Comparison and Evaluation of GHG Emissions during Simulated Thermophilic Composting of Different Municipal and Agricultural Feedstocks.

Composting is widely used to recycle a variety of different organic wastes. In this study, dairy manure, chicken litter, biosolids, yard trimmings and food waste were selected as representative municipal and agricultural feedstocks and composted in simulated thermophilic composting reactors to compare and evaluate the GHG emissions. The results showed that the highest cumulative emissions of CO2, CH4 and N2O were observed during yard trimmings composting (659.14 g CO2 kg-1 DM), food waste composting (3308.85 mg CH4 kg-1 DM) and chicken litter composting (1203.92 mg N2O kg-1 DM), respectively. The majority of the carbon was lost in the form of CO2. The highest carbon loss by CO2 and CH4 emissions and the highest nitrogen loss by N2O emission occurred in dairy manure (41.41%), food waste (0.55%) and chicken litter composting (3.13%), respectively. The total GHG emission equivalent was highest during food waste composting (365.28 kg CO2-eq ton-1 DM) which generated the highest CH4 emission and second highest N2O emissions, followed by chicken litter composting (341.27 kg CO2-eq ton-1 DM), which had the highest N2O emissions. The results indicated that accounting for GHG emissions from composting processes when it is being considered as a sustainable waste management practice was of great importance.

Control of dimethyl sulfide and dimethyl disulfide odors during pig manure composting using nitrogen amendment.

Effects of nitrogen electron acceptors on dimethyl sulfide (Me2S) and dimethyl disulfide (Me2SS) odor emission during composting was investigated. The chemicals and doses used included sodium nitrate (NO3- at 10mM, 20mM and 40mM), sodium nitrite (NO2- at 10mM, 20mM and 40mM) and sodium nitrite (10mM, 20mM and 40mM) with hexaammonium heptamolybdate tetrahydrate (HHT). The results showed that the addition of these chemicals restricted the emission of Me2S and Me2SS. The emission reduction effect of NO2- was greater than NO3- at the same dosage. A greater reduction was observed when HHT was also added. With 2mM HHT+40mM NO2- addition, the emission of Me2S and Me2SS was reduced by 92.3% and 82.3%, respectively. Comparison of compost maturity indices for treated and untreated composts indicated that none of the additives adversely affected compost quality. These results indicate that nitrogen chemical addition may provide an efficient method to control sulfur odors during composting.

Comparison of solid-state anaerobic digestion and composting of yard trimmings with effluent from liquid anaerobic digestion.

Solid-state anaerobic digestion (SS-AD) and composting of yard trimmings with effluent from liquid AD were compared under thermophilic condition. Total solids (TS) contents of 22%, 25%, and 30% were studied for SS-AD, and 35%, 45%, and 55% for composting. Feedstock/effluent (F/E) ratios of 2, 3, 4, 5, and 6 were tested. In composting, the greatest carbon loss was obtained at 35% TS, which was 2-3 times of that at 55% TS and was up to 50% higher than that in SS-AD. In SS-AD, over half of the degraded carbon was converted to methane with the greatest methane yield of 121 L/kg VS(feedstock). Methane production from SS-AD was low at F/E ratios of 2 and 3, likely due to the inhibitory effect of high concentrations of ammonia nitrogen (up to 5.6g/kg). The N-P-K values were similar for SS-AD digestate and compost with different dominant nitrogen forms.

Composting of waste paint sludge containing melamine resin and the compost's effect on vegetable growth and soil water quality.

Melamine resin (MR) is introduced to the environment from many industrial effluents, including waste paint sludge (WPS) from the automobile industry. Melamine resin contains a high nitrogen (N) content and is a potential N source during composting. In this study, two carbon sources, waste paper (WP) and plant residue (PR), were used to study their effects on composting of WPS. Additional work tested the WPS-composts effects on plant growth and soil water quality. After 84 days of composting, 85% and 54% of the initial MR was degraded in WP- and PR-composts, respectively. The limiting factor was that the MR created clumps during composting so that decomposition was slowed. Compared to the untreated control, both WP- and PR-composts increased growth of cucumber (Cucumis sativus), radish (Raphanus sativus) and lettuce (Lactuca sativa). Concentrations of trace elements in plants and soil water did not rise to a level that would preclude WPS-composts from being used as a soil amendment.

Composting of waste paint sludge containing melamine resin as affected by nutrients and gypsum addition and microbial inoculation.

Melamine formaldehyde resins have hard and durable properties and are found in many products, including automobile paints. These resins contain high concentrations of nitrogen and, if properly composted, can yield valuable products. We evaluated the effects of starter compost, nutrients, gypsum and microbial inoculation on composting of paint sludge containing melamine resin. A bench-scale composting experiment was conducted at 55 °C for 91 days and then at 30 °C for an additional 56 days. After 91 days, the composts were inoculated with a mixed population of melamine-degrading microorganisms. Melamine resin degradation after the entire 147 days of composting varied between 73 and 95% for the treatments with inoculation of microorganisms compared to 55-74% for the treatments without inoculation. Degradation was also enhanced by nutrients and gypsum additions. Our results infer that large scale composting of melamine resins in paint sludge is possible.

Persistence of resistance to erythromycin and tetracycline in swine manure during simulated composting and lagoon treatments.

The use of antimicrobials in food animal production leads to the development of antimicrobial resistance (AMR), and animal manure constitutes the largest reservoir of such AMR. In previous studies, composted swine manure was found to contain substantially lower abundance of AMR genes that encode resistance to tetracyclines (tet genes) and macrolide-lincosamide-streptogramin B (MLS(B)) superfamily (erm genes), than manures that were treated by lagoons or biofilters. In this study, temporal changes in AMR carried by both cultivated and uncultivated bacteria present in swine manure during simulated composting and lagoon storage were analyzed. Treatments were designed to simulate the environmental conditions of composting (55°C with modest aeration) and lagoon storage (ambient temperature with modest aeration). As determined by selective plate counting, over a 48-day period, cultivated aerobic heterotrophic erythromycin-resistant bacteria and tetracycline-resistant bacteria decreased by more than 4 and 7 logs, respectively, in the simulated composting treatment while only 1 to 2 logs for both resistant bacterial groups in the simulated lagoon treatment. Among six classes each of erm and tet genes quantified by class-specific real-time PCR assays, the abundance of erm(A), erm(C), erm(F), erm(T), erm(X), tet(G), tet(M), tet(O), tet(T), and tet(W) declined marginally during the first 17 days, but dramatically thereafter within 31 days of the composting treatment. No appreciable reduction of any of the erm or tet genes analyzed was observed during the simulated lagoon treatment. Correlation analysis showed that most of the AMR gene classes had similar persistence pattern over the course of the treatments, though not all AMR genes were destructed at the same rate during the treatments.

Effects of organic loading rate on reactor performance and archaeal community structure in mesophilic anaerobic digesters treating municipal sewage sludge.

In this study, the organic loading rate (OLR) of a high-solids anaerobic digestion (HSAD) system was increased from 3.4 to 5.0 gVS L(-1) day(-1) and reactor stability, performance and microbial community structure were determined. Laboratory simulations (3.5 L) of the full-scale process (500 dry ton year(-1)) were conducted using continuously stirred-tank mesophilic reactors. OLRs of 3.4 gVS L(-1)day(-1) (equal to the full-scale HSAD), 4.0, 4.5 and 5.0 gVS L(-1)day(-1) were evaluated. Biochemical parameters and archaeal community dynamics were measured over 42 days of steady state operation. Results showed that increasing OLR increased the amount of organic matter conversion and resulted in higher organic matter removal and volumetric methane (CH4) production (VMP) rates. The highest volatile solids (VS) removal and VMP results of 54 ± 2% and 1.4 ± 0.1 L CH4 L(-1)day(-1) were observed for 5.0 gVS L(-1) day(-1). The efficiency of reactor conversion of organic matter to CH(4) was found to be similar in all the treatments with an average value of 0.57 ± 0.07 LCH(4) gVS(-1) (removed). 16S rRNA gene terminal restriction fragment polymorphism (T-RFLP) analyses revealed that archaeal TRFs remained stable during the experiment accounting for an average relative abundance (RA) of 81 ± 1%. Archaea consistent with multiple terminal restriction fragments (TRFs) included members of the Euryarchaeota and Crenarchaeota phyla, including acetoclastic and hydrogenotrophic groups. In conclusion, this laboratory-scale study suggests that performance and stability as well as the archaeal community structure in this HSAD system was unaffected by increasing the OLR by nearly 50% and that this increase resulted in a similar increase in the amount of CH(4) gas generated.

Effects of different wavelengths of light on lignin peroxidase production by the white-rot fungi Phanerochaete chrysosporium grown in submerged cultures.

In this study, the effects of different wavelengths of light (UV, blue, green, yellow, red) and white light on lignin peroxidase (LiP), protein, biomass and exo-polysaccharide production and glucose uptake by Phanerochaete chrysosporium BKM-F-1767 were determined. The experiments were conducted under aerated (CS) and oxygenated (RS) culture conditions. The results showed that only green light significantly increased maximum LiP production (by 20% and 27% in CS and RS cultures respectively). Green light also increased biomass production in oxygenated cultures (RS). Blue and UV light both significantly reduced maximum LiP activity. Yellow, red and white lights had mixed effects on culture properties. This is the first time that the effects of different wavelengths of light on lignin peroxidase production and other culture properties have been investigated. The novel findings may be important in improving the yield of lignin modifying enzymes for biomass conversion processes and understanding their regulation.

Occurrence and persistence of erythromycin resistance genes (erm) and tetracycline resistance genes (tet) in waste treatment systems on swine farms.

Animal manure from modern animal agriculture constitutes the single largest source of antibiotic resistance (AR) owing to the use of large quantities of antibiotics. After animal manure enters the environment, the AR disseminates into the environment and can pose a potentially serious threat to the health and well-being of both humans and animals. In this study, we evaluated the efficiency of three different on-farm waste treatment systems in reducing AR. Three classes of erythromycin resistance genes (erm) genes (B, F, and X) conferring resistances to macrolide-lincosamides-streptogramin B (MLS(B)) and one class of tetracycline resistance genes (tet) gene (G) conferring resistance to tetracyclines were used as models. Real-time polymerase chain reaction assays were used to determine the reservoir sizes of these AR genes present in the entire microbiome. These classes of AR genes varied considerably in abundance, with erm(B) being more predominant than erm(F), erm(X), and tet(G). These AR genes also varied in persistence in different waste treatment systems. Aerobic biofiltration reduced erm(X) more effectively than other AR genes, while mesophilic anaerobic digestion and lagoon storage did not appreciably reduce any of these AR genes. Unlike chemical pollutants, some AR genes could increase after reduction in a preceding stage of the treatment processes. Season might also affect the persistence of AR. These results indicate that AR arising from swine-feeding operations can survive typical swine waste treatment processes and thus treatments that are more effective in destructing AR on farms are required.

Contrasting patterns of seed and root colonization by bacteria from the genus Chryseobacterium and from the family Oxalobacteraceae.

Microbial colonization of plant seeds and roots is a highly complex process in which soil and plant type can influence the composition of the root-associated and rhizosphere microbial communities. Amendment of compost, a common agricultural technique, introduces exogenous nutrients and microorganisms to the soil-plant environment, and can further influence microbial community composition in the plant environment. Although compost amendments can strongly influence soil and rhizosphere microbial communities, there is evidence that with increasing proximity to the root, plant influences predominate over soil effects. We hypothesized that the 'rhizosphere effect' observed with proximity to plant surfaces does not act equally on all microorganisms. To explore this issue, we examined two bacterial taxa that reproducibly colonized seed and root surfaces in an experiment examining the influence of compost amendment on plant-associated bacterial communities. Population-specific analyses revealed striking differences in the ecology of bacteria from the genus Chryseobacterium and the family Oxalobacteraceae in potting mix and plant-associated environments. Seed- and root-colonizing Oxalobacteraceae populations were highly sensitive to plant effects, and phylogenetic analyses of root-colonizing Oxalobacteraceae revealed the presence of root-associated populations that were highly similar, regardless of treatment, and differed from the potting mix populations detected at the same sampling points. Conversely, Chryseobacterium community composition was found to be essentially invariant within treatments, but was strongly influenced by compost amendment. This persistence and stable nature of the Chryseobacterium community composition demonstrates that rhizosphere selection is not the exclusive factor involved in determining the composition of the cucumber spermosphere and rhizosphere communities.

Effects of different animal waste treatment technologies on detection and viability of porcine enteric viruses.

Enteric pathogens in animal waste that is not properly processed can contaminate the environment and food. The persistence of pathogens in animal waste depends upon the waste treatment technology, but little is known about persistence of porcine viruses. Our objectives were to characterize the porcine enteric viruses (porcine noroviruses [PoNoVs], porcine sapoviruses [PoSaVs], rotavirus A [RV-A], RV-B, and RV-C) in fresh feces or manure and to evaluate the effects of different candidate environmentally superior technologies (ESTs) for animal waste treatment on the detection of these viruses. Untreated manure and samples collected at different stages during and after treatment were obtained from swine farms that used conventional waste management (CWM) and five different candidate ESTs. The RNA from porcine enteric viruses was detected by reverse transcription-PCR and/or seminested PCR; PoSaV and RV-A were also detected by enzyme-linked immunosorbent assay. Cell culture immunofluorescence (CCIF) and experimental inoculation of gnotobiotic (Gn) pigs were used to determine RV-A/C infectivity in posttreatment samples. The PoSaV and RV-A were detected in pretreatment samples from each farm, whereas PoNoV and RV-C were detected in pretreatment feces from three of five and four of five farms using the candidate ESTs, respectively. After treatment, PoSaV RNA was detected only in the samples from the farm using CWM and not from the farms using the candidate ESTs. RV-A and RV-C RNAs were detected in four of five and three of four candidate ESTs, respectively, after treatment, but infectious particles were not detected by CCIF, nor were clinical signs or seroconversion detected in inoculated Gn pigs. These results indicate that only RV-A/C RNA, but no viral infectivity, was detected after treatment. Our findings address a public health concern regarding environmental quality surrounding swine production units.

Development and application of real-time PCR assays for quantification of erm genes conferring resistance to macrolides-lincosamides-streptogramin B in livestock manure and manure management systems.

Erythromycin and tylosin are commonly used in animal production, and such use is perceived to contribute to the overall antimicrobial resistance (AR) reservoirs. Quantitative measurements of this type of AR reservoir in microbial communities are required to understand AR ecology (e.g., emergence, persistence, and dissemination). We report here the development, validation, and use of six real-time PCR assays for quantifying six classes of erm genes (classes A through C, F, T, and X) that encode the major mechanism of resistance to macrolides-lincosamides-streptogramin B (MLS(B)). These real-time PCR assays were validated and used in quantifying the six erm classes in five types of samples, including those from bovine manure, swine manure, compost of swine manure, swine waste lagoons, and an Ekokan upflow biofilter system treating hog house effluents. The bovine manure samples were found to contain much smaller reservoirs of each of the six erm classes than the swine manure samples. Compared to the swine manure samples, the composted swine manure samples had substantially reduced erm gene abundances (by up to 7.3 logs), whereas the lagoon or the biofilter samples had similar erm gene abundances. These preliminary results suggest that the methods of manure storage and treatment probably have a substantial impact on the persistence and decline of MLS(B) resistance originating from food animals, thus likely affecting the dissemination of such resistance genes into the environment. The abundances of these erm genes appeared to be positively correlated with those of the tet genes determined previously among these samples. These real-time PCR assays provide a rapid, quantitative, and cultivation-independent measurement of six major classes of erm genes, which should be useful for ecological studies of AR.

Succession of bacterial communities during early plant development: transition from seed to root and effect of compost amendment.

Compost amendments to soils and potting mixes are routinely applied to improve soil fertility and plant growth and health. These amendments, which contain high levels of organic matter and microbial cells, can influence microbial communities associated with plants grown in such soils. The purpose of this study was to follow the bacterial community compositions of seed and subsequent root surfaces in the presence and absence of compost in the potting mix. The bacterial community compositions of potting mixes, seed, and root surfaces sampled at three stages of plant growth were analyzed via general and newly developed Bacteroidetes-specific, PCR-denaturing gradient gel electrophoresis methodologies. These analyses revealed that seed surfaces were colonized primarily by populations detected in the initial potting mixes, many of which were not detected in subsequent root analyses. The most persistent bacterial populations detected in this study belonged to the genus Chryseobacterium (Bacteroidetes) and the family Oxalobacteraceae (Betaproteobacteria). The patterns of colonization by populations within these taxa differed significantly and may reflect differences in the physiology of these organisms. Overall, analyses of bacterial community composition revealed a surprising prevalence and diversity of Bacteroidetes in all treatments.

Persistence of Mycobacterium avium subsp. paratuberculosis and other zoonotic pathogens during simulated composting, manure packing, and liquid storage of dairy manure.

Livestock manures contain numerous microorganisms which can infect humans and/or animals, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis). The effects of commonly used manure treatments on the persistence of these pathogens have rarely been compared. The objective of this study was to compare the persistence of artificially inoculated M. paratuberculosis, as well as other naturally occurring pathogens, during the treatment of dairy manure under conditions that simulate three commonly used manure management methods: thermophilic composting at 55 degrees C, manure packing at 25 degrees C (or low-temperature composting), and liquid lagoon storage. Straw and sawdust amendments used for composting and packing were also compared. Manure was obtained from a large Ohio free-stall dairy herd and was inoculated with M. paratuberculosis at 10(6) CFU/g in the final mixes. For compost and pack treatments, this manure was amended with sawdust or straw to provide an optimal moisture content (60%) for composting for 56 days. To simulate liquid storage, water was added to the manure (to simulate liquid flushing and storage) and the slurry was placed in triplicate covered 4-liter Erlenmeyer flasks, incubated under ambient conditions for 175 days. The treatments were sampled on days 0, 3, 7, 14, 28, and 56 for the detection of pathogens. The persistence of M. paratuberculosis was also assessed by a PCR hybridization assay. After 56 days of composting, from 45 to 60% of the carbon in the compost treatments was converted to CO2, while no significant change in carbon content was observed in the liquid slurry. Escherichia coli, Salmonella, and Listeria were all detected in the manure and all of the treatments on day 0. After 3 days of composting at 55 degrees C, none of these organisms were detectable. In liquid manure and pack treatments, some of these microorganisms were detectable up to 28 days. M. paratuberculosis was detected by standard culture only on day 0 in all the treatments, but was undetectable in any treatment at 3 and 7 days. On days 14, 28, and 56, M. paratuberculosis was detected in the liquid storage treatment but remained undetectable in the compost and pack treatments. However, M. paratuberculosis DNA was detectable through day 56 in all treatments and up to day 175 in liquid storage treatments. Taken together, the results indicate that high-temperature composting is more effective than pack storage or liquid storage of manure in reducing these pathogens in dairy manure. Therefore, thermophilic composting is recommended for treatment of manures destined for pathogen-sensitive environments such as those for vegetable production, residential gardening, or application to rapidly draining fields.

Development and application of real-time PCR assays for quantification of genes encoding tetracycline resistance.

We report here the development, validation, and use of three real-time PCR assays to quantify the abundance of the following three groups of tetracycline resistance genes: tet(A) and tet(C); tet(G); and tet genes encoding ribosomal protection proteins, including tet(M), tet(O), tetB(P), tet(Q), tet(S), tet(T), and tet(W). The assays were validated using known numbers of sample-derived tet gene templates added to microbiome DNA. These assays are both precise and accurate over at least 6 log tet gene copies. New tet gene variants were also identified from cloned tet amplicons as part of this study. The utility of these real-time PCR assays was demonstrated by quantifying the three tet gene groups present in bovine and swine manures, composts of swine manure, lagoons of hog house effluent, and samples from an Ekokan upflow biofilter system treating hog house effluent. The bovine manures were found to contain fewer copies of all three groups of tet genes than the swine manures. The composts of swine manures had substantially reduced tet gene abundance (up to 6 log), while lagoon storage or the upflow biofilter had little effect on tet gene abundance. These results suggest that the method of manure storage and treatment may have a substantial impact on the persistence and dissemination of tet genes in agricultural environments. These real-time PCR assays provide rapid, quantitative, cultivation-independent measurements of 10 major classes of tet genes, which should be useful for ecological studies of antibiotic resistance.

Similarity of bacterial communities in sawdust- and straw-amended cow manure composts.

We analyzed bacterial communities in two cow manure composts derived from the same feed manure and composted in the same location, but composted with different carbon amendments, and in peat-based potting mixes amended with these composts. Bacterial communities were characterized by PCR-denaturing gradient gel electrophoresis (DGGE) analysis of extracted DNAs, and population fingerprints generated for each sample were compared. Sequence analyses of dominant DGGE bands revealed that members of the phylum Bacteroidetes were the most dominant bacteria detected in this study (19 of 31 clones). These analyses demonstrate that bacterial community profiles of individual composts were highly similar, as were profiles of compost-amended potting mixes. However, potting mix profiles differed substantially from the original compost profiles and from that of the peat base. These data indicate that highly similar bacterial populations were active in the two composts, and suggest that the effects of the initial carbon amendment on the mature compost bacterial communities were minor, while factors such as the feed manure and composting location may have been more influential.

Evaluation of two recovery methods for detection of Mycobacterium avium subsp. paratuberculosis by PCR: direct-dilution--centrifugation and C(18)-carboxypropylbetaine processing.

A duplex polymerase chain reaction (PCR)-hybridization assay based on Mycobacterium avium subsp. paratuberculosis (MAP)-specific IS900 integration sites was used to evaluate two mycobacterial recovery methods from bovine feces: a direct-dilution-centrifugation method and a C(18)-carboxypropylbetaine (CB-18)-based method. All MAP PCR results were confirmed for absence of inhibitors using a novel PCR system based on the rpoB gene of plant chloroplasts as an internal control. The detection limits of both MAP recovery methods when coupled with PCR were determined to be between 100 and 1000 organisms. Using culture as a 'gold standard' PCR following the direct-dilution-centrifugation protocol was 92.6% sensitive and 83.7% specific, whereas PCR following the CB-18 method was 100% sensitive and 53.5% specific. Both methods were 100% specific when 60 'true' negatives from two uninfected herds were tested. Both the CB-18 and direct processing methods coupled with a target-specific amplification technique may provide greater sensitivity to diagnose subclinical animals as they were able to detect more positives, on samples derived from infected herds, than conventional culture methods; however, more extensive investigation and follow-up of suspect animals will be required to fully validate the MAP recovery and molecular detection protocols described.