenhouse pot experiments were performed with Ipomoea aquatica (Kang Kong) to evaluate artificial soil produced from poor fertility subsoil, horticultural compost, and sewage sludge. The addition of horticultural compost and sewage sludge to subsoil substantially improved plant growth, improved the physical properties of subsoil and enriched subsoil by essential nutrients for plants. The effect was enhanced when the two ingredients were added to subsoil together. The highest yield of biomass of I. aquatica was observed in artificial soil prepared by mixing subsoil with 4% (wet weight/wet weight) of horticultural compost and 2% (dry weight/wet weight) of sewage sludge. The contents of heavy metals in plants, grown in the artificial soil, were significantly lower than toxic levels. The artificial soil could be recommended for urban landscaping and gardening in Singapore.     

Impact of application of organic waste materials on microbial and enzyme activities of mine soils in the Lusatian coal mining region

The objective of the present work was to study the short-term stimulation of microbial and enzyme activity in mine soils by application of organic waste materials in lysimeter and mesocosm studies. The mine soils derived from tertiary and quaternary deposits were ameliorated with brown coal filter ash (tertiary deposits) and lime (quaternary deposits). At the beginning of recultivation the soils were treated with varying amounts of sewage sludge, coal sludge, composted sewage sludge and compost to a depth of 30 cm. In the first 2 years after application of organic waste materials we found a very low level of microbial properties especially in the sandy materials from quaternary deposits but a significant increase in microbial respiration, substrate induced respiration and enzyme activities like invertase and alkaline phosphatase with increasing application rates of sewage sludge, compost and sewage sludge mixed with coal sludge. This can be explained by an increase in organic matter and nutrient content of the soils and an improvement of soil physical properties such as water and nutrient retention capacity. Additionally it can be assumed, that constituents of the coal admixtures of tertiary deposits can be mineralised or converted by the soil microorganisms. In the tertiary materials ameliorated with brown coal ash the highest amounts of microbial and enzyme activities were measured after application of nitrogen-rich sewage sludge or very high amounts of mature compost mainly consisting of green waste. Compared with sewage sludge the stimulating effects of composted sewage sludge were quite lower because of organic matter fragmentation and a reduced energy and nutrient supply to soil microorganisms. This revised version was published online in June 2006 with corrections to the Cover Date.