Influence of select bioenergy by-products on soil carbon and microbial activity: A laboratory study.

Concerns about the negative impacts of crop biomass removal on soil ecological functions have led to questioning the long-term sustainability of bioenergy production. To offset this potential negative impact, use of organic C rich by-products from the bioenergy industries have been proposed as a means to replenish soil C in degraded soils. However, the impact of these by-products application on soil carbon dynamics is not fully understood. We measured biogeochemical changes in soil organic C following a three-year field application of two by-products, biochar (BC) and fermentation-by product (FBP), of bioenergy industry processes in an elephant grass [Pennisetum purpureum (L.) Schum.] field. There was a significant increase in overall soil organic C (SOC) observed in BC (270%) treated plots, however the higher labile SOC (51%) content was present in FBP treated plots. Solid-state 13C NMR spectroscopy further revealed increased aromatic and alkyl groups in BC amended soils which lend to its significantly higher hydrophobicity index, HI (2.13) compared with FBP amended soils (HI = 0.8). Initial biogeochemical responses of amended soils to drought conditions were also investigated during a short-term experiment with drying and rewetting of soils. Increased concentrations of extractable C and higher stimulation of microbial activities (respiration and enzyme activities) in FBP amended soils were measured. Overall, our results reveal different impacts of the two soil amendments, where FBP soil application can affect the labile SOC availability, and stimulate rapid microbial response in drought affected soils, and biochar soil application lowers the labile SOC and microbial stimulation facilitating C sequestration over time.

Adsorption of viruses to soil: impact of anaerobic treatment.

The adsorption of viruses in untreated flushed dairy manure wastewater (FDMW), anaerobically digested flushed dairy manure wastewater (ADFDMW) and groundwater to sandy soil was investigated. Batch adsorption studies showed differential adsorption of viruses in groundwater to soil. Less than 75% of PRD1 and MS2 added to groundwater adsorbed after 1 h, but greater than 95% of phiX174 and poliovirus 1 adsorbed to the soil. Adsorption differences in groundwater were related to the isoelectric points of the viruses. Suspending phages in untreated and treated wastewater reduced adsorption compared with groundwater. For MS2, more phages were adsorbed using ADFDMW than with FDMW. Adsorption of poliovirus 1 was not affected by FDMW and ADFDMW. Small column studies (6 x 2.5 cm) produced a similar trend in that adsorption was observed with groundwater and both FDMW and ADFDMW reduced virus adsorption. Groundwater, FDMW or ADFDMW did not affect the adsorption of poliovirus 1 in column studies. The major difference between FDMW and ADFDMW was in mobilisation of adsorbed viruses. The application of FDMW to soil columns with adsorbed viruses caused significantly more viruses to be mobilised than did the application of rainwater or ADFDMW. These results showed that treating FDMW by anaerobic digestion increased the adsorption of viruses to soil and decreased detachment of adsorbed viruses. As the potential for new zoonotic pathogens becomes known, the treatment of animal wastes may become mandatory. The assessment and management of viruses in manure for addressing possible risk to animal and human health is of interest.

Selective growth of Staphylococcus aureus from flushed dairy manure wastewater using acriflavine-supplemented mannitol salt agar.

AIMS: To investigate the use of mannitol salt agar (MSA) supplemented with acriflavine for selective growth and quantification of Staphylococcus aureus from flushed dairy manure wastewater (FDMW). METHODS AND RESULTS: Minimal inhibitory concentrations of acriflavine in MSA were determined by comparing the growth of S. aureus subsp. aureus (ATCC 33591) and Staphylococcus epidermidis (ATCC 155) in pure culture. Acriflavine concentrations of 1.3, 1.4 and 1.5 mg l(-1) reduced CFU of S. epidermidis by 43%, 55% and 87%, respectively, while CFU of S. aureus subsp. aureus were only reduced by 15%, 20% and 26% at the respective concentrations of acriflavine. MSA supplemented with 1.5 mg l(-1) acriflavine was tested for selective growth of indigenous S. aureus from three grab samples of FDMW. Acriflavine concentrations of 1.5 mg l(-1) reduced background flora without significantly reducing (P < 0.05) indigenous S. aureus counts. CONCLUSIONS: Acriflavine-supplemented MSA provides an effective media for selective growth and quantification of indigenous S. aureus from FDMW in the presence of high levels of background microflora. SIGNIFICANCE AND IMPACT OF THE STUDY: S. aureus is implicated for mastitis infections in dairy cows. Therefore, a reliable means for monitoring and detecting the organism in FDMW provides a tool for measuring the effectiveness of treatment for reducing S. aureus levels and implementing flushwater recycling without affecting herd health.

Optimization of low-cost phosphorus removal from wastewater using co-treatments with constructed wetlands.

Eighteen wastewater treatment systems were operated for one year to investigate phosphorus (P) removal. Systems paired co-treatment reactors containing iron or calcium drinking water treatment residuals with vertical-flow constructed wetland mesocosms planted with Schoenoplectus tabernaemontani (K.C. Gmel.) Palla. For secondary municipal wastewater, soluble reactive P (SRP) concentrations were reduced from 0.70 to 0.03 mg L(-1) (95%) or 0.01 mg L(-1) (98%) by systems with the calcium or iron co-treatments, respectively (compared to 0.09 mg L(-1) or 87% by controls). Total P (TP) concentrations were reduced from 1.00 to 0.07 mg L(-1) (93%) and 0.05 mg L(-1) (95%) by the same treatments (compared to 0.16 mg L(-1) or 84% by controls). For anaerobically digested dairy wastewater, SRP was reduced from 7.68 to 6.43 mg L(-1) (16%) or 5.95 mg L(-1) (22%) by the systems with calcium or iron, respectively (compared to 7.37 mg L(-1) or 4% by controls). For this wastewater, the TP was reduced from 48.5 to 22.5 mg L(-1) (53%) and 22.7 mg L(-1) (53%) by the same treatments (compared to 24.1 mg L(-1) or 50% by controls) but performance improved substantially with a design modification tested.

Effects of anaerobic digestion and additives to effluent or cattle feed on odor and odorant concentrations.

Odor intensity (5,437 observations), determined by human panelists (100 different panelists over the course of the experiment), and a number of chemical odorant concentrations were determined for manure-related samples (326) obtained from effluents from conventional stirred-tank reactor (CSTR) and fixed-film anaerobic digesters, effluents to which commercial additives or KMnO4 or H2O2 were added, and feces, urine, and mixed manure from cows fed a control or additive-containing diet. Mostly, samples were held in stoppered, Erlenmeyer flasks for 3 d at room temperature before evaluation by panelists and with chemical analyses, but shorter holding times also were tested. Anaerobic digestion reduced odor intensity linearly with increasing hydraulic retention time (HRT) up to 20 d; fixed-film digestion with 1.5- or 2.3-d HRT reduced odor intensity similarly to that observed with 10-d HRT in CSTR. Addition of commercial products and chemicals altered some odorant concentrations (e.g., ammonia) but did not reduce odor intensity; some products increased odor intensity. Addition of a commercial yeast-based product to a dairy cow diet had no detectable effect. The cow diet study showed that fresh urine and feces alone were less odorous than a mixed combination (manure). Fresh manure was less odorous than manure held for 3 d. Total phenol was the odorant most highly correlated with odor intensity. Individual and total volatile fatty acids also contributed. Ammonia did not seem to be a major contributor to odor in this data set.

Direct isolation of functional genes encoding cellulases from the microbial consortia in a thermophilic, anaerobic digester maintained on lignocellulose.

Gene libraries ("zoolibraries") were constructed in Escherichia coli using DNA isolated from the mixed liquor of thermophilic, anaerobic digesters, which were in continuous operation with lignocellulosic feedstocks for over 10 years. Clones expressing cellulase and xylosidase were readily recovered from these libraries. Four clones that hydrolyzed carboxymethylcellulose and methylumbelliferyl-beta-D-cellobiopyranoside were characterized. All four cellulases exhibited temperature optima (60-65 degrees C) and pH optima (pH 6-7) in accordance with conditions of the enrichment. The DNA sequence of the insert in one clone (plasmid pFGH1) was determined. This plasmid encoded an endoglucanase (celA) and part of a putative beta-glucosidase (celB), both of which were distinctly different from all previously reported homologues. CelA protein shared limited homology with members of the A3 subfamily of cellulases, being similar to endoglucanase C from Clostridium thermocellum (40% identity). The N-terminal part of CelB protein was most similar to beta-glucosidase from Pseudomonas fluorescens subsp. cellulosa (28% homology). The use of zoolibraries constructed from natural or laboratory enrichment cultures offers the potential to discover many new enzymes for biotechnological applications.

Components of dairy manure management systems.

Dairy manure management systems should account for the fate of excreted nutrients that may be of environmental concern. Currently, regulatory oversight is directed primarily at the assurance of water quality; N is the most monitored element. Land application of manure at acceptable fertilizer levels to crops produced on the farm by hauling or by pumping flushed manure effluent through irrigation systems is the basis of most systems. Nutrient losses to surface and groundwaters can be avoided, and significant economic value can be obtained from manure as fertilizer if adequate crop production is possible. Dairies with insufficient crop production potential need affordable systems to concentrate manure nutrients, thereby reducing hauling costs and possibly producing a salable product. Precipitation of additional nutrients from flushed manures with sedimented solids may be possible. Composting of separated manure solids offers a possible method to stabilize solids for distribution, but, most often, solids separated from dairy manures are fibrous and low in fertility. Manure solids combined with wastes from other sources may have potential if a marketable product can be produced or if sufficient subsidy is received for processing supplementary wastes. Solutions to odor problems are needed. Energy generated from manure organic matter, via anaerobic digestion, reduces atmospheric emissions of methane and odorous compounds. Use of constructed wetlands or harvesting of photosynthetic biomass from wastewater has the potential to improve water quality, making extensive recycling possible.