Soil physicochemical conditions, denitrification rates, and abundance in north Carolina coastal plain restored wetlands.

Over the last century, North Carolina has seen a severe reduction in the percentage of wetlands and a rise in negative environmental impacts related to this loss. To counter these effects, efforts have been enacted to mitigate wetland loss and create new wetland areas. The objective of this study was to assess the impact of hydrological restoration at several sites in the North Carolina coastal plain. Nine sites were selected for study. Hydrologically restored wetlands were compared with natural wetlands and prior converted (PC) croplands (i.e., historic wetlands under agricultural production). Each site was analyzed along a relative wetness gradient, and physicochemical properties, denitrification enzyme activity, and NO reductase gene () abundances using real-time PCR were measured. Physicochemically, restoration resulted in significantly increased levels of total C as compared with PC cropland sites. Restored wetland sites also saw pH, soil moisture, P, and NO+NO approximate levels similar to those of natural wetlands. Denitrification enzyme activity rates varied based on relative wetness within individual sites, generally increasing with increasing soil moisture. However, denitrification tended to be lower in restored wetland sites relative to natural wetlands. Gene abundances of saw statistically significant decreases in restored wetland soils. In conclusion, although analysis of restored wetlands reveals clear changes in several physicochemical characteristics and significant decreases in gene abundances, restoration efforts appear to have not significantly affected the denitrification component of the N cycle.

Denitrification in anaerobic lagoons used to treat swine wastewater.

Anaerobic lagoons are commonly used for the treatment of swine wastewater. Although these lagoons were once thought to be relatively simple, their physical, chemical, and biological processes are very complex. This study of anaerobic lagoons had two objectives: (i) to quantify denitrification enzyme activity (DEA) and (ii) to evaluate the influence of lagoon characteristics on the DEA. The DEA was measured by the acetylene inhibition method. Wastewater samples and physical and chemical measurements were taken from the wastewater column of nine anaerobic swine lagoons from May 2006 to May 2009. These lagoons were typical for anaerobic swine lagoons in the Carolinas relative to their size, operation, and chemical and physical characteristics. Their mean value for DEA was 87 mg N2O-N m(-3) d(-1). In a lagoon with 2-m depth, this rate of DEA would be compatible with 1.74 kg N ha(-1) d(-1) When nonlimiting nitrate was added, the highest DEA was compatible with 4.38 kg N ha(-1) d(-1) loss. Using stepwise regression for this treatment, the lagoon characteristics (i.e., soluble organic carbon, total nitrogen, temperature, and NO3-N) provided a final step model R2 of 0.69. Nitrous oxide from incomplete denitrification was not a significant part of the system nitrogen balance. Although alternate pathways of denitrification may exist within or beneath the wastewater column, this paper documents the lack of sufficient denitrification enzyme activity within the wastewater column of these anaerobic lagoons to support large N2 gas losses via classical nitrification and denitrification.

Designer, acidic biochar influences calcareous soil characteristics.

In a proof-of-concept study, an acidic (pH 5.8) biochar was created using a low pyrolysis temperature (350 °C) and steam activation (800 °C) to potentially improve the soil physicochemical status of an eroded calcareous soil. Biochar was added at 0%, 1%, 2%, and 10% (by wt.) and soils were destructively sampled at 1, 2, 3, 4, 5, and 6 month intervals. Soil was analyzed for gravimetric water content, pH, NO3-N, plant-available Fe, Zn, Mn, Cu, and P, organic C, CO2 respiration, and microbial enumeration via extractable DNA and 16S rRNA gene copies. Gravimetric soil water content increased with biochar application regardless of rate, as compared to the control. Soil pH decreased between 0.2 and 0.4 units, while plant-available Zn, Mn, and P increased with increasing biochar application rate. Micronutrient availability decreased over time likely due to insoluble mineral species precipitation. Increasing biochar application raised the soil organic C content and remained elevated over time. Increasing biochar application rate also increased respired CO2, yet the CO2 released decreased over time. Soil NO3-N concentrations significantly decreased with increasing biochar application rate likely due to microbial immobilization or denitrification. Depending on application rate, biochar produced a 1.4 to 2.1-fold increase in soil DNA extracted and 1.4- to 2.4-fold increase in 16S rRNA gene abundance over control soils, suggesting microbial stimulation and a subsequent burst of activity upon biochar addition. Our results showed that there is promise in designing a biochar to improve the quality and water relations of eroded calcareous soils.

Microbial community structure across a wastewater-impacted riparian buffer zone in the southeastern coastal plain.

Riparian buffer zones are important for both natural and developed ecosystems throughout the world because of their ability to retain nutrients, prevent soil erosion, protect aquatic environments from excessive sedimentation, and filter pollutants. Despite their importance, the microbial community structures of riparian buffer zones remains poorly defined. Our objectives for this study were twofold: first, to characterize the microbial populations found in riparian buffer zone soils; and second, to determine if microbial community structure could be linked to denitrification enzyme activity (DEA). To achieve these objectives, we investigated the microbial populations of a riparian buffer zone located downslope of a pasture irrigated with swine lagoon effluent, utilizing DNA sequencing of the 16S rDNA, DEA, and quantitative PCR (qPCR) of the denitrification genes nirK, nirS, and nosZ. Clone libraries of the 16S rDNA gene were generated from each of twelve sites across the riparian buffer with a total of 986 partial sequences grouped into 654 operational taxonomic units (OTUs). The Proteobacteria were the dominant group (49.8% of all OTUs), with the Acidobacteria also well represented (19.57% of all OTUs). Analysis of qPCR results identified spatial relationships between soil series, site location, and gene abundance, which could be used to infer both incomplete and total DEA rates.

Thermochemical conversion of livestock wastes: carbonization of swine solids.

Slow pyrolysis or carbonization promotes the conversion of animal manures such as swine manure into charcoal. In this paper, the carbonizing kinetics of swine solids taken from different treatment stages were investigated with a thermogravimetric analyzer. Compared to their biologically stabilized counterpart (lagoon sludge) with an activation energy of 160 kJ mol(-1), the activation energies for fresh swine solid samples such as homogenized flushed manure and dewatered solids were much lower between 92 and 95 kJ mol(-1). Compared to the kinetics of first order decomposition of cellulose, the pyrolytic decomposition of the swine manures were more complex with the reaction orders varying at 3.7 and 5.0. The two different mathematical methods employed in this paper yielded the similar values of activation energy (E) and pre-exponential factor (A), confirming the validity of these methods. The results of this study provide useful information for development of farm-scale swine solid carbonization process.

Outer membrane protein B1, an iron-repressible protein conserved in the outer membrane of Moraxella (Branhamella) catarrhalis, binds human transferrin.

Moraxella (Branhamella) catarrhalis is a gram-negative human mucosal pathogen, which primarily causes otitis media in young children. However, this bacterium is also a common cause of lower respiratory tract infections in adults with underlying lung disease. Our previous data have shown that M. catarrhalis expresses iron-repressible outer membrane proteins in response to iron limitation. We have extended these observations to demonstrate that one of these proteins, termed outer membrane protein (OMP) B1, binds human transferrin. Using a newly developed monoclonal antibody to OMP B1, we determined that this protein is conserved in the iron-stressed outer membranes of all clinical isolates of M. catarrhalis tested to date. Furthermore, our data have confirmed that children infected with M. catarrhalis have immunoglobulin G antibodies to OMP B1 in their convalescent sera. These current data suggest that OMP B1 is immunogenic and expressed in vivo and may be involved in an iron uptake mechanism utilized by M. catarrhalis.

Reduced virulence of a Bordetella bronchiseptica siderophore mutant in neonatal swine.

One means by which Bordetella bronchiseptica scavenges iron is through production of the siderophore alcaligin. A nonrevertible alcaligin mutant derived from the virulent strain 4609, designated DBB25, was constructed by insertion of a kanamycin resistance gene into alcA, one of the genes essential for alcaligin biosynthesis. The virulence of the alcA mutant in colostrum-deprived, caesarean-delivered piglets was compared with that of the parent strain in two experiments. At 1 week of age, piglets were inoculated with phosphate-buffered saline, 4609, or DBB25. Two piglets in each group were euthanatized on day 10 postinfection. The remainder were euthanatized at 21 days postinfection. Clinical signs, including fever, coughing, and sneezing, were present in both groups. Nasal washes performed 7, 14, and 21 days postinoculation demonstrated that strain DBB25 colonized the nasal cavity but did so at levels that were significantly less than those achieved by strain 4609. Analysis of colonization based on the number of CFU per gram of tissue recovered from the turbinate, trachea, and lung also demonstrated significant differences between DBB25 and 4609, at both day 10 and day 21 postinfection. Mild to moderate turbinate atrophy was apparent in pigs inoculated with strain 4609, while turbinates of those infected with strain DBB25 developed no or mild atrophy. We conclude from these results that siderophore production by B. bronchiseptica is not essential for colonization of swine but is required for maximal virulence. B. bronchiseptica mutants with nonrevertible defects in genes required for alcaligin synthesis may be candidates for evaluation as attenuated, live vaccine strains in conventionally reared pigs.