Toward a consensus view on the infectious risks associated with land application of sewage sludge.

The science linking processed sewage sludge (biosolids) land application with human health has improved in the last ten years. The goal of this review is to develop a consensus view on the human health impacts associated with land-applying biosolids. Pre-existing risk studies are integrated with recent advances in biosolids pathogen exposure science and risk analysis. Other than accidental direct ingestion, the highest public risks of infection from land application are associated with airborne exposure. Multiple, independent risk assessments for enteroviruses similarly estimate the yearly probabilities of infection near 10(-4). However, the inclusion of other emerging pathogens, specifically norovirus, increases this yearly infectious risk by over 2 orders of magnitude. Quantitative microbial risk assessment for biosolids exposure more effectively operates as a tool for analyzing how exposure can be reduced rather than being used to assess "safety". Such analysis demonstrates that the tradition of monitoring pathogen quality by Salmonella spp. and enterovirus content underestimates the infectious risk to the public, and that a rigorous biosolids pathogen treatment process, rather than extending community separation distances, is the most efficient method for reducing pathogen exposure and infectious risk.

Emission rates and characterization of aerosols produced during the spreading of dewatered class B biosolids.

This study measured aerosol emission rates produced during the spreading of dewatered class B biosolids onto agricultural land. Rates were determined in multiple independent experimental runs by characterizing both the source aerosol plume geometry and aerosol concentrations of PM10, total bacteria, heterotrophic plate count bacteria (HPC), two types of biosolids indicator bacteria, endotoxin, and airborne biosolids regulated metals. These components were also measured in the bulk biosolids to allow for correlating bulk biosolids concentrations with aerosol emission rates and to produce reconstructed aerosol concentrations. The average emission rates and associated standard deviation for biosolids PM10, total bacteria, HPC, total coliforms, sulfite-reducing Clostridia, endotoxin, and total biosolids regulated metals were 10.1 +/- 8.0 (mg/s), 1.98 +/- 1.41 x 10(9) (no./s), 9.0 +/- 11.2 x 10(7) (CFU/s), 4.9 +/- 2.2 x 10(3) (CFU/ s), 6.8 +/- 3.8 x 10(3) (CFU/s), 2.1 +/- 1.8 x 10(4) (EU/s), and 36.9 +/- 31.8 (microg/s) respectively. Based on the land application rates of spreaders used in this study, an estimated 7.6 +/- 6.3 mg of biosolids were aerosolized for every 1 kg (dry weight) applied to land. Scanning electron microscopy particle size distribution analysis of the aerosols revealed that greater than 99% of the emitted particles were less than 10 microm and particle size distributions had geometric mean diameters and standard deviations near 1.1 +/- 0.97 microm. The demonstrated correlations of bulk biosolids concentrations with aerosol emission rates, and the reconstruction of aerosol concentration based on PM10 and bulk biosolids concentration provide a more fundamental, bulk biosolids-based approach for extending biosolids aerosol exposure assessment to different land application scenarios and a broader range of toxins and pathogens.

Source tracking aerosols released from land-applied class B biosolids during high-wind events.

DNA-based microbial source tracking (MST) methods were developed and used to specifically and sensitively track the unintended aerosolization of land-applied, anaerobically digested sewage sludge (biosolids) during high-wind events. Culture and phylogenetic analyses of bulk biosolids provided a basis for the development of three different MST methods. They included (i) culture- and 16S rRNA gene-based identification of Clostridium bifermentans, (ii) direct PCR amplification and sequencing of the 16S rRNA gene for an uncultured bacterium of the class Chloroflexi that is commonly present in anaerobically digested biosolids, and (iii) direct PCR amplification of a 16S rRNA gene of the phylum Euryarchaeota coupled with terminal restriction fragment length polymorphism to distinguish terminal fragments that are unique to biosolid-specific microorganisms. Each method was first validated with a broad group of bulk biosolids and soil samples to confirm the target's exclusive presence in biosolids and absence in soils. Positive responses were observed in 100% of bulk biosolid samples and in less than 11% of the bulk soils tested. Next, a sampling campaign was conducted in which all three methods were applied to aerosol samples taken upwind and downwind of fields that had recently been land applied with biosolids. When average wind speeds were greater than 5 m/s, source tracking results confirmed the presence of biosolids in 56% of the downwind samples versus 3% of the upwind samples. During these high-wind events, the biosolid concentration in downwind aerosols was between 0.1 and 2 microg/m3. The application of DNA-based source tracking to aerosol samples has confirmed that wind is a possible mechanism for the aerosolization and off-site transport of land-applied biosolids.

Source bioaerosol concentration and rRNA gene-based identification of microorganisms aerosolized at a flood irrigation wastewater reuse site.

Reuse of partially treated domestic wastewater for agricultural irrigation is a growing practice in arid regions throughout the world. A field sampling campaign to determine bioaerosol concentration, culturability, and identity at various wind speeds was conducted at a flooded wastewater irrigation site in Mexicali, Baja California, Mexico. Direct fluorescent microscopy measurements for total microorganisms, culture-based assays for heterotrophs and gram-negative enteric bacteria, and small-subunit rRNA gene-based cloning were used for microbial characterizations of aerosols and effluent wastewater samples. Bioaerosol results were divided into two wind speed regimens: (i) below 1.9 m/s, average speed 0.5 m/s, and (ii) above 1.9 m/s, average speed 4.5 m/s. Average air-borne concentration of total microorganisms, culturable heterotrophs, and gram-negative enteric bacteria were, respectively, 1.1, 4.2, and 6.2 orders of magnitude greater during the high-wind-speed regimen. Small-subunit rRNA gene clone libraries processed from samples from air and the irrigation effluent wastewater during a high-wind sampling event indicate that the majority of air clone sequences were more than 98% similar to clone sequences retrieved from the effluent wastewater sample. Overall results indicate that wind is a potential aerosolization mechanism of viable wastewater microorganisms at flood irrigation sites.