Quantitative microbial risk assessment for ingestion of antibiotic resistance genes from private wells contaminated by human and livestock fecal sources.

We used quantitative microbial risk assessment to estimate ingestion risk for intI1, erm(B), sul1, tet(A), tet(W), and tet(X) in private wells contaminated by human and/or livestock feces. Genes were quantified with five human-specific and six bovine-specific microbial source-tracking (MST) markers in 138 well-water samples from a rural Wisconsin county. Daily ingestion risk (probability of swallowing ≥1 gene) was based on daily water consumption and a Poisson exposure model. Calculations were stratified by MST source and soil depth over the aquifer where wells were drilled. Relative ingestion risk was estimated using wells with no MST detections and >6.1 m soil depth as a referent category. Daily ingestion risk varied from 0 to 8.8 × 10-1 by gene and fecal source (i.e., human or bovine). The estimated number of residents ingesting target genes from private wells varied from 910 (tet(A)) to 1,500 (intI1 and tet(X)) per day out of 12,000 total. Relative risk of tet(A) ingestion was significantly higher in wells with MST markers detected, including wells with ≤6.1 m soil depth contaminated by bovine markers (2.2 [90% CI: 1.1-4.7]), wells with >6.1 m soil depth contaminated by bovine markers (1.8 [1.002-3.9]), and wells with ≤6.1 m soil depth contaminated by bovine and human markers simultaneously (3.1 [1.7-6.5]). Antibiotic resistance genes (ARGs) were not necessarily present in viable microorganisms, and ingestion is not directly associated with infection. However, results illustrate relative contributions of human and livestock fecal sources to ARG exposure and highlight rural groundwater as a significant point of exposure.IMPORTANCEAntibiotic resistance is a global public health challenge with well-known environmental dimensions, but quantitative analyses of the roles played by various natural environments in transmission of antibiotic resistance are lacking, particularly for drinking water. This study assesses risk of ingestion for several antibiotic resistance genes (ARGs) and the class 1 integron gene (intI1) in drinking water from private wells in a rural area of northeast Wisconsin, United States. Results allow comparison of drinking water as an exposure route for antibiotic resistance relative to other routes like food and recreational water. They also enable a comparison of the importance of human versus livestock fecal sources in the study area. Our study demonstrates the previously unrecognized importance of untreated rural drinking water as an exposure route for antibiotic resistance and identifies bovine fecal material as an important exposure factor in the study setting.

National Greenhouse Gas Emission Reduction Potential from Adopting Anaerobic Digestion on Large-Scale Dairy Farms in the United States.

Waste-to-energy systems can provide a functional demonstration of the economic and environmental benefits of circularity, innovation, and reimagining existing systems. This study offers a robust quantification of the greenhouse gas (GHG) emission reduction potential of the adoption of anaerobic digestion (AD) technology on applicable large-scale dairy farms in the contiguous United States. GHG reduction estimates were developed through a robust life cycle modeling framework paired with sensitivity and uncertainty analyses. Twenty dairy configurations were modeled to capture important differences in housing and manure management practices, applicable AD technologies, regional climates, storage cleanout schedules, and methods of land application. Monte Carlo results for the 90% confidence interval illustrate the potential for AD adoption to reduce GHG emissions from the large-scale dairy industry by 2.45-3.52 MMT of CO2-eq per year considering biogas use only in renewable natural gas programs and as much as 4.53-6.46 MMT of CO2-eq per year with combined heat and power as an additional biogas use case. At the farm level, AD technology may reduce GHG emissions from manure management systems by 58.1-79.8% depending on the region. Discussion focuses on regional differences in GHG emissions from manure management strategies and the challenges and opportunities surrounding AD adoption.

Determining the prevalence of Escherichia coli, Salmonella, and shiga toxin-producing Escherichia coli in manure of dairy lagoons.

AIM: The aim of this study was to determine the prevalence of microbial pathogens in manure of dairy lagoons in California. METHODS AND RESULTS: To determine pathogens in dairy manure stored in anaerobic lagoons of dairy farm, an extensive field study was conducted across California to sample manure from 20 dairy farms. Samples were analyzed to determine the prevalence of indicator Escherichia coli, Shiga toxin producing E. coli (STEC), Salmonella, and E. coli O157: H7. To test the E. coli, STEC, and Salmonella, we used agar culture-based method followed by polymerase chain reaction (PCR) method. In addition, a real- time PCR based method was used to determine the presence of E coli O157: H7. Study demonstrated that the prevalence of Salmonella in manure sample is lower than E. coli. The presence of Salmonella was found in 2.26% of the samples, and both the culture-based and PCR methods yielded comparable outcomes in detecting Salmonella. Moreover, ∼11.30% of the total samples out of the 177 were identified as positive for STEC by qPCR. CONCLUSION: These findings demonstrate that indicator E. coli are abundantly present in anaerobic lagoons. However, the presence of STEC, and Salmonella is substantially low.

Micro-positive pressure significantly decreases greenhouse gas emissions by regulating archaeal community during industrial-scale dairy manure composting.

In this study, the effects of micro-positive pressure formed by covering with a semipermeable membrane in the heating phase of dairy manure composting on greenhouse gas emissions and the mechanism of reducing methane emissions by the archaeal community were investigated. A large-scale experiment was conducted with semipermeable membrane-covered composting (SMC), forced aeration composting (FAC), and traditional static composting (TSC) groups. The results showed that the oxygen concentration and methanogen abundance were key factors in regulating methane emissions. In the heating phase of SMC, the micro-positive pressure could enhance the O2 utilization rate and heating rate, resulting in Methanobrevibacter and Methanobacterium greatly decreasing, and the abundance of mcrA decreased by 90.03%, while that of pmoA did not increase. Compared with FAC and TSC, the cumulative methane emissions in SMC decreased by 51.75% and 96.04%, respectively. Therefore, the micro-positive pressure could effectively reduce greenhouse gas emissions by inhibiting the growth of methanogens.

Partial wet oxidation of dairy manure as a pretreatment process to produce acetic acid 'a Source Growth of Methanogens'.

Wet oxidation can be an effective process for the pretreatment of complex biomass such as lignocellulose. However, studies on the use of wet oxidation for treating solid waste such as dairy manure are limited. The use of partial wet oxidation to convert dairy manure into low molecular weight carboxylic acids as final products were investigated. This work focuses on the performance of the sub-critical wet oxidation treatment of dairy cattle manure as a conversion/pretreatment process to release matter from the lignocellulosic fraction rather than a destructive process. The operating conditions were controlled at the short residence time and optimal temperature in the presence of oxygen under a pressure of 120 psi. The thermal hydrolysis under wet oxidation significantly affected conversion manure slurry into organic acids. The concentration of acetic acid reached 1778 mg L-1, achieved at 190°C (60 minutes reaction time) as the reaction temperature increased within the range of 150°C-200°C, total organic carbon was reduced and monomers in the process liquids decreased. On the other hand, soluble COD in process liquids increased with an increment in reaction temperature. The results provide insights into technical options to pretreat dairy manure to improve biochemical conversion yield.

Abiotic and Biotic Drivers of Soil Fungal Communities in Response to Dairy Manure Amendment.

Modern agriculture often relies on large inputs of synthetic fertilizers to maximize crop yield potential, yet their intensive use has led to nutrient losses and impaired soil health. Alternatively, manure amendments provide plant available nutrients, build organic carbon, and enhance soil health. However, we lack a clear understanding of how consistently manure impacts fungal communities, the mechanisms via which manure impacts soil fungi, and the fate of manure-borne fungi in soils. We assembled soil microcosms using five soils to investigate how manure amendments impact fungal communities over a 60-day incubation. Further, we used autoclaving treatments of soils and manure to determine if observed changes in soil fungal communities were due to abiotic or biotic properties, and if indigenous soil communities constrained colonization of manure-borne fungi. We found that manure amended soil fungal communities diverged from nonamended communities over time, often in concert with a reduction in diversity. Fungal communities responded to live and autoclaved manure in a similar manner, suggesting that abiotic forces are primarily responsible for the observed dynamics. Finally, manure-borne fungi declined quickly in both live and autoclaved soil, indicating that the soil environment is unsuitable for their survival. IMPORTANCE Manure amendments in agricultural systems can impact soil microbial communities via supplying growth substrates for indigenous microbes or by introducing manure-borne taxa. This study explores the consistency of these impacts on soil fungal communities and the relative importance of abiotic and biotic drivers across distinct soils. Different fungal taxa responded to manure among distinct soils, and shifts in soil fungal communities were driven largely by abiotic factors, rather than introduced microbes. This work demonstrates that manure may have inconsistent impacts on indigenous soil fungi, and that abiotic properties of soils render them largely resistant to invasion by manure-borne fungi.

Effects of different composting methods on antibiotic-resistant bacteria, antibiotic resistance genes, and microbial diversity in dairy cattle manures.

Composting is a common practice used for treating animal manures before they are used as organic fertilizers for crop production. Whether composting can effectively reduce microbial pathogens and antibiotic resistance genes remain poorly understood. In this study, we compared 3 different dairy manure composting methods-anaerobic fermentation (AF), static compost (SC), and organic fertilizer production (OFP)-for their effects on antibiotic-resistant bacteria, antibiotic resistance genes, and microbial community diversity in the treated manures. The 3 composting methods produced variable and distinct effects on antibiotic-resistant bacteria, zoonotic bacteria, and resistance genes, some of which were decreased and others of which showed no significant changes during composting. Particularly, SC and OFP reduced chloramphenicol resistance gene fexA and opportunistic pathogen Vibrio fluvialis, whereas AF significantly reduced tetracycline resistance gene tetB and opportunistic pathogens Enterococcus faecium and Escherichia fergusonii. The compositions of microbial communities varied significantly during the composting processes, and there were significant differences between the 3 composting methods. In all 3 composts, the dominant phyla were Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria. Interestingly, Firmicutes, Proteobacteria, and Bacteroidetes remained stable in the entire AF process, whereas they were dominated at the beginning, decreased at the early stage of composting, and rebounded at the later stage during SC and OFP. In general, SC and OFP produced a more profound effect than AF on microbial community diversities, pathogens, and dominant species. Additionally, Enterococcus aquimarinus was isolated from AF for the first time. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States function prediction analysis indicated that the genes related to membrane transport and amino acid metabolism were abundant in the 3 composts. The metabolism of amino acids, lipids, and carbohydrates increased as composting progressed. The biosynthesis of antibiotics was enhanced after fermentation in the 3 composting methods, and the increase in the SC was the most obvious. These results reveal dynamic changes in antibiotic-resistant bacteria, antibiotic resistance genes, microbial community composition, and function succession in different dairy manure composts and provide useful information for further optimization of composting practices.

Compaction effects on greenhouse gas and ammonia emissions from solid dairy manure.

Waste management practices of solid dairy manures were evaluated under controlled conditions to study gas transport and emission inside manure piles. Three applied stresses and three moisture contents were tested to represent manure conditions managed at various pile depths. A Fourier-transform infrared spectroscopy monitor measured concentrations of greenhouses gases [methane, carbon dioxide, and nitrous oxide] and ammonia as part of gas flux rate calculations. Results showed that carbon dioxide dominated the greenhouse gas emissions under all test conditions. Gas transfer, primarily diffusion, was facilitated by manure with high mechanical strength and high permeability. Gas emission rates reduced dramatically when moisture content increased in manure with high water holding capacity, while compaction treatments did not as strongly affect the gas emission rates. Results provide fundamental insights into management strategies for reducing gas emissions from solid dairy manure.

Subsurface Manure Injection Reduces Surface Transport of Antibiotic Resistance Genes but May Create Antibiotic Resistance Hotspots in Soils.

Compared to surface application, manure subsurface injection reduces surface runoff of nutrients, antibiotic resistant microorganisms, and emerging contaminants. Less is known regarding the impact of both manure application methods on surface transport of antibiotic resistance genes (ARGs) in manure-amended fields. We applied liquid dairy manure to field plots by surface application and subsurface injection and simulated rainfall on the first or seventh day following application. The ARG richness, relative abundance (normalized to 16s rRNA), and ARG profiles in soil and surface runoff were monitored using shotgun metagenomic sequencing. Within 1 day of manure application, compared to unamended soils, soils treated with manure had 32.5-70.5% greater ARG richness and higher relative abundances of sulfonamide (6.5-129%) and tetracycline (752-3766%) resistance genes (p ≤ 0.05). On day 7, soil ARG profiles in the surface-applied plots were similar to, whereas subsurface injection profiles were different from, that of the unamended soils. Forty-six days after manure application, the soil ARG profiles in manure injection slits were 37% more diverse than that of the unamended plots. The abundance of manure-associated ARGs were lower in surface runoff from manure subsurface injected plots and carried a lower resistome risk score in comparison to surface-applied plots. This study demonstrated, for the first time, that although manure subsurface injection reduces ARGs in the runoff, it can create potential long-term hotspots for elevated ARGs within injection slits.

Dairy manure pellets and palm oil mill effluent as alternative nutrient sources in cultivating Sporosarcina pasteurii for calcium carbonate bioprecipitation.

Microbially induced carbonate precipitation (MICP) is a process that hydrolysis urea by microbial urease to fill the pore spaces of soil with induced calcium carbonate (CaCO3 ) precipitates, which eventually results in improved or solidified soil. This research explored the possibility of using dairy manure pellets (DMP) and palm oil mill effluent (POME) as alternative nutrient sources for Sporosarcina pasteurii cultivation and CaCO3 bioprecipitation. Different concentrations (20-80 g l-1 ) of DMP and POME were used to propagate the cells of S. pasteurii under laboratory conditions. The measured CaCO3 contents for MICP soil specimens that were treated with bacterial cultures grown in DMP medium (60%, w/v) was 15·30 ± 0·04 g ml-1 and POME medium (40%, v/v) was 15·49 ± 0·05 g ml-1 after 21 days curing. The scanning electron microscopy showed that soil treated with DMP had rhombohedral structure-like crystals with smooth surfaces, whilst that of POME entailed ring-like cubical formation with rough surfaces Electron dispersive X-ray analysis was able to identify a high mass percentage of chemical element compositions (Ca, C and O), whilst spectrum from Fourier-transform infrared spectroscopy confirmed the vibration peak intensities for CaCO3 . Atomic force microscopy further showed clear topographical differences on the crystal surface structures that were formed around the MICP treated soil samples. These nutrient sources (DMP and POME) showed encouraging potential cultivation mediums to address high costs related to bacterial cultivation and biocementation treatment.

Evaluating the Feasibility of a Low-Field Nuclear Magnetic Resonance (NMR) Sensor for Manure Nutrient Prediction.

Livestock manure is typically applied to fertilize crops, however the accurate determination of manure nutrient composition through a reliable method is important to optimize manure application rates that maximize crop yields and prevent environmental contamination. Existing laboratory methods can be time consuming, expensive, and generally the results are not provided prior to manure application. In this study, the evaluation of a low-field nuclear magnetic resonance (NMR) sensor designated for manure nutrient prediction was assessed. Twenty dairy manure samples were analyzed for total solid (TS), total nitrogen (TN), ammoniacal nitrogen (NH4-N), and total phosphorus (TP) in a certified laboratory and in parallel using the NMR analyzer. The linear regression of NMR prediction versus lab measurements for TS had an R2 value of 0.86 for samples with TS < 8%, and values of 0.94 and 0.98 for TN and NH4-N, respectively, indicating good correlations between NMR prediction and lab measurements. The TP prediction of NMR for all samples agreed with the lab analysis with R2 greater than 0.87. The intra- and inter-sample variations of TP measured by NMR were significantly larger than other parameters suggesting less robustness in TP prediction. The results of this study indicate low-field NMR is a rapid method that has a potential to be utilized as an alternative to laboratory analysis of manure nutrients, however, further investigation is needed before wide application for on farm analysis.

Evaluation of the Immobilization of Coexisting Heavy Metal Ions of Pb2+, Cd2+ , and Zn2+ from Water by Dairy Manure-Derived Biochar: Performance and Reusability.

Heavy metals including Cd, Pb, and Zn are prevalent stormwater and groundwater contaminants derived from natural and human activities, and there is a lack of cost-effective treatment for their removal. Recently, biochar has been increasingly recognized as a promising low-cost sorbent that can be used to remediate heavy metal contaminated water. This study evaluates the immobilization/release performance of dairy manure-derived biochar (DM-BC) as a sustainable material for competitive removal of coexisting heavy metal ions from water and explains the underlying mechanism for regeneration/reusability of biochar. Results showed that the metal ions exhibited competitive removal in the order of Pb2+ ≫ Zn2+ > Cd2+. The pH played a decisive role in influencing metal ion speciation affecting the electrostatic attraction/repulsion and surface complexation. Higher pH led to greater removal for Pb2+ and Cd2+, whereas Zn2+ showed maximum removal at pH ≈ 7.5. Diffuse reflectance infrared spectroscopy, scanning electron microscopy, and X-ray diffraction confirmed the interactions and precipitation reactions of oxygen-containing functional groups (e.g., ─OH, CO32-, and Si─O) as key participants in metal immobilization. Langmuir, Freundlich, and Redlich-Peterson isotherm modeling data showed varied and unique results depending on the metal ion and concentration. The removal kinetics and model fitting showed that the three steps of intraparticle diffusion might be more representative for describing the immobilization processes of metal ions on the external surface and internal pores. In the flow-through columns, DM-BC effectively retained the mixed metal ions of Cd2+, Pb2+, and Zn2+ showing 100% removal for the duration of the column run over three cycles of regeneration and reuse.

On-Farm Anaerobic Digestion of Dairy Manure Reduces the Abundance of Antibiotic Resistance-Associated Gene Targets and the Potential for Plasmid Transfer.

The present study investigated the impact of on-farm anaerobic digestion on the abundance of enteric bacteria, antibiotic resistance-associated gene targets, and the horizontal transfer potential of extended-spectrum ß-lactamase (ESBL) genes. Samples of raw and digested manure were obtained from six commercial dairy farms in Ontario, Canada. Digestion significantly abated populations of viable coliforms in all six farms. Conjugative transfer of plasmids carrying ß-lactamase genes from manure bacteria enriched overnight with buffered peptone containing 4 mg/liter cefotaxime into a ß-lactam-sensitive green fluorescent protein (GFP)-labeled Escherichia coli recipient strain was evaluated in patch matings. Digestion significantly decreased the frequency of the horizontal transfer of ESBL genes. Twenty-five transconjugants were sequenced, revealing six distinct plasmids, ranging in size from 40 to 180 kb. A variety of ESBL genes were identified: blaCTX-M-1, blaCTX-M-14, blaCTX-M-15, blaCTX-M-27, blaCTX-M-55, and blaPER-1. blaCTX-M-15 was the most prevalent ESBL gene detected on plasmids harbored by transconjugants. Various mobile genetic elements were found located proximal to resistance genes. Ten gene targets, including sul1, str(A), str(B), erm(B), erm(F), intI1, aadA, incW, blaPSE, and blaOXA-20, were quantified by quantitative PCR on a subset of 18 raw and 18 digested samples. Most targets were significantly more abundant in raw manure; however, erm(B) and erm(F) targets were more abundant in digested samples. Overall, on-farm digestion of dairy manure abated coliform bacteria, a number of antibiotic resistance-associated gene targets, and the potential for in vitro conjugation of plasmids conferring resistance to extended-spectrum ß-lactams and other classes of antibiotics into E. coli CV601. IMPORTANCE Using livestock manure for fertilization can entrain antibiotic-resistant bacteria into soil. Manure on some dairy farms is anaerobically digested before being land applied. Recommending the widespread implementation of the practice should be founded on understanding the impact of this treatment on various endpoints of human health concern. Although lab-scale anaerobic treatments have shown potential for reducing the abundance of antibiotic resistance genes, there are very few data from commercial farms. Anaerobic digestion of manure on six dairy farms efficiently abated coliform bacteria, E. coli, and a majority of antibiotic resistance-associated gene targets. In addition, the conjugation potential of plasmids carrying ESBL genes into introduced E. coli strain CV601 was reduced. Overall, anaerobic digestion abated coliform bacteria, the genes that they carry, and the potential for ESBL-carrying plasmid transfer.

Rapid removal of nitrate from liquid dairy manure by cationic poly (vinyl alcohol-co-ethylene) nanofiber membrane.

Elevated levels of nitrate in surface water is a leading concern, which impacts human and animal health, and controlling it requires improved and sustainable methods capable of removing nitrate anions from source waste water to reduce nitrate anions transport to environment. In liquid dairy manure, nitrogen content can vary from 200 to 600 ppm and the transport of manure nitrogen into ambient water through hydrologic processes has a potential to exceed the maximum contaminant level limit (10 ppm) of nitrate nitrogen for regulated public water systems. Dairy manure is considered as a reservoir of nitrate. This research investigates on the determination of optimal designing of nanofiber membrane to remove nitrate anions from liquid dairy manure. A cationic poly (vinyl alcohol-co-ethylene) nanofiber membrane (EVOH) NFM was grafted via UV with 2-(methacryloyloxy) ethyl trimethylammonium chloride (DMAC) monomers. The adsorption efficiency of nitrate by the membrane was determined on liquid manure of dairy lagoons located in Central Valley of California. Initial nitrate concentrations in dairy manure varied from 75 to 100 ppm. Results showed that nitrate in dairy water was removed by 70% in 40 min. Tortuous structure and chemical stability of membrane resulted in nitrate dynamic binding capacity of 40 mg g-1. Furthermore, it exhibits efficient reusability without significant changes in its performance using 0.5 M sodium hydroxide solution for nitrate desorption. Results showed that change in pH, and multi-anion conditions had limited effects on nitrate removal efficiency, and EVOH NFM can be a viable option to remove nitrate of liquid manure. This could be used for mitigating transport of excess nitrate from manure to environment. Overall, the results suggest that EVOH-g-DMAC NFM is efficient, low-cost (13 USD/m3) and recyclable material for sustainable removal of nitrate from dairy manure wastewater without requiring any ionic strength or pH adjustment.

Short communication: Increasing temperature and pH can facilitate reductions of cephapirin and antibiotic resistance genes in dairy manure slurries.

Quantifying antibiotics and antibiotic resistance genes (ARG) in manure exposed to various temperature and pH treatments could guide the development of cost-effective manure handling methods to minimize the spread of antibiotic resistance following land application of manure. This study aimed to investigate the effect of various temperatures and initial pH shocks on the persistence of a cephalosporin antibiotic and ARG in dairy manure slurries. Feces and urine were collected from 5 healthy dairy cows administered with cephapirin (cephalosporin antibiotic) at dry-off via intramammary infusion and were mixed with sterile water to generate manure slurries. In a 28-d incubation study, dairy manure slurries either were continuously exposed to 1 of 3 temperatures (10, 35, and 55°C) or received various initial pH (5, 7, 9, and 12) shocks. Cephapirin was detected in the initial samples and on d 1 following all treatments, but it was undetectable thereafter. This indicates that cephapirin can be rapidly degraded irrespective of temperature and pH treatments. However, degradation was greater on d 1 with the mesophilic (35°C) and thermophilic (55°C) environments compared with the psychrophilic environment (10°C). Increasing pH beyond neutral also accelerated degradation as cephapirin concentrations were lower on d 1 after initial alkaline adjustments (pH 9 and 12) than after neutral and acidic adjustments (pH 7 and 5). No significant effect of temperature or initial pH was observed on abundances of a ß-lactam ARG, cfxA, and a tetracycline ARG, tet(W), implying that bacteria that encoded cfxA or tet(W) genes were not sensitive to temperature or pH in dairy manure slurries. However, abundances of a macrolide ARG, mefA, were decreased in the psychrophilic and thermophilic environments and also following exposure to a strong alkaline shock (pH 12). Our results suggest that increasing temperature or pH during storage of dairy manure slurries could be used together with other on-farm practices that are tailored to reduce the transfer of ARG from manure to the environment following land application.

Survival of Escherichia coli in Manure-Amended Soils Is Affected by Spatiotemporal, Agricultural, and Weather Factors in the Mid-Atlantic United States.

Untreated biological soil amendments of animal origin (BSAAO), such as manure, are commonly used to fertilize soils for growing fruit and vegetable crops and can contain enteric bacterial foodborne pathogens. Little is known about the comparative longitudinal survival of pathogens in agricultural fields containing different types of BSAAO, and field data may be useful to determine intervals between manure application and harvest of produce intended for human consumption to minimize foodborne illness. This study generated 324 survival profiles from 12 different field trials at three different sites (UMES, PA, and BARC) in the Mid-Atlantic United States from 2011 to 2015 of inoculated nonpathogenic Escherichia coli (gEc) and attenuated O157 E. coli (attO157) in soils which were unamended (UN) or amended with untreated poultry litter (PL), horse manure (HM), or dairy manure solids (DMS) or liquids (DML). Site, season, inoculum level (low/high), amendment type, management (organic/conventional), and depth (surface/tilled) all significantly (P < 0.0001) influenced survival duration (dpi100mort). Spatiotemporal factors (site, year, and season) in which the field trial was conducted influenced survival durations of gEc and attO157 to a greater extent than weather effects (average daily temperature and rainfall). Initial soil moisture content was the individual factor that accounted for the greatest percentage of variability in survival duration. PL supported greater survival durations of gEc and attO157, followed by HM, UN, and DMS in amended soils. The majority of survival profiles for gEc and attO157 which survived for more than 90 days came from a specific year (i.e., 2013). The effect of management and depth on dpi100mort were dependent on the amendment type evaluated.IMPORTANCE Current language in the Food Safety Modernization Act Produce Safety Rule states no objection to a 90- or 120-day interval between application of untreated BSAAO and harvest of crops to minimize transfer of pathogens to produce intended for human consumption with the intent to limit potential cases of foodborne illness. This regional multiple season, multiple location field trial determined survival durations of Escherichia coli in soils amended with manure to determine whether this interval is appropriate. Spatiotemporal factors influence survival durations of E. coli more than amendment type, total amount of E. coli present, organic or conventional soil management, and depth of manure application. Overall, these data show poultry litter may support extended survival of E. coli compared to horse manure or dairy manure, but spatiotemporal factors like site and season may have more influence than manure type in supporting survival of E. coli beyond 90 days in amended soils in the Mid-Atlantic United States.

Composts of poultry litter or dairy manure differentially affect survival of enteric bacteria in fields with spinach.

AIMS: The aim was to determine the survival and persistence of Escherichia coli in soil amended with compost from different manure sources. METHOD AND RESULTS: Complex interactions of abiotic and biotic factors on E. coli survival were characterized in field experiment plots receiving randomly assigned compost treatments: dairy windrow, dairy vermicompost, poultry windrow or no compost. Biomass, activity and function of indigenous microbial communities in the composts and soils were measured concurrently to determine whether mechanisms of compost were driven by biotic or abiotic properties. E. coli persisted in compost containing poultry amendments but not in composts containing dairy or no amendments. Poultry compost contained more NH4 -N and a distinct microbial community compared to dairy and no compost treatments. A laboratory experiment performed on compost extracts suggested that E. coli survived better in extracts devoid of indigenous microbes as long as bioavailable nutrients were plentiful. CONCLUSIONS: Dairy-based composts are less likely to support E. coli survival than poultry-based composts. SIGNIFICANCE AND IMPACT OF THE STUDY: Results aid in risk assessment of the use of different types of manure-based compost and soil amendments in fruit and vegetable production by elucidating the roles of nutrient and microbial community composition on survival of E. coli in amended field soils.

Bacterial community succession in dairy manure composting with a static composting technique.

This study applied high-throughput sequencing technology and PICRUSt (phylogenetic investigation of communities by reconstruction of unobserved state) to examine the microbial population dynamics during the composting of dairy manure and rice straw in a static (without turning) composting system. The results showed that the composition of the bacterial community varied significantly during the composting process. The dominant phyla included Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Chloroflexi. Biodiversity indices showed that bacterial community diversity had the peak value during the mesophilic phase. Redundancy analysis indicated that nitrogen was the most important factor in the distribution of genera during the composting process. Finally, the Pearson correlation results suggested that Thermomonospora and Thermopolyspora could be the biomarkers of the composting maturation phase. The metabolic characteristics of the bacterial communities were predicted by PICRUSt. The result showed that metabolism of amino acids, lipids, and most of the carbohydrates increased during the whole composting treatment. However, methane metabolism, carbon fixation pathways in prokaryotes, and nucleotide metabolism decreased after the thermophilic phase. The present study provides a better understanding for bacterial community composition and function succession in dairy manure composting.

Targeting Bacteria and Methanogens To Understand the Role of Residual Slurry as an Inoculant in Stored Liquid Dairy Manure.

Microbial communities in residual slurry left after removal of stored liquid dairy manure have been presumed to increase methane emission during new storage, but these microbes have not been studied. While actual manure storage tanks are filled gradually, pilot- and farm-scale studies on methane emissions from such systems often use a batch approach. In this study, six pilot-scale outdoor storage tanks with (10% and 20%) and without residual slurry were filled (gradually or in batch) with fresh dairy manure, and methane and methanogenic and bacterial communities were studied during 120 days of storage. Regardless of filling type, increased residual slurry levels resulted in higher abundance of methanogens and bacteria after 65 days of storage. However, stronger correlation between methanogen abundance and methane flux was observed in gradually filled tanks. Despite some variations in the diversity of methanogens or bacteria with the presence of residual slurry, core phylotypes were not impacted. In all samples, the phylum Firmicutes predominated (∼57 to 70%) bacteria: >90% were members of ClostridiaMethanocorpusculum dominated (∼57 to 88%) archaeal phylotypes, while Methanosarcina gradually increased with storage time. During peak flux of methane, Methanosarcina was the major player in methane production. The results suggest that increased levels of residual slurry have little impact on the dominant methanogenic or bacterial phylotypes, but large population sizes of these organisms may result in increased methane flux during the initial phases of storage.IMPORTANCE Methane is the major greenhouse gas emitted from stored liquid dairy manure. Residual slurry left after removal of stored manure from tanks has been implicated in increasing methane emissions in new storages, and well-adapted microbial communities in it are the drivers of the increase. Linking methane flux to the abundance, diversity, and activity of microbial communities in stored slurries with different levels of residual slurry can help to improve the mitigation strategy. Mesoscale and lab-scale studies conducted so far on methane flux from manure storage systems used batch-filled tanks, while the actual condition in many farms involves gradual filling. Hence, this study provides important information toward determining levels of residual slurry that result in significant reduction of well-adapted microbial communities prior to storage, thereby reducing methane emissions from manure storage tanks filled under farm conditions.

Redundancy in Anaerobic Digestion Microbiomes during Disturbances by the Antibiotic Monensin.

The antibiotic monensin is fed to dairy cows to increase milk production efficiency. A fraction of this monensin is excreted into the cow manure. Previous studies have found that cow manure containing monensin can negatively impact the performance of anaerobic digesters, especially upon first introduction. Few studies have examined whether the anaerobic digester microbiome can adapt to monensin during the operating time. Here, we conducted a long-term time series study of four lab-scale anaerobic digesters fed with cow manure. We examined changes in both the microbiome composition and function of the anaerobic digesters when subjected to the dairy antibiotic monensin. In our digesters, monensin was not rapidly degraded under anaerobic conditions. The two anaerobic digesters that were subjected to manure from monensin feed-dosed cows exhibited relatively small changes in microbiome composition and function due to relatively low monensin concentrations. At higher concentrations of monensin, which we dosed directly to control manure (from dairy cows without monensin), we observed major changes in the microbiome composition and function of two anaerobic digesters. A rapid introduction of monensin to one of these anaerobic digesters led to the impairment of methane production. Conversely, more gradual additions of the same concentrations of monensin to the other anaerobic digester led to the adaptation of the anaerobic digester microbiomes to the relatively high monensin concentrations. A member of the candidate OP11 (Microgenomates) phylum arose in this anaerobic digester and appeared to be redundant with certain Bacteroidetes phylum members, which previously were dominating.IMPORTANCE Monensin is a common antibiotic given to dairy cows in the United States and is partly excreted with dairy manure. An improved understanding of how monensin affects the anaerobic digester microbiome composition and function is important to prevent process failure for farm-based anaerobic digesters. This time series study demonstrates how anaerobic digester microbiomes are inert to low monensin concentrations and can adapt to relatively high monensin concentrations by redundancy in an already existing population. Therefore, our work provides further insight into the importance of microbiome redundancy in maintaining the stability of anaerobic digesters.