John P. Brooks

A national study on the residential impact of biological aerosols from the land application of biosolids

Aims:  The purpose of this study was to evaluate the community risk of infection from bioaerosols to residents living near biosolids land application sites.

Estimation of bioaerosol risk of infection to residents adjacent to a land applied biosolids site using an empirically derived transport model

Aim:  The purpose of this study was to develop an empirically derived transport model, which could be used to predict downwind concentrations of viruses and bacteria during land application of liquid biosolids and subsequently assess microbial risk associated with this practice.

Pathogens in Biosolids

The world population of 6.8 billion people all produce sewage. In the developed world most of this is treated by the activated sludge process, which results in large volumes of sludge or biosolids being produced (NRC, 2002). This results in millions of tons of biosolids produced each year in the United States, which must either be disposed of or recycled in some manner. Land application has been seen as the most economical and beneficial way of handling biosolids. Biosolids that result from municipal wastewater treatment processes contain organic matter and nutrients that, when properly treated and applied to farmland, can improve the productivity of soils or enhance revegetation of disturbed ecosystems. However, besides the documented benefits of land application, there are also potential hazards, which have caused the public response to the practice to be mixed. Here we review one of the potential hazards associated with biosolids and its land application, namely human pathogens associated with biosolids.

Rainfall simulation in greenhouse microcosms to assess bacterial-associated runoff from land-applied poultry litter.

Runoff water following a rain event is one possible source of environmental contamination after a manure application. This greenhouse study used a rainfall simulator to determine bacterial-associated runoff from troughs of common bermudagrass [Cynodon dactylon (L.) Pers.] that were treated with P-based, N-based, and N plus lime rates of poultry (Gallus gallus) litter, recommended inorganic fertilizer, and control. Total heterotrophic plate count (HPC) bacteria, total and thermotolerant coliforms, enterococci, staphylococci, Clostridium perfringens, Salmonella, and Campylobacter, as well as antibiotic resistance profiles for the staphylococci and enterococci isolates were all monitored in runoff waters. Analysis following five rainfall events indicated that staphylococci, enterococci, and clostridia levels were related to manure application rate. Runoff release of staphylococci, enterococci, and C. perfringens were approximately 3 to 6 log10 greater in litter vs. control treatment. In addition, traditional indicators such as thermotolerant and total coliforms performed poorly as fecal indicators. Some isolated enterococci demonstrated increased antibiotic resistance to polymixin b and/or select aminoglyocosides, while many staphylococci were susceptible to most antimicrobials tested. Results indicated poultry litter application can lead to microbial runoff following simulated rain events. Future studies should focus on the use of staphylococci, enterococci, and C. perfringens as indicators.

Microbial and antibiotic resistant constituents associated with biological aerosols and poultry litter within a commercial poultry house

Poultry are known to harbor antibiotic resistant and pathogenic bacteria, and as such poultry litter and poultry house air can be contaminated with these bacteria. However, the presence of antibiotic resistant bacteria in biological aerosols and litter is largely not understood. The purpose of this study was to determine the amount of aerosolized bacteria and endotoxin, particularly fecal indicators, staphylococci, and enterococci, associated with poultry house and outdoor air. Aerosol samples were collected at multiple locations on the farm and in the house. Antibiotic resistance was investigated using the Kirby Bauer method on selected isolates using twelve different antibiotics spanning both narrow to broad spectrums of effectiveness. Overall there was a cyclical increase in bacterial concentrations as flocks progressed from pre-flock to late-flock, with >2 orders magnitude lower concentration during pre-flock periods (no chickens), in both the litter and aerosol samples. The house environment provided for significantly concentrated bacterial and endotoxin levels. It was estimated that Staphylococcus bacteria accounted for at least 90% of cultured aerobic bacteria and culture-independent 16S rRNA analyses demonstrated that significant population changes occurred from pre- to late-flock. Rarely was an isolate resistant to more than 4 antibiotic classes; however there was a trend upwards in overall resistance of enterococci as the flock cycle progressed. It appears that although levels of antibiotic resistant bacteria were highly concentrated within the house, levels were much lower outside of the house, and very little house escape occurred.

Land application of manure and Class B biosolids: an occupational and public quantitative microbial risk assessment.

Land application is a practical use of municipal Class B biosolids and manure that also promotes soil fertility and productivity. To date, no study exists comparing biosolids to manure microbial risks. This study used quantitative microbial risk assessment to estimate pathogen risks from occupational and public exposures during scenarios involving fomite, soil, crop, and aerosol exposures. Greatest one-time risks were from direct consumption of contaminated soil or exposure to fomites, with one-time risks greater than 10. Recent contamination and high exposures doses increased most risks. and enteric viruses provided the greatest single risks for most scenarios, particularly in the short term. All pathogen risks were decreased with time, 1 d to14 mo between land application and exposure; decreases in risk were typically over six orders of magnitude beyond 30 d. Nearly all risks were reduced to below 10 when using a 4-mo harvest delay for crop consumption. Occupational, more direct risks were greater than indirect public risks, which often occur after time and dilution have reduced pathogen loads to tolerable levels. Comparison of risks by pathogen group confirmed greater bacterial risks from manure, whereas viral risks were exclusive to biosolids. A direct comparison of the two residual types showed that biosolids use had greater risk because of the high infectivity of viruses, whereas the presence of environmentally recalcitrant pathogens such as and maintained manure risk. Direct comparisons of shared pathogens resulted in greater manure risks. Overall, it appears that in the short term, risks were high for both types of residuals, but given treatment, attenuation, and dilution, risks can be reduced to near-insignificant levels. That being said, limited data sets, dose exposures, site-specific inactivation rates, pathogen spikes, environmental change, regrowth, and wildlife will increase risk and uncertainty and remain areas poorly understood.

Estimated Occupational Risk from Bioaerosols Generated during Land Application of Class B Biosolids

Some speculate that bioaerosols from land application of biosolids pose occupational risks, but few studies have assessed aerosolization of microorganisms from biosolids or estimated occupational risks of infection. This study investigated levels of microorganisms in air immediately downwind of land application operations and estimated occupational risks from aerosolized microorganisms. In all, more than 300 air samples were collected downwind of biosolids application sites at various locations within the United States. Coliform bacteria, coliphages, and heterotrophic plate count (HPC) bacteria were enumerated from air and biosolids at each site. Concentrations of coliforms relative to Salmonella and concentrations of coliphage relative to enteroviruses in biosolids were used, in conjunction with levels of coliforms and coliphages measured in air during this study, to estimate exposure to Salmonella and enteroviruses in air. The HPC bacteria were ubiquitous in air near land application sites whether or not biosolids were being applied, and concentrations were positively correlated to windspeed. Coliform bacteria were detected only when biosolids were being applied to land or loaded into land applicators. Coliphages were detected in few air samples, and only when biosolids were being loaded into land applicators. In general, environmental parameters had little impact on concentrations of microorganisms in air immediately downwind of land application. The method of land application was most correlated to aerosolization. From this large body of data, the occupational risk of infection from bioaerosols was estimated to be 0.78 to 2.1%/yr. Extraordinary exposure scenarios carried an estimated annual risk of infection of up to 34%, with viruses posing the greatest threat. Risks from aerosolized microorganisms at biosolids land application sites appear to be lower than those at wastewater treatment plants, based on previously reported literature.

Long-term effects of land application of Class B biosolids on the soil microbial populations, pathogens, and activity.

This study evaluated the influence of 20 annual land applications of Class B biosolids on the soil microbial community. The potential benefits and hazards of land application were evaluated by analysis of surface soil samples collected following the 20th land application of biosolids. The study was initiated in 1986 at the University of Arizona Marana Agricultural Center, 21 miles north of Tucson, AZ. The final application of biosolids was in March 2005, followed by growth of cotton (Gossypium hirsutum L.) from April through November 2005. Surface soil samples (0-30 cm) were collected monthly from March 2005, 2 wk after the final biosolids application, through December 2005, and analyzed for soil microbial numbers. December samples were analyzed for additional soil microbial properties. Data show that land application of Class B biosolids had no significant long-term effect on indigenous soil microbial numbers including bacteria, actinomycetes, and fungi compared to unamended control plots. Importantly, no bacterial or viral pathogens were detected in soil samples collected from biosolid amended plots in December (10 mo after the last land application) demonstrating that pathogens introduced via Class B biosolids only survived in soil transiently. However, plots that received biosolids had significantly higher microbial activity or potential for microbial transformations, including nitrification, sulfur oxidation, and dehydrogenase activity, than control plots and plots receiving inorganic fertilizers. Overall, the 20 annual land applications showed no long-term adverse effects, and therefore, this study documents that land application of biosolids at this particular site was sustainable throughout the 20-yr period, with respect to soil microbial properties.

Characterization of Selected Nutrients and Bacteria from Anaerobic Swine Manure Lagoons on Sow, Nursery, and Finisher Farms in the Mid-South USA

Swine (Sus scrofa domestica) production in the Mid-South USA comprises sow, nursery, and finisher farms. A 2007 packing plant closure started a regional shift from finisher to sow and nursery farms. Changes in manure stored in lagoons and land-applied as fertilizer were expected but were unknown because nutrient and bacterial levels had not been characterized by farm type. The objectives of this study were to quantify selected nutrients and bacteria, compare levels by farm types, and project impacts of production shifts. Nutrients and bacteria were characterized in 17 sow, 10 nursery, and 10 finisher farm lagoons. Total and thermotolerant coliforms, Escherichia coli (Migula) Castellani and Chalmers, Enterococcus spp., Clostridium perfringens (Veillon and Zuber) Hauduroy et al., Campylobacter spp., Listeria spp., and Salmonella spp. were evaluated. Highest levels were from total coliforms (1.4- 5.7x10(5) cfu 100 mL(-1)), which occurred with E. coli, Campylobacter spp., C. perfringens, and Enterococcus spp., in every lagoon and virtually every sample. Lowest levels were from Listeria spp. and Salmonella spp. (<or=1.3x10(2) most probable number [MPN] 100 mL(-1)), detected in 81 and 89% of lagoons and 68 and 64% of samples, respectively. Sow farm levels were higher for all except Listeria spp. and Salmonella spp., which were lower (1.4x10(1) and 2.8x10(1) MPN 100 mL(-1), respectively) and only slightly below their respective levels from nursery farms (1.1x10(2) and 3.4x10(1) MPN 100 mL(-1)). Shifting from finisher to nursery farm would not affect bacterial levels, but shifting to sows would. Either shift would reduce NPK and N:P and suggest modification of nutrient management plans.

Bacterial populations within copper mine tailings: long-term effects of amendment with Class A biosolids.

This study evaluates the effect of surface application of dried Class A biosolids on microbial populations within copper mine tailings.