Effects of Subsurface Banding and Broadcast of Poultry Litter and Cover Crop on Soil Microbial Populations.

Agronomic management is aimed at managing the crop environment to maximize crop yield, but soil biology is often ignored. This study aimed to compare the application of poultry litter via broadcast and subsurface banding versus standard inorganic fertilizer to cotton ( L.) and their effects on soil bacterial populations and fecal indicator bacteria. The study comprised a randomized complete block design, with fertilizer and time of application as treatment effects and cover crop as a main effect. Soil cores were collected and analyzed from 2008 to 2014. Fecal indicator bacteria were at detection limits for all treatments, where the integron 1 gene was significantly elevated in litter plots. There were few differences between litter application approaches, but both significantly increased key biogeochemical genes over control plots, whereas a cover crop only increased soil moisture and urease C. Data suggested a positive residual effect of litter application with 16S, phosphatase A, and urease C genes elevated over controls, but similar to standard fertilizer plots. High-throughput 16S ribosomal RNA analysis suggested increased diversity and enrichment indices in litter and standard fertilizer over untreated control plots. Litter and standard fertilizer effects persisted 4 and 2 yr after application, respectively, as evidenced by residual library community structures. This study demonstrated the positive effects of litter application on the soil bacterial community when compared with untreated control plots. Some differences between standard fertilization and litter practices were noted and suggest that there is a positive residual effect on soil microbial populations associated with both practices.

Broiler Litter × Industrial By-Products Reduce Nutrients and Microbial Losses in Surface Runoff When Applied to Forages.

The inability to incorporate broiler litter (BL) into permanent hayfields and pastures leads to nutrient accumulation near the soil surface and increases the potential transport of nutrients in runoff. This study was conducted on Marietta silt loam soil to determine the effect of flue gas desulfurization (FGD) gypsum and lignite on P, N, C, and microbial concentrations in runoff. Treatments were (i) control (unfertilized) and (ii) BL at 13.4 Mg ha alone or (iii) treated with either FGD gypsum or lignite applied at 20% (w/w) (2.68 Mg ha). Rainfall simulators were used to produce a 5.6 cm h storm event sufficient in duration to cause 15 min of continuous runoff. Repeated rains were applied at 3-d intervals to determine how long FGD gypsum and lignite are effective in reducing loss of litter-derived N, P, and C from soil. Application of BL increased N, P, and C concentrations in runoff as compared to the control. Addition of FGD gypsum reduced ( < 0.05) water-soluble P and dissolved organic C concentrations in runoff by 39 and 16%, respectively, as compared to BL alone. Lignite reduced runoff total N and NH-N concentrations by 38 and 70%, respectively, as compared to BL alone. Addition of FGD gypsum or lignite failed to significantly reduce microbial loads in runoff, although both treatments reduced microbial concentration by >20%. Thus, BL treated with FGD and lignite can be considered as cost-effective management practices in the mitigation of P, N, and C and possibly microbial concentration in runoff.

Composting and gypsum amendment of broiler litter to reduce nutrient leaching loss.

The effect of composted litter relative to fresh litter on leaching losses of nutrients has not been well documented. Fresh and composted broiler litter was surface-applied to bermudagrass (hay) [ (L.) Pers.] established in undisturbed soil columns based on N need of the grass in the presence or absence of flue gas desulfurization (FGD) gypsum to evaluate an approach to reduce broiler litter nutrient leaching potential. Columns were periodically leached and biomass was harvested during the 60-d experiment. Total N applied to bermudagrass from broiler litter was 320 kg ha. Gypsum was mixed with fresh and composted litter at the rate based on 20% of litter weight. For composted broiler litter, NO-N, P, K, Cu, and Zn contents in the leachate obtained from the first leaching event were 58, 50, 40, 32, and 38% less than fresh broiler litter, respectively. Significant decreases in NO-N (13%), P (53%), Cu (17%), and Zn (28%) in leachate were obtained when gypsum was mixed with fresh broiler litter. Fresh broiler litter and composted broiler litter applications increased bermudagrass growth compared with the control and gypsum significantly increased yields when mixed with broiler litter. Composted broiler litter application significantly increased N and organic C in the soil compared with fresh litter. Results demonstrate that coapplication of composted broiler litter with FGD gypsum provide the most effective management option for minimizing leaching losses of nutrients while sustaining crop productivity.

Effects of bedding materials in applied poultry litter and immobilizing agents on runoff water, soil properties, and bermudagrass growth.

Poultry producers in the United States have begun using different types of bedding materials in production houses. Release into the environment of nutrients from applied poultry litter (PL) made with different bedding materials has not been investigated, and little information is available on nutrient concentrations in soils that receive broiler litter made with such materials. In this greenhouse study, two bedding materials (rice hulls and pine chips) in PL and two nutrient-immobilizing agents (gypsum and biochar) were applied to bermudagrass, and chemical and microbial contents of runoff water, soil properties, and plant growth were evaluated. Treatments with rice hull bedding material in PL had less runoff nutrient and greater soil soluble N and P compared with pine chip bedding. Gypsum and biochar both significantly reduced C, N, P, Cu, and Zn losses from the first runoff event, which were reduced by 26, 30, 37, 38, and 38% and by 25, 24, 30, 29, and 35%, respectively, but only gypsum obviously reduced these nutrients from later events. Potassium, Ca, Mg, and Mn increased by 2, 36, 11, and 9 times, respectively, and soluble P, Cu, and Fe significantly decreased by 68, 72, and 98%, respectively, in soil amended with gypsum. Rice hull PL in combination with gypsum significantly increased the growth of bermudagrass. Our results indicate that rice hull PL posed less risk for nutrient loss than pine chip PL when applied to fields and that gypsum was better than biochar for reducing runoff C, N, P, and Cu.

Phenological stage and vegetation index for predicting corn yield under rainfed environments.

Uncrewed aerial systems (UASs) provide high temporal and spatial resolution information for crop health monitoring and informed management decisions to improve yields. However, traditional in-season yield prediction methodologies are often inconsistent and inaccurate due to variations in soil types and environmental factors. This study aimed to identify the best phenological stage and vegetation index (VI) for estimating corn yield under rainfed conditions. Multispectral images were collected over three years (2020-2022) during the corn growing season and over fifty VIs were analyzed. In the three-year period, thirty-one VIs exhibited significant correlations (r ≥ 0.7) with yield. Sixteen VIs were significantly correlated with the yield at least for two years, and five VIs had a significant correlation with the yield for all three years. A strong correlation with yield was achieved by combining red, red edge, and near infrared-based indices. Further, combined correlation and random forest an alyses between yield and VIs led to the identification of consistent and highest predictive power VIs for corn yield prediction. Among them, leaf chlorophyll index, Medium Resolution Imaging Spectrometer (MERIS) terrestrial chlorophyll index and modified normalized difference at 705 were the most consistent predictors of corn yield when recorded around the reproductive stage (R1). This study demonstrated the dynamic nature of canopy reflectance and the importance of considering growth stages, and environmental conditions for accurate corn yield prediction.

Plant-based FRET biosensor discriminates environmental zinc levels.

Heavy metal accumulation in the environment poses great risks to flora and fauna. However, monitoring sites prone to accumulation poses scale and economic challenges. In this study, we present and test a method for monitoring these sites using fluorescent resonance energy transfer (FRET) change in response to zinc (Zn) accumulation in plants as a proxy for environmental health. We modified a plant Zn transport protein by adding flanking fluorescent proteins (FPs) and deploying the construct into two different species. In Arabidopsis thaliana, FRET was monitored by a confocal microscope and had a 1.4-fold increase in intensity as the metal concentration increased. This led to a 16.7% overall error-rate when discriminating between a control (1µm Zn) and high (10mm Zn) treatment after 96h. The second host plant (Populus tremula×Populu salba) also had greater FRET values (1.3-fold increase) when exposed to the higher concentration of Zn, while overall error-rates were greater at 22.4%. These results indicate that as plants accumulate Zn, protein conformational changes occur in response to Zn causing differing interaction between FPs. This results in greater FRET values when exposed to greater amounts of Zn and monitored with appropriate light sources and filters. We also demonstrate how this construct can be moved into different host plants effectively including one tree species. This chimeric protein potentially offers a method for monitoring large areas of land for Zn accumulation, is transferable among species, and could be modified to monitor other specific heavy metals that pose environmental risks.

Poplar maintains zinc homeostasis with heavy metal genes HMA4 and PCS1.

Perennial woody species, such as poplar (Populus spp.) must acquire necessary heavy metals like zinc (Zn) while avoiding potential toxicity. Poplar contains genes with sequence homology to genes HMA4 and PCS1 from other species which are involved in heavy metal regulation. While basic genomic conservation exists, poplar does not have a hyperaccumulating phenotype. Poplar has a common indicator phenotype in which heavy metal accumulation is proportional to environmental concentrations but excesses are prevented. Phenotype is partly affected by regulation of HMA4 and PCS1 transcriptional abundance. Wild-type poplar down-regulates several transcripts in its Zn-interacting pathway at high Zn levels. Also, overexpressed PtHMA4 and PtPCS1 genes result in varying Zn phenotypes in poplar; specifically, there is a doubling of Zn accumulation in leaf tissues in an overexpressed PtPCS1 line. The genomic complement and regulation of poplar highlighted in this study supports a role of HMA4 and PCS1 in Zn regulation dictating its phenotype. These genes can be altered in poplar to change its interaction with Zn. However, other poplar genes in the surrounding pathway may maintain the phenotype by inhibiting drastic changes in heavy metal accumulation with a single gene transformation.

Nutrients and bacteria in common contiguous Mississippi soils with and without broiler litter fertilization.

In Mississippi, spent poultry litter is used as fertilizer. Nutrient and bacterial levels in litter and nutrient levels in litter-fertilized (L+) soil are known, but less is known of bacterial levels in L+ soil. This study compared contiguous L+ and non-litter-fertilized (L-) soils comprising 15 soil types on five farms in April through May 2009. Levels of pH; NO-N; and Mehlich-3-extractable (M3) and water-extractable (WE) P, Ca, K, and Cu were higher in L+ than in L- soil. Total C; total N; NH-N; and M3 and WE Na, Fe, and Zn did not differ in L+ and L- soil. Bacterial levels were higher in 0- to 5-cm than in 5- to 10-cm cores. Levels were higher in L+ than in L- soil for culturally determined heterotrophic plate counts and staphylococci and were lower for total bacteria estimated by quantitative polymerase chain reaction (qPCR) of 16S rRNA, but cultural levels of thermotolerant coliforms, , , and enterococci were not different. Cultural presence/absence (CPA) tests and qPCR for spp., spp., and spp. detected only spp., which did not differ in L+ (CPA = 77% positive samples; mean qPCR = 0.65 log genomic units [gu] g) and L- (CPA = 70% positive samples; mean qPCR = 0 log gu g) soils. Litter applications were associated with higher levels of pH, P, Cu, heterotrophic plate counts, and staphylococci. Fecal indicator and enteric pathogen levels were not affected. We conclude that, although some litter-derived nutrients and bacteria persisted between growing seasons in L+ soils, enteric pathogens did not.

Apoplast proteome reveals that extracellular matrix contributes to multistress response in poplar.

BACKGROUND: Riverine ecosystems, highly sensitive to climate change and human activities, are characterized by rapid environmental change to fluctuating water levels and siltation, causing stress on their biological components. We have little understanding of mechanisms by which riverine plant species have developed adaptive strategies to cope with stress in dynamic environments while maintaining growth and development. RESULTS: We report that poplar (Populus spp.) has evolved a systems level "stress proteome" in the leaf-stem-root apoplast continuum to counter biotic and abiotic factors. To obtain apoplast proteins from P. deltoides, we developed pressure-chamber and water-displacement methods for leaves and stems, respectively. Analyses of 303 proteins and corresponding transcripts coupled with controlled experiments and bioinformatics demonstrate that poplar depends on constitutive and inducible factors to deal with water, pathogen, and oxidative stress. However, each apoplast possessed a unique set of proteins, indicating that response to stress is partly compartmentalized. Apoplast proteins that are involved in glycolysis, fermentation, and catabolism of sucrose and starch appear to enable poplar to grow normally under water stress. Pathogenesis-related proteins mediating water and pathogen stress in apoplast were particularly abundant and effective in suppressing growth of the most prevalent poplar pathogen Melampsora. Unexpectedly, we found diverse peroxidases that appear to be involved in stress-induced cell wall modification in apoplast, particularly during the growing season. Poplar developed a robust antioxidative system to buffer oxidation in stem apoplast. CONCLUSION: These findings suggest that multistress response in the apoplast constitutes an important adaptive trait for poplar to inhabit dynamic environments and is also a potential mechanism in other riverine plant species.

Comparison of selected nutrients and bacteria from common contiguous soils inside and outside swine lagoon effluent spray fields after long-term use.

Swine (Sus scofa domestica) lagoon effluent is a valuable resource. In the U.S. Mid-South it is applied from April to September to fertilize grass hay in spray-irrigated fields. Lagoon levels of nutrients and bacteria, and soil levels of nutrients have been documented, but little was known of effluent bacterial levels in soil. The present study examined levels of selected effluent bacteria and nutrients in soils inside and outside spray fields after >15 yr of effluent irrigation. Samples were collected February to March 2009 from contiguous soils spanning adjacent irrigated and nonirrigated areas. Separate soil cores for bacterial and nutrient tests were collected in pairs <10 cm apart. Five cores each were collected at 15-m intervals and combined, respectively, to comprise inside and outside samples from each of 20 soils (four each from five farms/spray fields). Analyses of data combined across all soils showed higher pH and Mehlich-3-extracrable (M3-) P, Mg, K, Na, Cu, and Zn inside than outside spray fields, while total N, total C, M3-Ca, and M3-Mn did not differ. Bacterial levels were higher inside than outside spray fields for heterotrophic plate counts, thermotolerant coliforms, Staphylococcus spp., and Clostridium perfringens, but levels of Escherichia coli and Enterococcus spp. were not different. Cultural presence/absence tests for three pathogens (Listeria spp., Campylobacter spp., and Salmonella spp.) detected only Listeria spp., which did not differ inside (23% positive samples) and outside (28% positive). Molecular tests detected all three pathogens at low levels that were not different inside and outside. We found no evidence of cumulative buildup of Campylobacter spp., Listeria spp., or Salmonela s. in spray field soils.

Characterization of selected nutrients and bacteria from anaerobic swine manure lagoons on sow, nursery, and finisher farms in the Mid-South USA.

Swine (Sus scrofa domestica) production in the Mid-South USA comprises sow, nursery, and finisher farms. A 2007 packing plant closure started a regional shift from finisher to sow and nursery farms. Changes in manure stored in lagoons and land-applied as fertilizer were expected but were unknown because nutrient and bacterial levels had not been characterized by farm type. The objectives of this study were to quantify selected nutrients and bacteria, compare levels by farm types, and project impacts of production shifts. Nutrients and bacteria were characterized in 17 sow, 10 nursery, and 10 finisher farm lagoons. Total and thermotolerant coliforms, Escherichia coli (Migula) Castellani and Chalmers, Enterococcus spp., Clostridium perfringens (Veillon and Zuber) Hauduroy et al., Campylobacter spp., Listeria spp., and Salmonella spp. were evaluated. Highest levels were from total coliforms (1.4- 5.7x10(5) cfu 100 mL(-1)), which occurred with E. coli, Campylobacter spp., C. perfringens, and Enterococcus spp., in every lagoon and virtually every sample. Lowest levels were from Listeria spp. and Salmonella spp. (

Rainfall simulation in greenhouse microcosms to assess bacterial-associated runoff from land-applied poultry litter.

Runoff water following a rain event is one possible source of environmental contamination after a manure application. This greenhouse study used a rainfall simulator to determine bacterial-associated runoff from troughs of common bermudagrass [Cynodon dactylon (L.) Pers.] that were treated with P-based, N-based, and N plus lime rates of poultry (Gallus gallus) litter, recommended inorganic fertilizer, and control. Total heterotrophic plate count (HPC) bacteria, total and thermotolerant coliforms, enterococci, staphylococci, Clostridium perfringens, Salmonella, and Campylobacter, as well as antibiotic resistance profiles for the staphylococci and enterococci isolates were all monitored in runoff waters. Analysis following five rainfall events indicated that staphylococci, enterococci, and clostridia levels were related to manure application rate. Runoff release of staphylococci, enterococci, and C. perfringens were approximately 3 to 6 log10 greater in litter vs. control treatment. In addition, traditional indicators such as thermotolerant and total coliforms performed poorly as fecal indicators. Some isolated enterococci demonstrated increased antibiotic resistance to polymixin b and/or select aminoglyocosides, while many staphylococci were susceptible to most antimicrobials tested. Results indicated poultry litter application can lead to microbial runoff following simulated rain events. Future studies should focus on the use of staphylococci, enterococci, and C. perfringens as indicators.

Bacterial Community Structure Recovery in Reclaimed Coal Mined Soil under Two Vegetative Regimes.

Coal mining can be deleterious to the soil physical and chemical makeup, but also to the soil microbial community. Effectively, the removal of nearly all organic matter from the upper soil horizons reduces the effectiveness of any soil to support vegetation, and up until recently, microbial community parameters were not considered in the successful reclamation of overburden. Thus, our study proposes to measure the uncultivated bacterial community using 16S ribosomal RNA (rRNA) high-throughput sequencing in a chronosequence of reclaimed overburden in Mississippi. The study sites comprised samplings of pasture and wooded reclamation sites consisting of 1 to 13 yr post reclamation time, as well as reference sites. Overall, the primary driver of bacterial community dynamics was vegetative cover, although time also influenced dynamics. Richness estimations for operational taxonomic units (OTUs) showed that recently reclaimed (∼1 yr) and Pasture sites were more OTU rich with levels of >1400 compared with reference site levels of ∼1000. Diversity levels also followed a similar trend. Community structure typically differed between time points and vegetative cover; however, membership was similar between sites and reference, indicating that new communities still shared some membership from the previous community. Overall, physicochemical properties trended toward more positive for soil health as time progressed, but bacterial community recovery was still not structurally recovered, although richness and diversity values exceeded reference. Overall, this study demonstrated that mine reclamation using pasture and/or wood restoration can reestablish the bacterial community to approximate reference conditions, but vegetation is still the dominating environmental factor dictating microbial community.

Influence of land use and land cover on the spatial variability of dissolved organic matter in multiple aquatic environments.

Water quality of lakes, estuaries, and coastal areas serves as an indicator of the overall health of aquatic ecosystems as well as the health of the terrestrial ecosystem that drains to the water body. Land use and land cover plays not only a significant role in controlling the quantity of the exported dissolved organic matter (DOM) but also influences the quality of DOM via various biogeochemical and biodegradation processes. We examined the characteristics and spatial distribution of DOM in five major lakes, in an estuary, and in the coastal waters of the Mississippi, USA, and investigated the influence of the land use and land cover of their watersheds on the DOM composition. We employed absorption and fluorescence spectroscopy including excitation-emission matrix (EEM) combined with parallel factor (PARAFAC) analysis modeling techniques to determine optical properties of DOM and its characteristics in this study. We developed a site-specific PARAFAC model to evaluate DOM characteristics resulting in five diverse DOM compositions that included two terrestrial humic-like (C1 and C3), two microbial humic-like (C2 and C5), and one protein-like (C4) DOM. Our results showed elevated fluorescence levels of microbial humic-like or protein-like DOM in the lakes and coastal waters, while the estuarine waters showed relatively high fluorescence levels of terrestrial humic-like DOM. The results also showed that percent forest and wetland coverage explained 68 and 82% variability, respectively, in terrestrial humic-like DOM exports, while 87% variability in microbially derived humiclike DOM was explained by percent agricultural lands. Strong correlations between microbial humic-like DOM and fluorescence-derived DOM indices such as biological index (BIX) and fluorescence index (FI) indicated autochthonous characteristics in the lakes, while the estuary showed largely allochthonous DOM of terrestrial origin. We also observed higher concentrations of total dissolved phosphorous (TDP) and ammonium nitrogen (NH4-N) in coastal waters potentially due to photodegradation of refractory DOM derived from the sediment-bound organic matter in the coastal wetlands. This study highlights the relationships between the DOM compositions in the water and the land use and land cover in the watershed. The spatial variability of DOM in three different types of aquatic environments enhances the understanding of the role of land use and land cover in carbon cycling through export of organic matter to the aquatic ecosystems..

Microbial ecology, bacterial pathogens, and antibiotic resistant genes in swine manure wastewater as influenced by three swine management systems.

The environmental influence of farm management in concentrated animal feeding operations (CAFO) can yield vast changes to the microbial biota and ecological structure of both the pig and waste manure lagoon wastewater. While some of these changes may not be negative, it is possible that CAFOs can enrich antibiotic resistant bacteria or pathogens based on farm type, thereby influencing the impact imparted by the land application of its respective wastewater. The purpose of this study was to measure the microbial constituents of swine-sow, -nursery, and -finisher farm manure lagoon wastewater and determine the changes induced by farm management. A total of 37 farms were visited in the Mid-South USA and analyzed for the genes 16S rRNA, spaQ (Salmonella spp.), Camp-16S (Campylobacter spp.), tetA, tetB, ermF, ermA, mecA, and intI using quantitative PCR. Additionally, 16S rRNA sequence libraries were created. Overall, it appeared that finisher farms were significantly different from nursery and sow farms in nearly all genes measured and in 16S rRNA clone libraries. Nearly all antibiotic resistance genes were detected in all farms. Interestingly, the mecA resistance gene (e.g. methicillin resistant Staphylococcus aureus) was below detection limits on most farms, and decreased as the pigs aged. Finisher farms generally had fewer antibiotic resistance genes, which corroborated previous phenotypic data; additionally, finisher farms produced a less diverse 16S rRNA sequence library. Comparisons of Camp-16S and spaQ GU (genomic unit) values to previous culture data demonstrated ratios from 10 to 10,000:1 depending on farm type, indicating viable but not cultivatable bacteria were dominant. The current study indicated that swine farm management schemes positively and negatively affect microbial and antibiotic resistant populations in CAFO wastewater which has future "downstream" implications from both an environmental and public health perspective.

Temporal flux and spatial dynamics of nutrients, fecal indicators, and zoonotic pathogens in anaerobic swine manure lagoon water.

Confined animal feeding operations (CAFOs) often use anaerobic lagoons for manure treatment. In the USA, swine CAFO lagoon water is used for crop irrigation that is regulated by farm-specific nutrient management plans (NMPs). Implementation of stricter US environmental regulations in 2013 will set soil P limits; impacting land applications of manure and requiring revision of NMPs. Precise knowledge of lagoon water quality is needed for formulating NMPs, for understanding losses of N and C in ammonia and greenhouse gas emissions, and for understanding risks of environmental contamination by fecal bacteria, including zoonotic pathogens. In this study we determined year-round levels of nutrients and bacteria from swine CAFO lagoon water. Statistical analysis of data for pH, electrical conductivity (EC), inorganic and organic C, total N, water-soluble and total minerals (Ca, Cu, Fe, K, Mg, Mn, P, and Zn) and bacteria (Escherichia coli, enterococci, Clostridium perfringens, Campylobacter spp., Listeria spp., Salmonella spp., and staphylococci) showed that all differed significantly by dates of collection. During the irrigation season, levels of total N decreased by half and the N:P ratio changed from 9.7 to 2.8. Some seasonal differences were correlated with temperature. Total N and inorganic C increased below 19 °C, and decreased above 19 °C, consistent with summer increases in ammonia and greenhouse gas emissions. Water-soluble Cu, Fe, and Zn increased with higher summer temperatures while enterococci and zoonotic pathogens (Campylobacter, Listeria, and Salmonella) decreased. Although their populations changed seasonally, the zoonotic pathogens were present year-round. Increasing levels of E. coli were statistically correlated with increasing pH. Differences between depths were also found. Organic C, total nutrients (C, Ca, Cu, Fe, Mg, Mn, N, P, and Zn) and C. perfringens were higher in deeper samples, indicating stratification of these parameters. No statistical interactions were found between collection dates and depths.