Bacterial diversity in thermophilic aerobic sewage sludge

Summary Using -amylase as an example, extremely thermophilic Bacilli isolated from heat-treated sewage sludge are shown to be a source for enzymes stable and active at high temperatures. The isolates which are classified as subspecies of Bacillus stearothermophilus differ from each other in protein composition indicating the heterogeneiety of that subspecies. Media are evaluated for good growth and high enzyme productivity. Best media are those composed of three or four different complex components like combinations of peptone, soy grist, and malt extract, -amylase production on simple carbon sources is negligible. From the cultivation supernatants crude -amylase extracts are prepared and their behaviour at high temperatures is described. The optimal temperature of all tested enzymes is 80°C. They are stable at suboptimal temperatures for over 20 h and at 95° C 50% of their activity is lost within 2 h. The activity at 95° C is however preserved for over 3 h in presence of starch. The products of the starch digestion are maltotriose, maltose, and some glucose. The amylases can therefore compete in activity and stability with commercially available -amylases from Bacillus licheniformis.     

Leaching of iron and toxic heavy metals from anaerobically-digested sewage sludge

Summary Heavy metal-loaded sewage sludge was leached abiotically using FeCl₂ and FeCl₃ which are applied in waste water treatment plants to eliminate phosphate and for coagulation. Due to the hydrolyzing nature of ferric iron, ferric chloride (100 mmll L^–1) was able to solubilize more than 90% of copper and zinc and more than 80% of cadmium, with an optimal pulp density of 3% (w/v), after 10 h of exposition at 25°C. Chromium, lead and nickel were solubilized to an extent of 40–70%. With the exception of copper (redoxolysis), all heavy metals monitored were leached following the principle of acidolysis. Chemical leaching with iron resulted in a secondary contamination of sewage sludge (96 g iron per kg dry weight). The insoluble iron compounds which were precipitated for adsorbed to sludge flocks could be resolubilized with oxalic acid (100 mM, pH<3.3) up to an extent of 90%. Iron was leached by acidolysis and held in solution by complexation with oxalic acid. The pH optimum for the treatment of sewage sludge with 100 mmol L^–1 oxalic acid was pH 3.3. At this pH an excessive solubilization of nutrient elements and compounds (phosphorus, nitrogen, alkali and alkali earth elements) could be avoided concomitantly leaching 75% iron. Furthermore the hydrophobicity of the sewage sludge was significantly reduced as a result of treatment with iron chloride.Thiobacillus ferrooxidans (isolated from arsenopyrite and adapted on sewage sludge) utilized ferrous iron as an energy source in the presence of chloride ions (FeCl₂) as efficiently as ferrous sulphate. No toxic effects of oxalic acid onT. ferrooxidans were observed at the prevailing concentration.     

Effects of sewage sludge application on heavy metal leaching from mine tailings impoundments

Column experiments were conducted to investigate the removal of heavy metals from two mine tailings (El Arteal and Jaravías) using sewage sludge as a reactive material. When sewage sludge is used as a reactive material on the El Arteal tailings (sample SA), Fe, Mn, Zn and Pb are removed and Cu and Ni are mobilized. The experiments carried out on the Jaravías tailings give similar results, showing the retention of Cu, Pb, Fe and Mn and the mobilization of Ni and Zn. An analysis performed using the PHREEQC numerical code suggests that the retention of Fe in the sewage sludge may be caused by the precipitation of Fe(OH)2.7Cl0.3 and possibly pyrite, and that the retention of Pb at high pH may be caused by the formation of stable phase minerals such as Pb(OH)2 and PbS in these conditions. Ni mobilization in the column experiments with the two tailings samples may be caused by the presence of significant amounts of leachable Ni in the sewage sludge. The complexation of metals with dissolved organic matter, calculated with the Minteq model, may be moderate.     

Influence of thermal treatment on sequential extraction and leaching behaviour of trace metals in a contaminated sewage sludge

The losses of weight and organic matter of a sludge caused by thermal treatments at 180 degrees C, 300 degrees C and 400 degrees C were determined in order to assess how the possibilities of sludge use were influenced. The sludge heated at 180 degrees C lost small amounts of weight and organic matter (9.8%) but the losses from the two other treatments were large enough (92.2% and 99.9% in organic matter) to preclude the use of the sludges as organic amendments. The concentration and potential lability and leachability of Cr, Cu, Fe, Mn, Ni, Pb and Zn in the native sludge and in the thermal-treated sludge samples were studied by means of a five-step chemical fractionation method and a column experiment. As a consequence of heating, the trace metals were more strongly fixed in the treated sludges, as could be seen by the decrease with temperature of the ratio between the sum of the first two sequential-extracted fractions and the residual fraction. The leaching results showed that, for the native sludge, the quantities of studied metals leached were larger than for the sludge heated to 180 degrees C. The order of leachability of metals was the same in both cases, and the same equation could be used to calculate the quantities of metals leached. The amounts of metals leached correlated significantly with the first extracted fraction (exchangeable metals) and an equation could be used to calculate the quantities leached, as a function of that fraction.     

Thermophilic microbial leaching of heavy metals from municipal sludge using indigenous sulphur-oxidizing microbiota

 It was demonstrated in shake-flask experiments that the sulphur-oxidizing microbiota of municipal sludges can be used at 53°C for heavy-metal leaching. Five sludges were tested and the average final pH, oxidation/reduction potential and SO^2- ₄ concentration after 30 days were 2.8, 237 mV and 5668 mg/l respectively. Ferric chloride was added to enhance the redox potential and to lower pH, which resulted in average values of 409 mV and 1.86 respectively. The average solubilisation of Cd, Cu, Cr, Mn, Ni, Pb, Zn, K and P after ferric chloride addition was 35.6±20.6%, 65.9±15.1%, 28.5±13.5%, 74.0±10.0%, 60.3± 13.1%, 33.7±27.6%, 83.9±6.2%, 39.0±16.6 and 18.2±15.8% respectively. The present process enhanced the sludge dewaterability compared to the conventional thermophilic digestion. During the leaching batch process, the volatile and volatile suspended solids were degraded to the same level as observed when the conventional thermophilic digestion was used as control. Received: 7 June 1995/Received revision: 8 September 1995/Accepted: 29 September 1995     

Phytoextraction of heavy metals from municipal sewage sludge by Rosa multiflora and Sida hermaphrodita

The aim of the study was to evaluate the efficacy of the multiflora rose var. “Jatar” (Rosa multiflora Thunb. ex Murray) and the Virginia fanpetals (Sida hermaphrodita Rusby) to phytoextract heavy metals from municipal sewage sludge. The 6-year field experiment involved four levels of fertilization with sewage sludge at doses of 0, 10, 20, 40, and 60 Mg DM (Dry Mass) sludge ha^?1. The increasing doses of sewage sludge were found to significantly increase the yield of multiflora rose and Virginia fanpetals biomass. They also significantly increased the content of heavy metals in these plants. The highest uptake of heavy metals by the multiflora rose and Virginia fanpetals crops was recorded at the fertilization dose of 60 Mg DM ? ha^?1. Our investigations show that the Virginia fanpetals was more efficient in the phytoextraction of Cr, Ni, Cu, Zn, and Cd from the sewage sludge than the multiflora rose, due to the greater yields and higher heavy metal uptake by the former plant. In turn, the multiflora rose phytoextracted greater amounts of Pb from the sewage sludge. The analyses indicate that the Virginia fanpetals can be used for phytoremediation (phytoextraction) of heavy metals contained in sewage sludge.     

Immobilization of selected heavy metals in sewage sludge by natural zeolites

This contribution presents the possibility of application of natural sorbent (Transcarpathian clinoptylolite (KL)) for immobilization of selected heavy metals in the sewage sludge. The influence of ion-exchange parameters (e.g. time, amount of zeolite) were discussed. Process of immobilization was performed using a static method (Batch). It was found that best possible conditions for immobilization of heavy metal ions were as follows: zeolite fraction 0.7–1.0 mm, 5 h of shaking, zeolite/sewage sludge ratio 2/98.     

Removal of heavy metals from contaminated sewage sludge using Aspergillus niger fermented raw liquid from pineapple wastes

The environmental benefits derived from using citric acid in the removal of heavy metals from contaminated sewage sludge have made it promising as an extracting agent in the chemical extraction process. At present, citric acid is produced commercially by fermentation of sucrose using mutant strains of Aspergillus niger (A. niger), and chemical synthesis. In recent years, various carbohydrates and wastes (such as pineapple wastes) have been considered experimentally, to produce citric acid by A. niger. This study investigated the potential of using A. niger fermented raw liquid from pineapple wastes as a source of citric acid, in extracting chromium (Cr), copper (Cu), lead (Pb), nickel (Ni) and zinc (Zn) from anaerobically digested sewage sludge. Results of the study revealed that metal removal efficiencies varied with pH, forms of metals in sludge and contact time. At pH approaching 4, and contact time of 11 days, A. niger fermented liquid seemed to remove all Cr and Zn while removing 94% of Ni. Moreover, chemical speciation studies revealed that metals which are predominantly in the exchangeable and oxidizable phases seemed to exhibit ease of leachability (e.g., Zn). The by-products of the process such as pineapple pulp and mycelium which are rich in protein, can still be used as animal feed. It can be said therefore that this novel process provides a sustainable way of managing contaminated sewage sludge.     

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).     

Bacterial removal of mercury from sewage

Mercury-resistant bacteria, which are able to reduce mercuric ion (Hg(2+)) to metallic mercury (Hg(0)), were examined for their ability to remove mercury from waste-water aerobically. Growth studies in artificial medium indicated that mercury increases the lag phase, but does not effect the growth rate of these bacteria. Further studies demonstrated that growth was minimal during a phase of rapid mercury removal, after which growth resumed. Small but significant amounts of carbohydrates are required for the mercuric ion reduction. Prolonged periods of bacterial growth under nonsterile conditions was accomplished without the loss of the mercuric reducing ability of the culture. A continuous culture of the resistant organism was maintained on raw sewage for two weeks, during which time relatively high concentrations of mercury (70 mg/L) were removed from the sewage at a rate of 2.5 mg/L h and at efficiencies exceeding 98%.     

Microbial leaching of metals from sulfide minerals

Microorganisms are important in metal recovery from ores, particularly sulfide ores. Copper, zinc, gold, etc. can be recovered from sulfide ores by microbial leaching. Mineral solubilization is achieved both by 'direct (contact) leaching' by bacteria and by 'indirect leaching' by ferric iron (Fe(3+)) that is regenerated from ferrous iron (Fe(2+)) by bacterial oxidation. Thiobacillus ferrooxidans is the most studied organism in microbial leaching, but other iron- or sulfide/sulfur-oxidizing bacteria as well as archaea are potential microbial agents for metal leaching at high temperature or low pH environment. Oxidation of iron or sulfur can be selectively controlled leading to solubilization of desired metals leaving undesired metals (e.g., Fe) behind. Microbial contribution is obvious even in electrochemistry of galvanic interactions between minerals.     

施用污泥对小白菜生长及其迁转重金属的影响

采用盆栽方式,以厦门市筼筜污水处理厂的污泥作为有机肥种植小白菜,探讨污泥施用对小白菜生长和产量的影响及重金属在小白菜-土壤间的迁移转化和积累情况.结果表明:污泥中氮、磷、钾和有机质含量丰富,重金属含量均低于国家农用污泥中污染物控制标准;适宜的污泥施用量可促进小白菜的生长,污泥用量为5%时,小白菜长势最好,生物量最大;污...     

Long-term effects of metals in sewage sludge on soils,microorganisms and plants

Summary This paper reviews the evidence for impacts of metals on the growth of selected plants and on the effects of metals on soil microbial activity and soil fertility in the long-term. Less is known about adverse long-term effects of metals on soil microorganisms than on crop yields and metal uptake. This is not surprising, since the effects of metals added to soils in sewage sludge are difficult to assess, and few long-term experiments exist. Controlled field experiments with sewage sludges exist in the UK, Sweden, Germany and the USA and the data presented here are from these long-term field experiments only. Microbial activity and populations of cyanobacteria,Rhizobium leguminosarum bv.trifolii, mycorrhizae and the total microbial biomass have been adversely affected by metal concentrations which, in some cases, are below the European Community's maximum allowable concentration limits for metals in sludge-treated soils. For example, N₂-fixation by free living heterotrophic bacteria was found to be inhibited at soil metal concentrations of (mg kg^–1): 127 Zn, 37 Cu, 21 Ni, 3.4 Cd, 52 Cr and 71 Pb. N₂-fixation by free-living cyanobacteria was reduced by 50% at metal concentrations of (mg kg^–1): 114 Zn, 33 Cu, 17 Ni, 2.9 Cd, 80 Cr and 40 Pb.Rhizobium leguminosarum bv.trifolii numbers decreased by several orders of magnitude at soil metal concentrations of (mg kg^–1): 130–200 Zn, 27–48 Cu, 11–15 Ni, and 0.8–1.0 Cd. Soil texture and pH were found to influence the concentrations at which toxicity occurred to both microorganisms and plants. Higher pH, and increased contents of clay and organic carbon reduced metal toxicity considerably. The evidence suggests that adverse effects on soil microbial parameters were generally found at surpringly modest concentrations of metals in soils. It is concluded that prevention of adverse effects on soil microbial processes and ultimately soil fertility, should be a factor which influences soil protection legislation.     

Metaproteome analysis of sewage sludge from membrane bioreactors

Microbial dynamics and enzymatic activities of activated sludge processes are not completely understood yet. A better understanding about the biology is indispensable for further process optimization. Since proteins play a key role as catalysts in sludge processes, a protocol for protein extraction and analysis by 2-D PAGE was established. It is based on phenol extraction of alkaline extracts and on a subsequent precipitation with ammonium sulphate. 2-D protein patterns obtained from different sludges collected from membrane bioreactors showed--besides common spots--significant differences. Selected proteins were identified with nano-HPLC-ESI-MS/MS. All membrane biological reactor (MBR) sludge samples investigated in this study contained elastase 3A, which implies that this human serine protease is a significant constituent of municipal wastewater. Although the identification of proteins from ammonia-oxidizing bacterium Nitrosomonas europaea was expected, the detection of a protein with homology to the marine bacterium Saprospira grandis in MBR1 was surprising.     

Microbial leaching of metals from solid industrial wastes

Biotechnological applications for metal recovery have played a greater role in recovery of valuable metals from low grade sulfide minerals from the beginning of the middle era till the end of the twentieth century. With depletion of ore/minerals and implementation of stricter environmental rules, microbiological applications for metal recovery have been shifted towards solid industrial wastes. Due to certain restrictions in conventional processes, use of microbes has garnered increased attention. The process is environmentally-friendly, economical and cost-effective. The major microorganisms in recovery of heavy metals are acidophiles that thrive at acidic pH ranging from 2.0–4.0. These microbes aid in dissolving metals by secreting inorganic and organic acids into aqueous media. Some of the well-known acidophilic bacteria such as Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, Leptospirillum ferrooxidans and Sulfolobus spp. are well-studied for bioleaching activity, whereas, fungal species like Penicillium spp. and Aspergillus niger have been thoroughly studied for the same process. This mini-review focuses on the acidophilic microbial diversity and application of those microorganisms toward solid industrial wastes.     

Total concentrations and chemical speciation of heavy metals in liquefaction residues of sewage sludge

The risk (including bioavailability and eco-toxicity) of heavy metals (Pb, Zn, Cu, Cd, Cr and Ni) in liquefaction residues (LR) of sewage sludge (SS) was estimated, according to both the speciation of heavy metals and the local environmental characteristics. The amount of organic matters in LR was lower than that in SS, resulting in a smaller calorific value, while the total content of heavy metals in LR nearly doubled. High residual rates of heavy metals (about 80%) indicated that the heavy metals in SS were concentrated into LR after liquefaction. The comparisons of sequential extraction results between SS and LR showed that after liquefaction, the mobile and easily available heavy metal fractions (acid soluble/exchangeable and reducible fractions) were mainly transformed into the relatively stable heavy metal fractions (oxidizable and residual fractions). The bioavailability and eco-toxicity of heavy metals in LR were relieved, though the total concentrations of heavy metals increased.     

施污土壤重金属有效态分布及生物有效性

以城市污泥为研究对象,将城市污泥与土壤按照一定的质量比配成污泥混合土壤.采用6种不同性质提取剂(0.05mol/L EDTA、0.1 mol/L CH3 COOH、0.01 mol/L CaC12、1 mol/L CH3COONH4,0.05 mol/L NaHCO3和0.05 mol/L Tris-HCl)分别对污泥混合土壤中重金属(Cd、Pb、Cu和Zn)的螯合态、酸溶态、中性交换态、中性结合态、碱性交换态和蛋白质结合态进行提取,考察污泥的添加对土壤中不同形态重金属的消长规律.通过黑麦草盆栽试验,探究污泥混合土壤中不同形态重金属的植物可利用性.结果表明:污泥混合土壤中重金属螯合态比例较大,占总量的20.3％-40.0％;其次为酸溶态和中性结合态,而中性交换态、碱性交换态和蛋白质结合态的含量较低.污泥的添加促进了黑麦草对Cd、Cu和Zn的吸收,在污泥添加率为44.4％时根部对其吸收量达最大,分别较CK处理增加了0.3、2.3和6.5倍.抑制了对Pb的吸收,在污泥添加率为37.5％时,根部对Pb的吸收较CK处理下降0.4倍.Pearson相关系数分析结果表明:污泥混合土壤中以螯合态、酸溶态和中性结合态存在重金属可被黑麦草吸收利用.     

Foam control in biopesticide production from sewage sludge

Several antifoam agents were evaluated for the ability to control foam in the production of Bacillus thuringiensis-based biopesticides using sewage sludge as a raw material. Experiments were conducted in shake flasks as well as in 15 l fermentors with controlled parameters. Polypropylene glycol (PPG), the most commonly used antifoam agent in B. thuringiensis fermentation, inhibited cell growth, sporulation and decreased the entomotoxicity yield even at a concentration of 0.1% (v/v) in sewage sludge medium. About 40% reduction in entomotoxicity was observed when PPG was used at 0.3% (v/v). The impact of PPG on sporulation and toxin synthesis in tryptic soy yeast broth (TSYB) medium was also studied. The inhibitory effects were less severe in TSYB than in sludge medium. Another silicone-based antifoam agent, “Antifoam A”, showed less severe effect on growth and stendotoxin production. The problem of the inhibitory effect of chemical antifoam agents on growth and endotoxin production was minimised substantially with the use of vegetable oils such as canola, olive, and peanut oils. Canola and peanut oil stimulated both sporulation and δ-endotoxin synthesis. The stimulus effect varies with the monounsaturated fat contents of oils. Journal of Industrial Microbiology & Biotechnology (2000) 25, 86–92. Received 09 February 2000/ Accepted in revised form 06 June 2000