Thermophilic-anaerobic digestion to produce class A biosolids: initial full-scale studies at Hyperion Treatment Plant.

The highest quality of biosolids is called exceptional quality. To qualify for this classification, biosolids must comply with three criteria: (1) metal concentrations, (2) vector-attraction reduction, and (3) the Class A pathogen-density requirements. The City of Los Angeles Bureau of Sanitation Hyperion Treatment Plant (HTP) (Playa del Rey, California) meets the first two requirements. Thus, the objective of this study was to ensure that HTP's biosolids production would meet the Class A pathogen-reduction requirements following the time-temperature regimen for batch processing (U.S. EPA, 1993; Subsection 32, Alternative 1). Because regulations require the pathogen limits to be met at the last point of plant control, biosolids sampling was not limited to immediately after the digesters, i.e., the digester outflows. The sampling extended to several locations in HTP's postdigestion train, in particular, the last points of plant control, i.e., the truck loading facility and the farm for land application. A two-stage, thermophilic-continuous-batch process, consisting of a battery of six egg-shaped digesters, was established in late 2001 for phase I of this study and modified in early 2002 for phase II. As the biosolids were discharged from the second-stage digesters, the Salmonella sp. (pathogen) and fecal-coliform (indicator) densities were well below the limits for Class A biosolids, even though the second-stage-digester temperatures were a few degrees below the temperature required by Alternative 1. Salmonella sp. densities remained below the Class A limit at all postdigestion sampling locations. Fecal-coliform densities were also below the Class A limit at postdigestion-sampling locations, except the truck-loading facility (phases I and II) and the farm for final use of the biosolids (phase II). Although federal regulations require one of the limits for either fecal coliforms or Salmonella sp. to be met, local regulations in Kern County, California, where the biosolids are land-applied, require compliance with both bacterial limits. Additional work identified dewatering, cooling of biosolids after the dewatering centrifuges, and contamination as possible factors in the rise in density of fecal coliforms. These results provided the basis for the full conversion of HTP to the Los Angeles continuous-batch, thermophilic-anaerobic-digestion process. During later phases of testing, this process was demonstrated to produce fully disinfected biosolids at the farm for land application.     

Recurrence of fecal coliforms and Salmonella species in biosolids following thermophilic anaerobic digestion.

The U.S. Environmental Protection Agency (U.S. EPA) Part 503 Biosolids Rule requires the fecal coliform (indicator) or Salmonella species (pathogen) density requirements for Class A biosolids to be met at the last point of plant control (truck-loading facility and/or farm for land application). The three Southern Californian wastewater treatment plants in this study produced biosolids by thermophilic anaerobic digestion and all met the Class A limits for both fecal coliforms and Salmonella sp. in the digester outflow biosolids. At two plants, however, a recurrence of fecal coliforms was observed in postdigestion biosolids, which caused exceedance of the Class A limit for fecal coliforms at the truck-loading facility and farm for land application. Comparison of observations at the three plants and further laboratory tests indicated that the recurrence of fecal coliforms can possibly be related to the following combination of factors: (1) incomplete destruction of fecal coliforms during thermophilic anaerobic digestion, (2) contamination of Class A biosolids with fecal coliforms from external sources during postdigestion, (3) a large drop of the postdigestion biosolids temperature to below the maximum for fecal coliform growth, (4) an unknown effect of biosolids dewatering in centrifuges. At Hyperion Treatment Plant (City of Los Angeles, California), fecal coliform recurrence could be prevented by the following: (1) complete conversion to thermophilic operation to exclude contamination by mesophilically digested biosolids and (2) insulation and electrical heat-tracing of postdigestion train for maintaining a high biosolids temperature in postdigestion.     

Bioleaching of zinc and copper from anaerobically digested swine manure: effect of sulfur levels and solids contents.

Anaerobically digested swine manure (ADSM) generally contains high concentrations of zinc (Zn) and copper (Cu). These metals levels exceed the land application regulations of municipal biosolids of many European countries and are on the borderline of exceptional quality in the U.S. Environmental Protection Agency (U.S. EPA) 40 CFR part 503 standards. From this perspective, a series of batch tests were conducted to evaluate the feasibility of bioleaching of Zn and Cu from ADSM so that the produced biosolids could safely be applied to land. The effect of different substrate levels (sulfur) and total solids content (as high as 9%) on metal solubilization was investigated. The results showed that metal solubilization efficiency for both Zn and Cu declined significantly with an increase in total solids from 3 to 6% and then to 9% at the same substrate level. Metal solubilization increased proportionately with increases in substrate concentration up to 6% of total solids content. However, at 9% total solids content, metal solubilization was insignificant at all substrate levels tested. At a 6% total solids level and 10 000 mg/L of substrate, 94% of Zn and 72% of Cu were solubilized. Bioleaching was also found to be effective in destruction of pathogens with approximately 2.5 log-scale reduction. The residual biosolid was found to meet the Class A biosolids standands of U.S. EPA 40 CFR part 503.     

False positive fecal coliform in biosolid samples assayed using A-1 medium.

Two most probable number (MPN) methods-lauryl tryptose broth with Escherichia coli broth confirmation and direct A-1 broth incubation (A-1)--were compared for the enumeration of fecal coliform in lime-treated biosolid. Fecal coliform numbers were significantly higher using the A-1 method. Analysis of positive A-1 tubes, however, indicated that a high percentage of these were false positives. Therefore, the use of A-1 broth for 40 CFR Part 503 Pathogen Reduction (CFR, 1993) compliance testing is not recommended.     

Biosolids-amended soils: Part II. Chemical lability as a measure of contaminant bioaccessability.

Biosolids recycling by amending agricultural soils has increased significantly over the last few decades. The presence of contaminants in small, bioavailable quantities has generated concerns about health threats resulting from accumulation of potential toxins in the food chain. In this study, land application of biosolids was evaluated for environmental risk. Chemical lability tests for metals were used for the test soils and included analyses for water soluble, exchangeable, and metals extractable by the physiologically based extraction test. Chemical extractions detected slight increases in labile metal concentrations for many of the treated soils, particularly those receiving long-term applications of 5 years or more. Significantly higher metal concentrations were observed in the soils that had been exposed to biosolids before the U.S. Environmental Protection Agency (Washington, D.C.) 503 Rule (U.S. EPA, 2004) was implemented.     

Influence of staging, mean cell residence time, and thermophilic temperature on the thermophilic anaerobic digestion process.

As Class B biosolids land application has become less acceptable to many local jurisdictions, low-cost processes to achieve Class A standards have become more popular. Prominent among these low-cost processes is thermophilic anaerobic digestion. As a result, thermophilic anaerobic digestion is now a popular topic in wastewater treatment literature, but quantifiable methods for selecting a particular thermophilic process have not been offered. To provide for this need, an empirical model was developed from data collected in thermophilic anaerobic digestion studies conducted using East Bay Municipal Utility District's (Oakland, California) primary and waste activated sludge to feed both bench- and full-scale digesters. The model predicts at which thermophilic temperature and mean cell residence time (MCRT) one process will outperform or equal another, with respect to fecal coliform reduction. The different disinfection efficiencies in the different thermophilic processes might be explained by the presence or absence of high volatile acid and/or un-ionized ammonia levels in the processes' digested sludges. Data from these studies also show an apparent relationship between increased thermophilic temperatures and volatile solids destruction, and between increased temperatures and specific volatile acids production, for digesters operating at a 13-day MCRT and higher, but not for digesters operating at a 2-day MCRT.     

Bioaerosols from land-applied biosolids: issues and needs.

Bioaerosols are a vehicle for the dissemination of human and animal pathogens. Because of land-filling costs and the ban on ocean dumping of municipal biosolids, land application of biosolids and animal manure is increasing all over the globe. There is no doubt that the creation, generation, and disposal of human and animal wastes increases the aerosolization potential of a wide variety of microbial pathogens and related pollutants. In an attempt to address public health issues associated with the land application of municipal biosolids, the U.S. National Research Council (Washington, D.C.) published a report on this issue in 2002. This paper focuses on the current information and technology gaps related to estimating the public health risks associated with bioaerosols during the land application of biosolids.     

Regrowth of fecal coliforms in class A biosolids.

Eight types of Class A biosolids were tested for fecal coliform (FC) reactivation and/or regrowth at 20, 35, and 50 degrees C for 21 days. Growth of FC did not occur at 20 or 50 degrees C, but it was observed in two samples incubated at 35 degrees C after a lag period of 48 hours. In undigested biosolids, final FC concentration exceeded 10(4) MPN/g, whereas in thermophilically digested biosolids, the final FC concentration remained below 10(3) MPN/g, as FC regrowth may have been affected by the presence of the anaerobic bacterial consortium responsible for the digestion process. Fecal-coliform reactivation and regrowth within treatment plant operations seem unlikely but can occur in land application of biosolids.     

Extended acid digestion for inactivation of fecal coliforms.

The objective of this research was to establish a correlation between inactivation of fecal coliforms caused by organic acids in their unionized form in batch acid digesters and semicontinuously fed acid digesters at both mesophilic (38 degrees C) and low-mesophilic (24 degrees C) temperatures. Batch acid digesters achieved a U.S. Environmental Protection Agency Class A level of fecal coliforms within 6 to 7 days of digestion at both temperatures. Semicontinuously fed, staged, acid-digestion systems achieved Class A standards on average only at mesophilic temperature at a solids retention time of 11 days. Systems operated at low-mesophilic temperatures did not achieve Class A standards.     

Laboratory evaluation of thermophilic-anaerobic digestion to produce Class A biosolids. 1. Stabilization performance of a continuous-flow reactor at low residence time.

There is increasing interest in the United States in producing biosolids from municipal wastewater treatment that meet the criteria for Class A designation established by the U.S. Environmental Protection Agency. Class A biosolids are intended to be free of pathogens and also must meet requirements for reduction of the vector-attraction potential associated with untreated sludge. High-temperature processes are considered to produce Class A biosolids if the combination of operating temperature and treatment time exceeds minimum criteria, but this option is not applicable to mixed, continuous-flow reactors. Such reactors, or any combination of reactors that does not meet the holding time requirement at a specific temperature, must be demonstrated to inactivate pathogens to levels consistent with the Class A criteria. This study was designed to evaluate pathogen inactivation by thermophilic anaerobic digestion in a mixed, continuous-flow reactor followed by batch or plug-flow treatment. In this first of a two-part series, we describe the performance of a continuous-flow laboratory reactor with respect to physical and chemical operating parameters; microbial inactivation in the combined continuous-flow and batch treatment system is described in the second part. Sludges from three different sources were treated at 53 degrees C, while sludge from one of the sources was also treated at 55 and 51 degrees C. Relatively short hydraulic retention times (four to six days) were used to represent a conservative operating condition with respect to pathogen inactivation. Treatment of a fermented primary sludge led to an average volatile-solids (VS) destruction efficiency of 45%, while VS destruction for the other two sources was near or below 38%, the Class A criterion for vector attraction reduction. Consistent with other studies on thermophilic anaerobic digestion of sludges at short residence times, effluent concentrations of volatile fatty acids (VFAs) were relatively high. Also consistent with other studies, the most abundant VFA in the effluent was propionate. Gas production ranged from 0.3 to 0.5 m3/kg VS fed and from 0.8 to 1.3 m3/kg VS destroyed.     

Development of a hazardous air pollutants monitoring program using the Data Quality Objectives process

To effectively reduce the environmental compliance costs associated with meeting hazardous air pollutant emission requirements, the U.S. Environmental Protection Agency's Data Quality Objective (DQO) process has been proposed as a suitable framework for establishing a defensible monitoring program. Through the use of a hazardous materials pilot study, the variability in the composite vapor pressure for regulated handwipe cleaning solvents was established. These results served as inputs to the DQO process, which identified that for facility decision-makers to claim with a 99% confidence level that the facility is in compliance with the National Emission Standards for Hazardous Air Pollutants (NESHAP), a minimum of 12 handwipe cleaning solvent compliance samples (taken at random every 6 months) must have a composite vapor pressure equal to or below the regulatory limit of 45 mmHg at 20 degrees C. Implementation of the DQO-based compliance-sampling plan eliminates the need for an affected facility to sample all regulated handwipe cleaning solvents while still maintaining a reasonably high level of confidence in the compliance status of its regulated sources. The approach described for designing a defensible compliance sampling plan can be extended to other aspects of the aerospace NESHAP rule, including compliance sampling for surface coating, chemical depainting, and hazardous waste disposal.     

Chemical speciation and extractability of Zn,Cu and Cd in two contrasting biosolids-amended clay soils

An incubation experiment was conducted to study the chemical speciation and extractability of three heavy metals in two contrasting biosolids-amended clay soils. One was a paddy soil of pH 7.8 and the other was a red soil of pH 4.7 collected from a fallow field. Anaerobically digested biosolids were mixed with each of the two soils at three rates: 20, 40 and 60 g kg(-1) soil (DM basis), and unamended controls were also prepared. The biosolids-amended and control soils were incubated at 70% of water holding capacity at 25 degrees C for 50 days. Separate subsamples were extracted with three single extractants and a three-step sequential extraction procedure representing acetic acid (HOAc)-soluble, reducible and oxidisable fractions to investigate the extractability and speciation of the heavy metals. As would be expected, there were good relationships between biosolids application rate and metal concentrations in the biosolids-amended soils. The three heavy metals had different extractabilities and chemical speciation in the two biosolids-amended soils. Ethylene diamine tetraacetic acid extracted more Cu, Zn and Cd than did the other two single extractants. The oxidisable fraction was the major fraction for Cu in both biosolids-amended soils and the HOAc-soluble and reducible fractions accounted for most of the Zn. In contrast, Cd was present mainly in the reducible fraction. The results are discussed in relation to the mobility and bioavailability of the metals in polluted soils.     

Maintaining high-volume, low-pressure surface-coating regulatory compliance using the U.s. Environmental Protection Agency's data quality objectives process

To effectively reduce the environmental compliance costs associated with meeting specific requirements under the Aerospace Manufacturing and Rework Facility's National Emission Standard for Hazardous Air Pollutants rule, the U.S. Environmental Protection Agency's (EPA) Data Quality Objective (DQO) process has been proposed as a suitable framework for developing a scientifically defensible surface compliance monitoring program. By estimating the variability associated with the air cap pressure of high- volume, low-pressure (HVLP) surface-coating spray equipment, the number of monitoring samples necessary for an affected facility to claim compliance with a desired statistical confidence level was established. Using data taken from the pilot test facility, the DQO process indicated that the mean of at least 21 HVLP air cap pressure samples taken over the compliance period must be < or = 10 pounds per square inch (psig) gauge for the facility to claim regulatory compliance with 99.99% statistical confidence. Fewer compliance samples could be taken, but that decision would lead to a commensurate reduction in the compliance confidence level. Implementation of the DQO-based compliance sampling plan eliminates the need for an affected facility to sample all regulated HVLP surface-coating processes while still maintaining a high level of compliance assurance.     

Maintaining High-Volume,Low-Pressure Surface-Coating Regulatory Compliance Using the U.S. Environmental Protection Agency’s Data Quality Objectives Process

Abstract

To effectively reduce the environmental compliance costs associated with meeting specific requirements under the Aerospace Manufacturing and Rework Facility’s National Emission Standard for Hazardous Air Pollutants rule, the U.S. Environmental Protection Agency’s (EPA) Data Quality Objective (DQO) process has been proposed as a suitable framework for developing a scientifically defensible surface compliance monitoring program. By estimating the variability associated with the air cap pressure of high-volume, low-pressure (HVLP) surface-coating spray equipment, the number of monitoring samples necessary for an affected facility to claim compliance with a desired statistical confidence level was established. Using data taken from the pilot test facility, the DQO process indicated that the mean of at least 21 HVLP air cap pressure samples taken over the compliance period must be ≤10 pounds per square inch (psig) gauge for the facility to claim regulatory compliance with 99.99% statistical confidence. Fewer compliance samples could be taken, but that decision would lead to a commensurate reduction in the compliance confidence level. Implementation of the DQO-based compliance sampling plan eliminates the need for an affected facility to sample all regulated HVLP surface-coating processes while still maintaining a high level of compliance assurance.     

Technological options for the management of biosolids

BACKGROUND, AIM, AND SCOPE: Large quantities of biosolids (sewage sludge), which are produced from municipal wastewater treatment, are ever-increasing because of the commissioning of new treatment plants and continuous upgrades of the existing facilities. A large proportion of biosolids are currently landfilled. With increasing pressure from regulators and the general public, landfilling of biosolids is being phased out in many countries because of potential secondary pollution caused by leachate and the emission of methane, a potent greenhouse gas. Biosolids contain nutrients and energy that can be used beneficially. Significant efforts have been made recently to develop new technologies to manage biosolids and make useful products from them. In this paper, we provide a review of the technologies in biosolids management. MATERIALS AND METHODS: A survey of literature was conducted. RESULTS: At present, the most common beneficial use of biosolids is agricultural land application because of inherent fertilizer values found in biosolids. Expansion of land application, however, may be limited in the future because of more stringent regulatory requirements and public concern about food chain contamination in some countries. Perceived as a green energy source, the combustion of biosolids has received renewed interest. Anaerobic digestion is generally a more effective method than incineration for energy recovery, and digested biosolids are suitable for further beneficial use through land application. Although conventional incineration systems for biosolid management generally consume more energy than they produce because of the high moisture content in the biosolids, it is expected that more combustion systems, either monocombustion or cocombustion, will be built to cope with the increasing quantity of biosolids. DISCUSSION: Under the increasingly popular low-carbon economy policy, biosolids may be recognized as a renewable fuel and be eligible for 'carbon credits'. Because ash can be used to manufacture construction materials, combustion can provide a complete management for biosolids. A number of advanced thermal conversion technologies (e.g., supercritical water oxidation process and pyrolysis) are under development for biosolids management with a goal to generate useful products, such as higher quality fuels and recovery of phosphorus. With an ever-increasing demand for renewable energy, growing bioenergy crops and forests using biosolids as a fertilizer and soil amendment can not only contribute to the low-carbon economy but also maximize the nutrient and carbon value of the biosolids. CONCLUSIONS: Land application of biosolids achieves a complete reuse of its nutrients and organic carbon at a relatively low cost. Therefore, land application should become a preferred management option where there is available land, the quality of biosolids meet regulatory requirements, and it is socially acceptable. Intensive energy cropping and forest production using biosolids can help us meet the ever-increasing demand for renewable energy, which can eliminate the contamination potential for food sources, a common social concern about land application of biosolids. In recent years, increasing numbers of national and local governments have adopted more stringent regulations toward biosolid management. Under such a political climate, biosolids producers will have to develop multireuse strategies for biosolids to avoid being caught because a single route management practice might be under pressure at a short notice. Conventional incineration systems for biosolids management generally consume more energy than they produce and, although by-products may be used in manufacturing, this process cannot be regarded as a beneficial use of biosolids. However, biosolids are likely to become a source of renewable energy and produce 'carbon credits' under the increasingly popular, low-carbon economy policy. RECOMMENDATIONS AND PERSPECTIVES: To manage biosolids in a sustainable manner, there is a need for further research in the following areas: achieving a higher degree of public understanding and acceptance for the beneficial use of biosolids, developing cost-efficient and effective thermal conversions for direct energy recovery from biosolids, advancing technology for phosphorus recovery, and selecting or breeding crops for efficient biofuel production.     

High-solids anaerobic digestion of municipal sludge pretreated by thermal hydrolysis

High-solids anaerobic digestion can consistently achieve 55 to 60% volatile solids destruction after thermal hydrolysis pretreatment, which reduces its viscosity and increases the fraction of soluble organic matter. For feed sludge with total solids concentrations between 6.8 and 8.2%, the process is stable at hydraulic retention times of 9 to 12 days, significantly increasing the treatment capacity of existing digesters or, in treatment plants without spare capacity, helping to postpone, reduce, or even avoid costly infrastructure investments. Process stability is related to the high concentration of soluble organic matter in the digesters. High-solids temperature-phased digestion appears to be superior to high-solids mesophilic digestion, with respect to process flexibility and stability, biosolids stabilization, and biogas generation, although ammonia inhibition may have occurred. Implementation of high-solids digestion could significantly reduce operation and maintenance costs of solids-handling operations.     

Biosolids-amended soils: Part I. Effect of biosolids application on soil quality and ecotoxicity.

Investigations of potential risk from biosolids generally indicate that land application does not threaten human or ecosystem health, but questions continue to arise concerning the environmental effects of this practice. This research project was initiated to evaluate ecotoxicity resulting from the amendment of soils with biosolids from municipal wastewater treatment plants. Toxicity was evaluated using standard tests, including earthworm mortality, growth, and reproduction; seedling germination and root elongation; microbial respiration; and nematode mortality and reproduction. Nineteen municipal wastewater treatment plants were identified to participate in an initial screening of toxicity, and five were chosen for a more detailed evaluation. In addition, two soils with historically high applications of high-metal biosolids were evaluated. Contaminants examined were zinc, copper, nickel, chromium, arsenic, cadmium, lead, and coplanar polychlorinated biphenyls (PCBs). Single applications had no effect on soil metal concentrations. Coplanar PCBs were not detectable in any of the soils or biosolids. All target organisms were sensitive to reference toxicants. Limited toxicity was observed in a small number of the amended soils, but no patterns emerged. Approximately one-half of the negative effects of biosolids on bioindicators could be attributed to routine properties, such as slight depression of pH and/or elevated salinity. None of the accumulated metal concentrations were excessive, and most would not be considered elevated. These observations suggest that current regulations for application of biosolids to soils are providing adequate ecosystem protection.     

Anaerobic codigestion of municipal, farm, and industrial organic wastes: a survey of recent literature.

Codigestion of organic wastes is a technology that is increasingly being applied for simultaneous treatment of several solid and liquid organic wastes. The main advantages of this technology are improved methane yield because of the supply of additional nutrients from the codigestates and more efficient use of equipment and cost-sharing by processing multiple waste streams in a single facility. Many municipal wastewater treatment plants (WWTPs) in industrialized countries currently process wastewater sludge in large digesters. Codigestion of organic wastes with municipal wastewater sludge can increase digester gas production and provide savings in the overall energy costs of plant operations. Methane recovery also helps to reduce the emission of greenhouse gases to the atmosphere. The goal of this literature survey was to summarize the research conducted in the last four years on anaerobic codigestion to identify applications of codigestion at WWTPs. Because the solids content in municipal wastewater sludge is low, this survey only focuses on codigestion processes operated at relative low solids content (slurry mode). Semi-solid or solid codigestion processes were not included. Municipal wastewater sludge, the organic fraction of municipal solid waste, and cattle manure (CAM) are the main wastes most often used in codigestion processes. Wastes that are codigested with these main wastes are wood wastes, industrial organic wastes, and farm wastes. These are referred to in this survey as codigestates. The literature provides many laboratory studies (batch assays and bench-scale digesters) that assess the digestibility of codigestates and evaluate the performance and monitoring of codigestion, inhibition of digestion by codigestates, the design of the process (e.g., single-stage or two-stage processes), and the operation temperature (e.g., mesophilic or thermophilic). Only a few reports on pilot- and full-scale studies were found. These evaluate general process performance and pretreatment of codigestates, energy production, and treatment costs.     

Comparative study of hotplate wet digestion methods for the determination of mercury in biosolids

The re-use of biosolids is becoming increasingly popular for land applications. However, biosolids may contain elevated levels of metals and metalloids (including mercury) relative to background environmental concentrations. Consequently, reliable mercury analysis is important to allow classification of biosolids and to determine appropriate options for beneficial uses. This paper reports on a comparative study of 12 hotplate wet digestion methods for their suitability for the determination of mercury in biosolids. The methods were applied to mercury biosolids samples from four localities of two different sewage treatment plants in the State of Victoria, Australia. Samples were also spiked with methylmercury chloride and mercury sulphide to evaluate the Hg recovery in each hotplate digestion method. Aqua regia (HCl:HNO(3)=3:1), reverse aqua regia (HCl:HNO(3)=1:3), nitric, hydrochloric, sulphuric acid and their combinations with or without hydrogen peroxide were studied as wet digestion solutions. The method providing the best mercury recoveries was optimized. Under optimal conditions the corresponding analytical procedure consisted of 1h pre-digestion of 0.4 g biosolids sample with 10 ml reverse aqua regia with temperature increasing to 110 degrees C and 3h digestion at this temperature. In the last 10 min of the digestion step, 2 ml hydrogen peroxide were added to ensure complete decomposition of all mercury containing compounds. After filtering and dilution with deionised water (1:10), the concentration of mercury was determined by cold vapour atomic absorption spectrometry. It is expected, that the wet acid digestion method developed in this study will be also applicable to biosolids from other sewage treatment plants and to other types of solid mercury samples with elevated levels of organic matter.     

The relationship between mixed-liquor particle size and solids retention time in the activated sludge process

Particle size distribution (PSD) analysis was used to evaluate the quality of mixed liquors collected from different activated sludge process modifications (i.e., conventional activated sludge, modified Ludzack-Ettinger, high-purity oxygen, step-anoxic, and oxidation ditch). An experiment protocol was developed to define the allowable sample holding time and provide representative and repeatable results. Samples of 26 treatment plants, with a total of 37 samples, were tested. A new indicator, called mean particle size (MPS), was introduced to describe the integrated mean particle size. The results of MPSs of three cut-off sizes (0.5 to 50, 100, and 200 microm) showed that the average size of mixed-liquor biosolids increased with increasing solids retention time (SRT), and the number of particles in the sedimentation supernatant decreased with increasing SRT. Particle deflocculation occurred after excessive sample holding time, and analysis within 12 hours generally eliminated sample holding problems. The results provide a methodology using PSD for characterizing mixed-liquor biosolids.