Manure-fertilized Soil Research Articles

The impacts of different long-term fertilization regimes on the bioavailability of arsenic in soil: integrating chemical approach with Escherichia coli arsRp::luc-based biosensor

An Escherichia coli arsRp::luc-based biosensor was constructed to measure the bioavailability of arsenic (As) in soil. In previous induction experiments, it produced a linear response (R (2) = 0.96, P < 0.01) to As from 0.05 to 5μmol/L after a 2-h incubation. Then, both chemical sequential extraction, Community Bureau of Reference recommended sequential extraction procedures (BCR-SEPs) and E. coli biosensor, were employed to assess the impact of different long-term fertilization regimes containing N, NP, NPK, M (manure), and NPK + M treatments on the bioavailability of arsenic (As) in soil. Per the BCR-SEPs analysis, the application of M and M + NPK led to a significant (P < 0.01) increase of exchangeable As (2-7 times and 2-5 times, respectively) and reducible As (1.5-2.5 times and 1.5-2.3 times, respectively) compared with the no fertilization treated soil (CK). In addition, direct contact assay of E. coli biosensor with soil particles also supported that bioavailable As in manure-fertilized (M and M + NPK) soil was significantly higher (P < 0.01) than that in CK soil (7 and 9 times, respectively). Organic carbon may be the major factor governing the increase of bioavailable As. More significantly, E. coli biosensor-determined As was only 18.46-85.17% of exchangeable As and 20.68-90.1% of reducible As based on BCR-SEPs. In conclusion, NKP fertilization was recommended as a more suitable regime in As-polluted soil especially with high As concentration, and this E. coli arsRp::luc-based biosensor was a more realistic approach in assessing the bioavailability of As in soil since it would not overrate the risk of As to the environment.

Carbon Mineralization and Microbial Attributes in Straw-Amended Soils as Affected by Moisture Levels

An 80-d incubation experiment was conducted to investigate straw decomposition, the priming effect and microbial characteristics in a non-fertilized soil (soil 1) and a long-term organic manure-fertilized soil (soil 2) with and without 13C-labeled maize straw amendment under different moisture levels. The soil 2 showed a markedly higher priming effect, microbial biomass C (Cmic), and β-glucosidase activity, and more abundant populations of bacteria and fungi than the soil 1. Also, soil CO2 emission, Cmic, β-glucosidase activity, and bacterial and fungal population sizes were substantially enhanced by straw amendment. In the presence of straw, the amount of straw mineralization and assimilation by microbes in the soil at 55% of water holding capacity (WHC) were significantly higher by 31% and 17%, respectively, compared to those at 25% of WHC. In contrast, β-glucosidase activity and fungal population size were both enhanced as the moisture content decreased. Cmic decreased as straw availability decreased, which was mainly attributed to the reduction of straw-derived Cmic. Amended soils, except the amended soil 2 at 25% of WHC, had a more abundant fungal population as straw availability decreased, indicating that fungal decomposability of added straw was independent of straw availability. Non-metric multidimensional scaling analysis based on fungal denatured gradient gel electrophoresis band patterns showed that shifts in the fungal community structure occurred as water and straw availability varied. The results indirectly suggest that soil fungi are able to adjust their degradation activity to water and straw availability by regulating their community structure.

Impact of Manure Fertilization on the Abundance of Antibiotic-Resistant Bacteria and Frequency of Detection of Antibiotic Resistance Genes in Soil and on Vegetables at Harvest

Consumption of vegetables represents a route of direct human exposure to bacteria found in soil. The present study evaluated the complement of bacteria resistant to various antibiotics on vegetables often eaten raw (tomato, cucumber, pepper, carrot, radish, lettuce) and how this might vary with growth in soil fertilized inorganically or with dairy or swine manure. Vegetables were sown into field plots immediately following fertilization and harvested when of marketable quality. Vegetable and soil samples were evaluated for viable antibiotic-resistant bacteria by plate count on Chromocult medium supplemented with antibiotics at clinical breakpoint concentrations. DNA was extracted from soil and vegetables and evaluated by PCR for the presence of 46 gene targets associated with plasmid incompatibility groups, integrons, or antibiotic resistance genes. Soil receiving manure was enriched in antibiotic-resistant bacteria and various antibiotic resistance determinants. There was no coherent corresponding increase in the abundance of antibiotic-resistant bacteria enumerated from any vegetable grown in manure-fertilized soil. Numerous antibiotic resistance determinants were detected in DNA extracted from vegetables grown in unmanured soil. A smaller number of determinants were additionally detected on vegetables grown only in manured and not in unmanured soil. Overall, consumption of raw vegetables represents a route of human exposure to antibiotic-resistant bacteria and resistance determinants naturally present in soil. However, the detection of some determinants on vegetables grown only in freshly manured soil reinforces the advisability of pretreating manure through composting or other stabilization processes or mandating offset times between manuring and harvesting vegetables for human consumption.

Impact of 12-year field treatments with organic and inorganic fertilizers on crop productivity and mycorrhizal community structure

The effect of manure and mineral fertilization on the arbuscular mycorrhizal (AM) fungal community structure of sunflower (Helianthus annuus L.) plants was studied. Soils were collected from a field experiment treated for 12 years with equivalent nitrogen (N) doses of inorganic N, dairy manure slurry, or without N fertilization. Fresh roots of tall fescue (Festuca arundinacea Schreb.) grass collected from the field plots without N fertilization and unfumigated field soils were used as native microbial inoculum sources. Sunflower plants were sown in pots containing these soils, and three different means of manipulating the microbial community were set: unfumigated soil with fresh grass roots, fumigated soil with fresh grass roots, or fumigated soil with sterilized grass roots. Assessing the implications with respect to plant productivity and mycorrhizal community structure was investigated. Twelve AM fungal OTUs were identified from root or soil samples as different taxa of Acaulospora, Claroideoglomus, Funneliformis, Rhizophagus, and uncultured Glomus, using PCR-DGGE and sequencing of an 18S rRNA gene fragment. Sunflower plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral N or unfertilized, with an abundance of Rhizophagus intraradices-like (B2). The results also showed that AM inoculation increased P and N contents in inorganic N-fertilized or unfertilized plants, but not in manure-fertilized plants.

Response of Induced Perturbation on Replicating β-Proteobacterial Ammonia-Oxidizing Populations in Soil

The short-term response of induced perturbation by 4-ethylphenol on β-proteobacterial ammonia oxidizers (β-AOB) was investigated in two soils with initial differences in community structure. The hypotheses were that short-term effects of a disturbance of the AOB community is best monitored by specifically looking at the active populations and that soils with dissimilar active AOB populations would display different degree of resistance or resilience. Two soils from a previously characterized long-term field study fertilized with manure or sewage sludge was used. Soil microcosms were incubated in the laboratory over 15 days. The substrate-induced ammonia oxidation was measured, and the composition of β-AOB communities was determined by PCR-DGGE of specific β-AOB 16S rRNA gene fragments. Actively replicating members of the β-AOB were distinguished by the use of bromodeoxyuridine (BrdU) immunocapture. This approach demonstrated that only a minor fraction of the total AOB community was active. Exposure to 4-ethylphenol resulted in approximately 90% lowered substrate-induced ammonia oxidation rates in both soils. This activity inhibition was not accompanied by shifts in β-AOB community structure when total β-AOB DNA was studied. By contrast, changes were seen in the DGGE banding pattern of the BrdU-labeled community DNA after 4-ethylphenol addition in the manure-fertilized soil. In the sewage sludge fertilized soil, the banding pattern of the BrdU-labeled β-AOB remained unchanged, but bands were weaker after the disturbance. In conclusion, it was shown that BrdU immunocapture was applicable to detect shifts in community composition among replicating β-AOB populations in soil. However, this was not reflected by the soils' ammonia oxidation capacity to resist to or recover from the induced perturbation suggesting that rapid population shifts may not influence soil functioning in a short-term perspective.

Population size and specific nitrification potential of soil ammonia-oxidizing bacteria under long-term fertilizer management

Population size of soil ammonia-oxidizing bacteria (AOB) was quantified by real-time PCR in a long-term (16 years) field experiment under different fertilizer managements. AOB population sizes in mineral nitrogen-fertilized soils and organic manure-fertilized soil were 10.3 and 3.1 times, respectively, that of the control, while phosphorus and potassium fertilization had no significant effect. On the other hand, the AOB specific nitrification potential (soil nitrification potential per AOB cell) was significantly higher ( P < 0.05) in organic manure-fertilized soil than in mineral-fertilized soils and the control, indicating that AOB was likely more metabolically active in organic manure-fertilized soils than in mineral nitrogen-fertilized soils after long-term application.

Soil microbial biomass, dehydrogenase activity, bacterial community structure in response to long-term fertilizer management

This study describes the effects of balanced versus nutrient-deficiency fertilization on soil microbial biomass, activity, and bacterial community structure in a long-term (16 years) field experiment. Long-term fertilization greatly increased soil microbial biomass C and dehydrogenase activity, except that the P-deficiency fertilization had no significant effect. Organic manure had a significantly greater ( P<0.05) impact on the biomass C and the activity, compared with mineral fertilizers. Microbial metabolic activity (dehydrogenase activity per microbial biomass C) was significantly higher ( P<0.05) under balanced fertilization than under nutrient-deficiency fertilization. General bacterial community structure was analyzed by PCR-denaturing gradient gel electrophoresis (DGGE) targeting eubacterial 16S rRNA gene. Mineral fertilization did not affect the DGGE banding pattern, while specific DGGE band was observed in organic manure-fertilized soils. Phylogenetic analysis showed that the change of bacterial community in organic manure-fertilized soil might not be because of the direct influence of the bacteria in the compost, but because of the promoting effect of the compost on the growth of an indigenous Bacillus sp. in the soil. We emphasize the importance of balanced-fertilization, as well as the role of P, in maintaining soil organic matter, and promoting the biomass and activity of microorganisms.

Evaluation of Fertilization-to-Planting and Fertilization-to-Harvest Intervals for Safe Use of Noncomposted Bovine Manure in Wisconsin Vegetable Production

Fresh bovine manure was mechanically incorporated into loamy sand and silty clay loam Wisconsin soils in April 2004. At varying fertilization-to-planting intervals, radish, lettuce, and carrot seeds were planted; crops were harvested 90, 100, 110 or 111, and 120 days after manure application. As an indicator of potential contamination with fecal pathogens, levels of Escherichia coli in the manure-fertilized soil and presence of E. coli on harvested vegetables were monitored. From initial levels of 4.0 to 4.2 log CFU/g, E. coli levels in both manure-fertilized soils decreased by 2.4 to 2.5 log CFU/g during the first 7 weeks. However, E. coli was consistently detected from enriched soil samples through week 17, perhaps as a result of contamination by birds and other wildlife. In the higher clay silty clay loam soil, the fertilization-to-planting interval affected the prevalence of E. coli on lettuce but not on radishes and carrots. Root crop contamination was consistent across different fertilization-to-harvest intervals in silty clay loam, including the National Organic Program minimum fertilization-to-harvest interval of 120 days. However, lettuce contamination in silty clay loam was significantly (P < 0.10) affected by fertilization-to-harvest interval. Increasing the fertilization-to-planting interval in the lower clay loamy sand soil decreased the prevalence of E. coli on root crops. The fertilization-to-harvest interval had no clear effect on vegetable contamination in loamy sand. Overall, these results do not provide grounds for reducing the National Organic Program minimum fertilization-to-harvest interval from the current 120-day standard.

Escherichia coli contamination of vegetables grown in soils fertilized with noncomposted bovine manure: garden-scale studies.

In this study we tested the validity of the National Organic Program (NOP) requirement for a > or =120-day interval between application of noncomposted manure and harvesting of vegetables grown in manure-fertilized soil. Noncomposted bovine manure was applied to 9.3-m2 plots at three Wisconsin sites (loamy sand, silt loam, and silty clay loam) prior to spring and summer planting of carrots, radishes, and lettuce. Soil and washed (30 s under running tap water) vegetables were analyzed for indigenous Escherichia coli. Within 90 days, the level of E. coli in manure-fertilized soil generally decreased by about 3 log CFU/g from initial levels of 4.2 to 4.4 log CFU/g. Low levels of E. coli generally persisted in manure-fertilized soil for more than 100 days and were detected in enriched soil from all three sites 132 to 168 days after manure application. For carrots and lettuce, at least one enrichment-negative sample was obtained < or =100 days after manure application for 63 and 88% of the treatments, respectively. The current > or =120-day limit provided an even greater likelihood of not detecting E. coli on carrots (> or =1 enrichment-negative result for 100% of the treatments). The rapid maturation of radishes prevented conclusive evaluation of a 100- or 120-day application-to-harvest interval. The absolute absence of E. coli from vegetables harvested from manure-fertilized Wisconsin soils may not be ensured solely by adherence to the NOP > or =120-day limit. Unless pathogens are far better at colonizing vegetables than indigenous E. coli strains are, it appears that the risk of contamination for vegetables grown in Wisconsin soils would be elevated only slightly by reducing the NOP requirement to > or =100 days.

Salmonella enterica serovar Typhimurium and Escherichia coli contamination of root and leaf vegetables grown in soils with incorporated bovine manure.

Bovine manure, with or without added Salmonella enterica serovar Typhimurium (three strains), was incorporated into silty clay loam (SCL) and loamy sand (LS) soil beds (53- by 114-cm surface area, 17.5 cm deep) and maintained in two controlled-environment chambers. The S. enterica serovar Typhimurium inoculum was 4 to 5 log CFU/g in manure-fertilized soil. The conditions in the two environmental chambers, each containing inoculated and uninoculated beds of manure-fertilized soil, simulated daily average Madison, Wis., weather conditions (hourly temperatures, rainfall, daylight, and humidity) for a 1 March or a 1 June manure application and subsequent vegetable growing seasons ending 9 August or 28 September, respectively. Core soil samples were taken biweekly from both inoculated and uninoculated soil beds in each chamber. Radishes, arugula, and carrots were planted in soil beds, thinned, and harvested. Soils, thinned vegetables, and harvested vegetables were analyzed for S. enterica serovar Typhimurium and Escherichia coli (indigenous in manure). After the 1 March manure application, S. enterica serovar Typhimurium was detected at low levels in both soils on 31 May, but not on vegetables planted 1 May and harvested 12 July from either soil. After the 1 June manure application, S. enterica serovar Typhimurium was detected in SCL soil on 7 September and on radishes and arugula planted in SCL soil on 15 August and harvested on 27 September. In LS soil, S. enterica serovar Typhimurium died at a similar rate (P >or= 0.05) after the 1 June manure application and was less often detected on arugula and radishes harvested from this soil compared to the SCL soil. Pathogen levels on vegetables were decreased by washing. Manure application in cool (daily average maximum temperature of <10 degrees C) spring conditions is recommended to ensure that harvested vegetables are not contaminated with S. enterica serovar Typhimurium. Manure application under warmer (daily average maximum temperature >20 degrees C) summer conditions is not recommended when vegetable planting is done between the time of manure application and late summer. A late fall manure application will not increase the risk of contaminating vegetables planted the next spring, since further experiments showed that repeated freeze-thaw cycles were detrimental to the survival of S. enterica serovar Typhimurium and E. coli in manure-fertilized soil. The number of indigenous E. coli in soil was never significantly lower (P < 0.05) than that of S. enterica serovar Typhimurium, suggesting its usefulness as an indicator organism for evaluating the risk of vegetable contamination with manure-borne S. enterica serovar Typhimurium.