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Tillage systems and fertilizer sources impact soil health and ecosystem services. Rainfall simulations can be used to evaluate tillage and fertilizer management effects on ecosystem services associated with runoff amount and quality. Fixed rate (FR; 57 mm h−1 for 60 minutes) and variable rate (VR; based on the pattern of the most frequent spring rain) simulations were conducted on corn (Zea mays L.) plots supplied with conventional fertilizer (CF; ammonium nitrate [NH4NO3]) or poultry litter (PL) and managed with conventional tillage (CT) or no-tillage (NT) on a Cecil soil (Ultisol) near Watkinsville, Georgia. Simulations were conducted in separate years (FR in 2004 and VR in 2005). Tillage and tillage × fertilizer interactions had significant effects on total runoff and sediment loss under FR but not VR. Under FR, 55.8% of the simulated rainfall was partitioned into runoff from CT compared with 9.7% from NT. Under VR, 42% to 60% was partitioned into runoff with no difference among treatments. Sediment loss was 11-fold greater from CT compared with NT under FR (919 g versus 82 g). Under VR, sediment loss varied between 755 g and 2,174 g with no difference among treatments. Tillage and tillage × fertilizer interactions had significant effects on total load for ammonium-nitrogen (NH4-N) and nitrate-nitrogen (NO3-N) in both methods and dissolved reactive and total phosphorus (DRP and TP) under VR. Fertilizer source affected loads of NH4-N under VR and NO3-N and TP under both methods. Our results confirm that 12 years of continuous NT on a Cecil soil, representative of much of the Southern Piedmont, is far superior to CT for reducing runoff and sediment loss. Larger and more intense storms predicted in future climate projections warrant greater adoption of reduced tillage systems to counteract potential runoff and sediment losses from CT systems.

Flue gas desulfurization gypsum (FGDG) from coal-fired power plants is readily available for agricultural use in many US regions. Broiler litter (BL) provides plant available N, P, and K but can be a source of unwanted As, Cu, and Zn. As a source of Ca and S, FGDG can reduce losses of P and other elements in runoff from BL-amended areas. Rainfall simulation plots (2.0 m) were established on a Piedmont Cecil soil growing 'Coastal' bermudagrass ( L.) for hay. Accumulation and transport of As, Cu, Cd, Cr, Hg, Pb, and Zn were evaluated after annual BL applications (13.5 Mg ha) with four FGDG rates (0, 2.2, 4.5, 9.0 Mg ha) and two FGDG treatments (0 and 9 Mg ha) without BL. Runoff As concentrations were sixfold greater with BL than without ( ≤ 0.01) and were similar to BL with FGDG at 2.2, 4.5 or 9.0 Mg ha ( ≤ 0.10). Runoff concentrations of target elements did not increase where FGDG was applied alone. After three annual applications of FGDG and BL, soil concentrations of As, Cr, Pb, Hg, and Cu were well below levels of environmental concern. Our findings indicate that runoff losses of As from BL application are not reduced with FGDG but support other research indicating no identifiable environmental risks from FGDG beneficial use in agricultural systems.

We tested the null hypothesis that nitrification and the distribution of archaeal and bacterial nitrifying communities would not be impacted by long-term applications of the herbicide, glyphosate, to glyphosate-resistant (gR) and non-glyphosate-resistant corn (nongR) under conventional tillage (CT) and reduced tillage (RT) systems. Bulk and rhizosphere soil nitrification poten-tial rates and quantification of amoA genes of archaeal (AOA) and bacterial (AOB) communities were undertaken. In 2013, the nitrification rate of nongR corn with no glyphosate application treatment associated with bulk and rhi-zosphere soils under RT displayed greater (P < 0.05) nitrification rates than gR corn with glyphosate applications. In 2014, the nitrification rate of the rhizosphere soil under RT in gR corn with no glyphosate application treat-ment was greater (P < 0.05) than other rhizosphere soil treatments. Ratios between archaeal and bacterial amoA genes indicated distinct dominance of AOA communities in the rhizosphere soil. The ratios between AOA and AOB amoA genes in bulk soil were more balanced. Regression analyses indicated more significant correlations between nitrification potential determinations and bulk soil nutrients ratios, nutrient acquisition ratios, and microbial com-munities than between analogous rhizosphere soil characteristics. Compared to non-glyphosate treatments, glyphosate applications appeared to disrupt rhizosphere nutrient, exoenzymatic, and microbial relationships. glyphosate applications had an inconsistent inhibitory effect on the nitrification process. In contrast to bulk soil, rhizosphere soil nitrifying communities were most likely in competition with corn roots for ammonia-n, and were thus dominat-ed by more oligotrophic archaeal nitrifiers compared to bacterial nitrifiers.

Agricultural drainage ditches are conduits between production acreage and receiving aquatic systems. Often overlooked for their mitigation capabilities, agricultural drainage ditches provide an important role for nutrient transformation via microbial metabolism. Variations in ecoenzyme activities have been used to elucidate microbial metabolism and resource demand of microbial communities to better understand the relationship between altered nutrient ratios and microbial activity in aquatic ecosystems. Two agricultural drainage ditches, one in the northeast portion of the Arkansas Delta and the other in the lower Mississippi Delta, were monitored for a year. Sediment samples were collected prior to each ditch being dredged (cleaned), and subsequent post-dredging samples occurred as soon as access was available. Seasonal samples were then collected throughout a year to examine effects of dredging on selected nutrient concentrations and ecoenzymatic activity recovery in drainage ditch sediments. Phosphorus concentrations in sediments after dredging decreased 33–66%, depending on ditch and phosphorus extraction methodology. Additionally, ecoenzymatic activity was significantly decreased in most sediment samples after dredging. Fluorescein diacetate hydrolysis activity, an estimate of total microbial activity, decreased 56–67% after dredging in one of the two ditches. Many sample sites also had significant phosphorus and ecoenzymatic activity differences between the post-dredge samples and the year-long follow-up samples. Results indicate microbial metabolism in dredged drainage ditches may take up to a year or more to recover to pre-dredged levels. Likewise, while sediment nutrient concentrations may be decreased through dredging and removal, runoff and erosion events over time tend to quickly replenish nutrient concentrations in replaced sediments. Understanding nutrient dynamics and microbial metabolism within agricultural drainage ditches is a crucial step toward addressing issues of nutrient enrichment in aquatic receiving systems, especially those contributing to the Gulf of Mexico.

Cryptosporidia are opportunistic pathogens, some of which infect humans when ingested in water or food. Filtration through granular media is the primary mechanism to remove them from water....

BACKGROUND We report results of the last two years of a 7-year field experiment designed to test the null hypothesis: applications of glyphosate on glyphosate resistant (GR) and non-resistant (nonGR) corn (Zea mays L.) under conventional tillage and no-till would have no effect on soil exoenzymes and microbial activity. RESULTS Bulk soil (BS) and rhizosphere soil (RS) macronutrient ratios were not affected by either GR or nonGR corn, or glyphosate applications. Differences observed between exoenzyme activities were associated with tillage rather than glyphosate applications. In 2013 nutrient acquisition ratios for bulk and rhizosphere soils indicated P limitations, but sufficient assimilable N. In 2014 P limitations were observed for bulk and rhizosphere soils, in contrast to balanced C and N acquisition ratios in rhizosphere soils. Stoichiometric relationships indicated few differences between glyphosate and non-glyphosate treatments. Negative correlations between C:P and N:P nutrient ratios and nutrient acquisition ratios underscored the inverse relation between soil nutrient status and microbial community exoenzyme activities. CONCLUSIONS Inconsistent relationships between microbial community metabolic activity and exoenzyme activity indicated an ephemeral effect of glyphosate on BS exoenzyme activity. Except for ephemeral effects, glyphosate applications appeared not to affect the function of the BS and RS exoenzymes under conventional tillage or no-till.

The curve number (CN) method for estimating direct runoff from a rainfall event is commonly used in hydrologic investigations, but currently accepted CNs for no-till cropping might need revising. We derived CNs from ranked rainfall-runoff data gathered from 1972 to 2010 from a 2.7 ha, zero-order watershed (P1) near Athens, Georgia. The watershed was initially managed under conventional tillage for 2.5 years, followed by double-cropped continuous no-till rotations. During conventional tillage, on average 19% of the rainfall was partitioned into runoff (median 13%). The mean and median CN was 82 and 85, respectively. Least square fitting of CN versus rainfall produced an asymptotic CN of 81 (R 2 = 0.61). In contrast, the mean and median runoff during no-till was 7% and 0.6% of the rainfall, respectively. The mean CN equaled the median CN of 62. The asymptotic CN was 58 (R 2 = 0.78), compared with 72 from standard tables. When estimated using CN of 72, the mean runoff during no-till was 142% greater than the measured mean runoff. These results support the hypothesis that CNs for no-till cropping systems should be smaller than currently accepted values for comparable watersheds in the region. This has implications for hydrologic modeling for water quantity and quality where the use of the CN method is ubiquitous.

Storm events that increase flow rates can disturb sediments and produce overland runoff in watersheds with animal agriculture, and, thus, can increase surface water concentrations of fecal bacteria and risk to public health. We tested the hypothesis that strategically designed and placed ponds in watersheds with animal agriculture would attenuate downstream fluxes of fecal bacteria. We measured concentrations and fluxes of fecal indicator bacteria (commensal Escherichia coli and fecal enterococci) and manure pathogens (Salmonella and E. coli 0157:H7) in in- and outflows of Bishop Pond in the Southern Piedmont of Georgia during three storm events and in- and outflow concentrations and fluxes of fecal indicator bacteria at Ponds A and C in the Coastal Plain of Georgia during two storm events. Mean concentrations and fluxes of fecal indicator bacteria associated with pond in- and outflow during hydrograph rise, peak, fall, and 5-days after peak flow at Bishop Pond were significantly greater than their mean base flow concentrations and fluxes. In storm flow Bishop Pond significantly reduced the outflow concentrations and fluxes of fecal indicator bacteria compared with corresponding inflow measurements. Unlike fecal indicator bacteria, Bishop Pond appeared not to reduce outflow concentrations and fluxes of Salmonella or E. coli 0157:H7. At Ponds A and C in the Coastal Plain mean in- and outflow concentrations and fluxes of the fecal indicator bacteria associated with the hydrograph rise and peak flows of the storms were not different. Bishop Pond, with a length to width ratio of 3.3, attenuated downstream fluxes of fecal bacteria. In contrast, Ponds A and C were not effective at reducing downstream fluxes of fecal bacteria under storm flow conditions. The ineffectiveness of Ponds A and C may be attributed to their having length to width ratios of 1.2 and 2.5, respectively, both of which are below the minimum for effective pond performance. Our results indicated that in the humid Southeast an appropriately placed and configured pond in watersheds with animal agriculture can reduce storm flow loads of fecal indicator bacteria but not necessarily pathogenic E. coli 0157:H7.

Georgia farmers are increasing preemergence applications of soil residual herbicides to control glyphosate resistant weeds. To improve efficacy these herbicides are often activated by post-application irrigation. Broiler litter is commonly applied to fields before these herbicides. The herbicide wetting-in practice increases surface soil water content and may increase runoff and transport of broiler litter borne fecal bacteria into surface waters during subsequent storm events. Our objective was to determine differences in loads of fecal bacteria, Escherichia coli and Salmonella spp., in runoff from conventional tillage (CT) and no-till (NT) systems after herbicides were watered into an Ultisol fertilized with broiler litter. On replicated 6 m 2 -plots ( n = 3) simulated rainfall was applied for 70 min with composite runoff samples collected every 5 min and analyzed for E. coli and Salmonella spp. Although total runoff volume from the CT plots was significantly greater than from NT plots, no significant differences in total load of E. coli and Salmonella or the percent of total loads of E. coli and Salmonella recovered in runoff between tillage systems were observed. Total percentage of Salmonella recovered in runoff from both tillage systems was, however, four log 10 orders of magnitude greater than the percentage of E. coli that was recovered. Difference in percentage recovered between the fecal indicator bacterium, E. coli , and the pathogen, Salmonella , underscores an apparent difference in hydrologic transport characteristics of these two fecal bacteria and casts doubts on the efficacy of E. coli as an indicator of risk to public health.

Gypsum has been shown to reduce runoff on soils prone to crust formation in the southeastern United States. Increased infiltration from gypsum applications could therefore help reduce runoff P and other nutrient losses from application of broiler litter (BL), a nutrient-rich fertilizer. In rainfall simulation experiments in June 2009 and May 2011, runoff and nutrient (N, P, Ca, Mg) losses in runoff were compared among treatments consisting of 0, 2.2, 4.5, and 9.0 Mg ha⁻¹ flue gas desulfurization gypsum (FGDG) combined with 13.5 Mg ha⁻¹ of BL and two control treatments of (0–0) and (9.0–0.0) Mg ha⁻¹ (FGDG-BL). A randomized block design with three replications was set up on a Cecil (Typic Kanhapludult) soil growing Coastal bermudagrass (Cynodon dactylon L.) for hay near Watkinsville, Georgia. Amendments were applied each April from 2009 to 2011. A reduction in runoff of 30% each year from the (9.0–13.5) compared with the (0–0) treatment was not statistically significant. Gypsum was effective in reducing concentration and load in one of the two study years: P and NH₄–N in 2009 (up to 83%) and NO₃–N in 2011 (up to 73%). A combination of factors related to weather extremes, timing of FGDG and BL applications, and their implication on soil and vegetation responses at different landscape positions might have caused the different responses between years. Additional studies focused on isolating the impacts of such factors would be helpful to ascertain the effectiveness of multiyear applications of FGDG as a best management practice to reduce P and other nutrient losses in soils of the southeastern United States.

Salmonella and Campylobacter prevalence in stream networks of the Satilla River Basin (SRB) were monitored monthly from August 2007 to August 2009 to study relationships between these pathogens and land use, presence of poultry houses and wastewater treatment plant (WWTP) discharge. Salmonella and Campylobacter were detected at all 10 stream sites and the three sites at the sole wastewater treatment plant (WWTP) in the study area. In all, 43% (129/299) and 62% (96/156) of samples were positive for Salmonella and Campylobacter , respectively, with detection frequency increasing in downstream sites with more poultry production and influence of WWTP discharge. Both Salmonella and Campylobacter detection frequencies were positively associated with the number of poultry houses in the subwatersheds, but agricultural land use as a proportion of the watershed was not a significant predictor of either pathogen. Fecal indicator bacterial levels were assessed and evaluated for their ability to predict the presence of pathogens. Of those examined, enterococci was most predictive; of the 129 samples positive for Salmonella , 88% (113/129) were detected when enterococci were above EPA single sample threshold (61 CFU 100 ml -1 ); and of the 96 samples positive for Campylobacter , 90% (86/96) were detected when enterococci levels exceeded this level. Comparatively, Escherichia coli concentrations were above EPA single sample thresholds in 38% (49/129) of the positive Salmonella samples. Detection of the pathogens throughout the watershed indicated that there was potential for waterborne transmission especially in downstream areas that were more likely to have recreational users.

We report results of the last two years of a 7 year field experiment designed to test the null hypothesis: applications of glyphosate on glyphosate-resistant (GR) and non-resistant (non-GR) corn (Zea mays L.) under conventional tillage and no-till would have no effect on soil exoenzymes and microbial activity.Bulk soil (BS) and rhizosphere soil (RS) macronutrient ratios were not affected by either GR or non-GR corn, or glyphosate applications. Differences observed between exoenzyme activities were associated with tillage rather than glyphosate applications. In 2013, nutrient acquisition ratios for bulk and rhizosphere soils indicated P limitations, but sufficient assimilable N. In 2014, P limitations were observed for bulk and rhizosphere soils, in contrast to balanced C and N acquisition ratios in rhizosphere soils. Stoichiometric relationships indicated few differences between glyphosate and non-glyphosate treatments. Negative correlations between C:P and N:P nutrient ratios and nutrient acquisition ratios underscored the inverse relation between soil nutrient status and microbial community exoenzyme activities.Inconsistent relationships between microbial community metabolic activity and exoenzyme activity indicated an ephemeral effect of glyphosate on BS exoenzyme activity. Except for ephemeral effects, glyphosate applications appeared not to affect the function of the BS and RS exoenzymes under conventional tillage or no-till. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

In the Southeastern Coastal Plain (USA) farmers with access to irrigation commonly water in herbicides following their preemergence application to improve efficacy of the herbicide. This practice increases near surface soil water content and may promote increased runoff during subsequent storm events. The objective of our study was to determine differences in P and N runoff from conventional tillage (CT) and no tillage (NT) systems when herbicides were watered into loamy sand Ultisols fertilized with broiler litter. Simulated rain with variable intensity was applied for 70 min with runoff samples collected every 5 min and analyzed for total P and N, PO 4 3− , NH 4 + and NO 3 − . Results indicated the no-tillage treatment lost significantly more PO 4 3− and NH 4 + than the conventional tillage treatment. However, total P and N and NO 3 − runoff losses were significantly less in the no-tillage treatment. The proportion of total P and N leached through NT residues was 5.6% and 22% of the P and N applied as broiler litter, and the proportion of total P and N in runoff was 0.8% and 1.2% of the P and N applied in broiler litter. Overall 3.4 times more N and 2.7 more P were lost in runoff from the CT treatments than for the NT treatments. Reduced tillage was found to reduce N and P losses when combined with watering-in of surface applied broiler litter.

Animal agriculture in watersheds produces manure bacteria that may contaminate surface waters and put public health at risk. We measured fecal indicator bacteria (commensal Escherichia coli and fecal enterococci) and manure pathogens (Salmonella and E. coli 0157:H7), and physical-chemical parameters in pond inflow, within pond, pond outflow, and pond sediments in three ponds in agricultural watersheds. Bishop Pond with perennial inflow and outflow is located in the Piedmont, and Ponds A and C with ephemeral inflow and outflow in the Coastal Plain of Georgia. Bromide and chloride tracer experiments at Bishop Pond reflected a residence time much greater than that estimated by two models, and indicated that complete mixing within Bishop Pond was never obtained. The long residence time meant that fecal bacteria were exposed to solar UV-radiation and microbial predation. At Bishop Pond outflow concentrations of fecal indicator bacteria were significantly less than inflow concentrations; such was not observed at Ponds A and C. Both Salmonella and E. coli 0157:H7 were measured when concomitant concentrations of commensal E. coli were below the criterion for surface water impairment indicating problems with the effectiveness of indicator organisms. Bishop Pond improved down stream water quality; whereas, Ponds A and C with ephemeral inflow and outflow and possibly greater nutrient concentrations within the two ponds appeared to be less effective in improving down stream water quality.

Poultry (Gallus gallus domesticus L.) litter (PL) is a readily available nutrient source for crop production in the Southeast USA. Long-term PL application may alter availability of N and the effect may be dependent on tillage practice. Tillage [no till (NT) vs. conventional (CT)] and N source (PL vs. commercial fertilizer CF) effects on N availability and plant uptake were evaluated in years 9, 10, and 11 of a long-term cropping systems study at the United States Department of Agriculture, Agricultural Research Service, J. Phil Campbell Sr. Natural Resource Conservation Center, Watkinsville, GA, USA. Mineral N in the top 10 cm, measured in situ, varied each year and was influenced by time, tillage, and N source. In 2003 (year 9), soil mineral N content was greater in CT–CF (100 kg ha−1) than in NT–PL (95 kg ha−1) but in 2004 (year 10) and 2005 (year 11) it was lower in CT–CF (93 and 60 kg ha−1) compared to NT–PL (140 and 71 kg ha−1). Nitrogen mineralization rates were generally greater for PL than for CF treatments with the difference being almost 1 kg ha−1 day−1 in 2003. Mineralization rates were greater for NT–PL compared to CT–CF in 2004 and 2005. Across the three growing seasons, corn (Zea mays L.) aboveground biomass was consistently greater in the NT–PL treatment than in the NT–CF and CT–CF treatments. Correlation between aboveground biomass and N mineralization was greater for PL than for CF (0.75 vs. 0.48). Patterns of N mineralization and total soil mineral N indicated that the distribution of N through the growing season more closely matched corn N demand in PL treatments. Results indicate that improved N availability through the growing season, by combining NT and PL, can result in more profitable corn production in the southeast.

In situ and in vitro experiments were performed to assess the effects of solar radiation and predation by indigenous microflora on the relative die-off rates of a toxigenic strain of Escherichia coli 0157:H7, commensal E. coli, and fecal enterococci in surface waters from ponds in agricultural watersheds. The objective of these experiments was to discern a mechanism of persistence of E. coli 0157:H7 in surface waters compared to fecal indicator bacteria. Results of these experiments indicated that E. coli and fecal enterococci were affected by both insolation and apparent predation; whereas E. coli 0157:H7 appeared to be resistant to both of these environmental stressors. The number of days to reach 99% die-off (T99-values) for E. coli 0157:H7 was significantly greater than that for the indicator bacteria. The capacity to prolong die-off may be connected to the apparent persistence of E. coli 0157:H7 in surface waters.

Submersible pressure transducers integrated with data loggers are commonly used to measure stage (water level) and estimate flow rate from established stage-discharge relationships; however, proper data quality assurance is important to reduce uncertainty in resulting estimates. For 2 yr, USDA-ARS researchers in Watkinsville, Georgia, continuously monitored flow rate from a spring using a flume fitted with a pressure transducer and data logger and performed periodic manual flow rate checks. Initially, during a 162-day period in 2003 with 72 comparisons, the automated system underestimated the mean flow rate by approximately 17% due to presumed drift and lack of frequent calibration. After calibrating the transducer to adjust for drift with the data logger program, 66 comparisons during a 349-day period in 2004 showed that the difference in mean flow rate between the manual and automated estimates was < 5%. During 235 days late in the monitoring period, the automated system overestimated the mean flow rate by approximately 29% because of another apparent drift. In a similar set-up at a pond outflow, a drift of 32 mm in transducer output caused underestimation of pond outflow by 27% during a 30-day period. Users should not assume that pressure transducer-based automated water level measurement systems continuously record correct data. Routine verification of transducer output is crucial to avoid inaccurate (and possibly useless) stage and flow rate estimates.

Conservation tillage and judicious use of animal manures as fertilizers can make significant contributions for sustainable food production in the twenty-first century. Identifying and understanding the many interactions occurring within agricultural systems is fundamental for accomplishing this feat. This paper synthesizes 14 years of research results from a study that began in the early 1990s in which researchers from USDA-ARS and the University of Georgia investigated cropping system influences on nutrient management under natural rainfall. Increases in C and N with no-till resulted in improved soil structure that increased infiltration rate and soil water availability. Biological activity as indicated by earthworms was greater with no-till and poultry litter (PL). In all but the very driest year, yields of cotton and corn increased on average 10–27% with no-till and 32–42% with combination of no-till and PL. On the other hand soil nutrient accumulation, particularly P and Zn from long-term use of poultry litter in corn production, reached excessive levels and could present environmental risks. Drainage increased in no-till raising the risk of leaching of nutrients into the soil profile. However, runoff decreased in no-till and the presence of a rye cover crop during the winter reduced the leaching losses of N compared to no cover crop. During cotton production under relative drought, no-till and poultry litter led to somewhat elevated dissolved phosphorus concentration in runoff, and fluometuron was detected in runoff and drainage while pendimethalin was not. Fecal indicator bacteria (Escherichia coli and fecal enterococci), and the hormones estradiol and testosterone were observed in drainage and runoff but concentrations were similar across all treatments. By conducting the study for an extended period under natural environmental conditions, we were able to highlight real risks and potentials of the contrasting cropping systems. While 6 out of 14 years of relative drought might have limited the water quality response of treatments, such droughts are common features of the weather pattern in the region. Even then, use of no-till as the predominant tillage system was supported by improved yields. Fertilizer management, especially crop N need-based use of PL, requires closer monitoring to insure that production advantages of no-till and poultry litter are not offset by concerns with environmental risks. Long-term research requires sustained resource inputs to answer critical questions of environmental risk and emerging unknown issues.

Poultry litter (PL) can supply N, P, K, and other plant nutrients; however, excessive application may cause environmental problems, depending on management and crop nutrient demand. Changes in soil test (ST) nutrient content in a Cecil soil (a fine, kaolinitic, thermic Typic Kanhapludult) during a 10-yr period of PL use was evaluated at the USDA-ARS J. Phil Campbell, Sr., Natural Resource Conservation Center, Watkinsville, GA. During the cotton (Gossypium hirsutum L.) cropping phase (1995-2000), 4.4 Mg PL ha -1 yr -1 resulted in small changes in ST nutrient content in the surface 15 cm. Differences were observed between tillage treatments, with less accumulation of Ca, Mg, and Mn and greater accumulation of Zn for no-till (NT) than conventional tillage (CT). During the corn (Zea mays L.) cropping phase (2001-2005), average annual PL inputs (11.2 Mg ha -1 ) increased P and Zn contents, with changes being similar for CT and NT. After 10 yr, ST nutrient contents in the surface 15 cm reflected 25, 4, 45, 26, 17, and 97% of the input from PL for P K, Ca, Mg, Mn, and Zn, respectively. Changes in soil profile nutrient content (to a depth of 60 cm) from 1997 to 2005 were predominantly at 0 to 15 cm, where P and Zn increased >200%. Accumulation of Ca, K, P, and Zn at lower depths was also observed. Strategies for increasing nutrient removal following repeated long-term application of PL should be considered to avoid excessive levels of nutrients.

Soil microbial communities under three agricultural management systems (conventionally tilled cropland, hayed pasture, and grazed pasture) and two fertilizer systems (inorganic fertilizer and poultry litter) were compared to that of a w150-y-old forest near Watkinsville, Georgia. Both 16S rRNA gene clone libraries and phospholipid fatty acid (PLFA) analyses indicated that the structure and composition of bacterial communities in the forest soil were significantly different than in the agricultural soils. Within the agricultural soils, the effect of fertilizer amendment on bacterial communities was more dramatic than either land use or season. Fertilizer amendment altered the abundance of more bacterial groups throughout the agricultural soils. In addition, the changes in the composition of bacterial groups were more pronounced in cropland than in pastures. There was much less seasonal variation between the soil libraries. Community-level differences were associated with differences in soil pH, mineralizable carbon and nitrogen, and extractable nutrients. Bacterial community diversity exhibited a complex relationship with the land use intensity in these agro-ecosystems. The pastures had the highest bacterial diversity and could be characterized as having an intermediate degree of intervention compared to low intervention in forest and high intervention in cropland. Changes in bacterial diversity could be attributed to the abundance of a few operational taxonomic units (OTUs). The microdiversity of abundant OTUs in both forest and cropland was consistent with an increase in abundance of many phenotypically similar species rather than a single species for each OTU. Soil microbial communities were significantly altered by long-term agricultural management systems, especially fertilizer amendment, and these results provide a basis for promoting conservation agricultural systems.

Corn (Zea mays L.) producers in the southeastern United States must overcome soil and water limitations to take advantage of the expanding corn market. In this 2001 to 2005 study on a Cecil sandy loam (fine, kaolinitic, thermic Typic Kanhapludult) near Watkinsville, GA, we compared dry land corn biomass and yield under conventional tillage (CT) vs. no-tillage (NT) with ammonium nitrate or sulfate (based on availability) as conventional fertilizer (CF) vs. poultry litter (PL). In a randomized complete block split plot design with three replications, main plots were under tillage and subplots under fertilizer treatments. The cover crop was rye (Secale cereale L.). Over 5 yr, NT and PL increased grain yield by 11 and 18%, respectively, compared with CT and CF. Combined, NT and PL increased grain yield by 31% compared with conventionally tilled and fertilized corn. Similarly, soil water was 18% greater in NT than CT in the 0- to 10-cm depth. In 2 yr of measurements, dry matter of stalks and leaves and leaf area index under PL were an average of 39 and 22% greater, respectively, than under CF during reproduction. Values were 21 and 6% greater, respectively, under NT than CT but during tasseling. Analysis of 70 yr of daily rainfall records showed that supplemental irrigation is needed to meet optimal water requirement. Our results indicate that corn growers can use rainfall more efficiently, reduce yield losses to drought, and expect increased corn yields with a combination of no-tillage management and long-term use of poultry litter.

Aims: To better understand and manage the fate and transport of Salmonella in agricultural watersheds, we developed a culture-based, five tube‐four dilution most probable number (MPN) method for enumerating dilute densities of Salmonella in environmental waters. Methods and Results: The MPN method was a combination of a filtration technique for large sample volumes of environmental water, standard selective media for Salmonella and a TaqMan confirmation step. This method has determined the density of Salmonella in 20-l samples of pond inflow and outflow streams as low as 0AE1 MPN l )1 and a low 95% confidence level 0AE015 MPN l )1 . Salmonella densities ranged from not detectable to 0AE55 MPN l )1 for pond inflow samples and from not detectable to 3AE4 MPN l )1 for pond outflow samples. Salmonella densities of pond inflow samples were associated with densities of Escherichia coli and faecal enterococci that indicated stream contamination with faeces and with nondetectable pond outflow densities of the faecal indicator bacteria. The MPN methodology was extended to flux determinations by integrating with volumetric measurements of pond inflow (mean flux of 2AE 5ls )1 ) and outflow (mean flux of 5AE 6ls )1 ). Fluxes of Salmonella ranged from 100 to greater than 10 4 MPN h )1 . Conclusions: This is a culture-based method that can detect small numbers of Salmonella in environmental waters of watersheds containing animal husbandry and wildlife. Significance and Impact of the Study: Applying this method to environmental waters will improve our understanding of the transport and fate of Salmonella in agricultural watersheds, and can be the basis of valuable collections of environmental Salmonella.

Flue gas desulfurization (FGD) gypsum is a byproduct of coal-fired power plants. Its application to agricultural fields may increase water infiltration, reduce soil erosion, and decrease nutrient losses from applications of animal manures. It may also reduce fecal bacterial contamination of surface waters. We tested the hypothesis that FGD gypsum applications would decrease the load of and the fecal indicator bacterium from poultry litter applications. Two rainfall simulation experiments were undertaken: one in spring 2009 and one in spring 2011. Six treatments consisted of four rates of FGD gypsum (0, 2.2, 4.5, and 9.0 Mg ha) with poultry litter (13.5 Mg ha and two controls) in a randomized, complete-block design with three replications. Each replicate 4- × 6-m plot contained a single 1- × 2-m subplot that was delineated by metal plates and a flume that captured total overland flow or runoff. Rainfall was applied at ∼64 mm h. Volume of overland runoff was measured and subsampled for analysis every 10 min for 1 h. Flow-weighted concentrations, total loads, and soil concentrations of were determined. was not detected in runoff. No significant differences between treatments were observed for the 2009 rainfall simulation. However, after 3 yr of FGD gypsum applications, the highest rate of FGD gypsum resulted in decreased flow-weighted concentrations and total loads of . Flue gas desulfurization gypsum applications may be a management practice that reduces microbial contamination of surface waters from manure applied to agricultural fields in the southeastern United States.

In contrast to endophyte-free (E−) tall fescue, endophyte-infected (E+) tall fescue pastures appear to enhance soil carbon sequestration. A hypothetical mechanism that may account for the enhanced carbon sequestration is that the E+ tall fescue affects the soil microbial community or components of it that are involved in organic carbon turnover. A 60-week mesocosm study with a factorial arrangement of soil type, loamy sand (LS) and clay loam (CL), and E+ and E− tall fescue was conducted to determine if the soil microbial communities were affected by the presence of the endophyte. Bulk and rhizosphere soil samples were fixed in paraformaldehyde, and prepared for total direct microbial counts, and with a combination of one of a domain or subdivision fluorescent oligonucleotide probe for enumerating metabolically active Eubacteria, bacterial subdivisions, and Archaea. E+ tall fescue suppressed the archaeal and high G+C gram-positive bacterial communities of the bulk CL, the delta-proteobacterial community in the rhizosphere CL, and the Planctomycetes community of the rhizosphere LS. In the long-term, suppression of these microbial communities may be a factor in enhanced soil carbon sequestration associated with E+ tall fescue.

Land-applied domestic animal wastes contain appreciable amounts of 17beta-estradiol (henceforth, estradiol) and testosterone. These sex hormones may be transported through soil to groundwater and streams, where they may adversely affect the environment. Previous column transport studies with these hormones used repacked soil and did not consider preferential flow. We, therefore, determined the sorption and transport characteristics of estradiol and testosterone in undisturbed soil columns (15-cm i.d. by 32-cm height). In the sorption experiment, isotherms for estradiol and testosterone were nonlinear with Freundlich exponents (n) less than one. Sorption of both hormones decreased with soil depth, and estradiol sorbed more strongly than testosterone. Average estradiol Freundlich sorption coefficients (K(f)) values were 36.9 microg(1 - n) mL(n) g(-1) for the 0- to 10-cm soil depth and 25.7 microg(1 - n) mL(n) g(-1) for the 20- to 30-cm soil depth. Average testosterone K(f) values were 26.7 microg(1 - n) mL(n) g(-1) for the 0- to 10-cm soil depth and 14.0 microg(1 - n) mL(n) g(-1) for the 20- to 30-cm soil depth. In the transport experiment, 27% of the estradiol and 42% of the testosterone leached through the soil columns. Approximately 50% of the remaining soil-bound hormones were sorbed in the top 10 cm of soil. In almost all instances, breakthrough concentrations of estradiol, testosterone, and a chloride tracer peaked simultaneously. Simultaneous breakthrough and HYDRUS-1D transport parameters indicated both chemical and physical nonequilibrium processes affected hormone transport. This suggests hormones placed on soil surfaces may contaminate groundwater under conditions of preferential flow.

The application of poultry litter to agricultural fields can provide plant nutrients for crops and forage production, but fecal bacteria and the sex hormones estradiol and testosterone are components of litter that can be detrimental to the environment. Our objective was to determine if applications of poultry litter to small watersheds would contribute to the load of fecal bacteria and sex hormones to soil and runoff. We, therefore, investigated the fate and transport of fecal bacteria, estradiol and testosterone from surface applied poultry litter to four small watersheds. Poultry litter was applied to meet the nitrogen requirements of pearl millet (Pennisetum glaucum L.) in 2000 and grain sorghum [Sorgham bicolor (L.) Moench] in 2001. Neither Salmonella nor Campylobacter were detected in the litter but the fecal indicator bacteria were. The average load of total coliforms,Escherichia coli, and fecal enterococci applied with the litter was 12.2, 11.9, and 12.7 log10 cells ha(−1), respectively. The average load of estradiol and testosterone was 3.1 and 0.09 mg ha(-1), respectively.Runoff events first occurred seven months after the first litter application in 2000, and three weeks after the second application in 2001.Only for the 25 July 2001 runoff event three weeks after the second litter application, were the concentrations of total coliforms, E. coli,and fecal enterococci in runoff greater than background concentrations which were on average 5.2, 1.1, and 2.9 log10 MPN 100 ml(−1),respectively [corrected]. Average background levels of total coliforms, fecal enterococci,and E. coli in surface soil were 8.2, 7.9, and 3.5 log (10) cells kg(−1) soil. At the rate of litter application the concentrations of estradiol and testosterone in the litter did not appear to impact the background levels in the soil and runoff. Because concentrations of sex hormones in litter from other broiler operations are known to be greater than in the litter we applied, further study on the connection between concentrations of sex hormones in poultry litter and operational practices is recommended.

The effects of liquid CO2 injection on the viability of Cryptosporidium parvum oocysts were evaluated. A laboratory study was designed to test the effects of saturated CO2, freeze–thaw cycles and different freezing protocols on C. parvum oocysts in aquifer material. Oocysts were exposed to a saturated solution of CO2 at room temperature for 1-, 4-, 8-, and 12-h intervals and their viability was compared with controls. One- and three-cycle freeze–thaw experiments on oocyst survival were conducted. Inactivation of oocysts was assessed for: (1) rapid freezing and rapid thawing and (2) gradual freezing and rapid thawing. Exposure to 1 atm of CO2 in water at room temperature had a negligible effect on oocyst viability. Average oocyst viability after the one- and three-cycle freeze–thaw experiments was 24.7 and 2.7%, repsectively. The average oocyst viability associated with the rapid freeze–thaw and gradual freeze–thaw experiments was 11.3 and 26.2%, respectively. Freezing associated with injection of liquid C...

Rhizobial symbionts were isolated from the surface (0–0.5 M) and phreatic (3.9–5.0 M) root environments of a mature mesquite woodland in the Sonoran Desert of Southern California, and from variable depths (0–12 m) of non-phreatic mesquite ecosystems in the Chihuahuan Desert of New Mexico. They were tested for their ability to tolerate high salinity, and respire NO 3 − as mechanisms of free-living survival. Sixteen of 25 isolates were grown in yeast-extract mannitol (YEM) broth at NaCl concentrations of 2 (basal concentration), 100, 300, 500 and 600 mM, and their specific growth rates, cell dry weight and lag times were determined. Twenty of the 25 isolates were also grown in YEM broth under anaerobic conditions with or without 10 mM KNO 3 . Three categories of NaCl salinity responses were observed: (1) eight isolates showed decreased specific growth rates at NaCl concentrations of 100, 300 and 500 mM, but they nevertheless remained viable at 500 mM NaCl concentration; (2) the specific growth rate of six isolates increased significantly at 100 and 300 mM NaCl; and (3) specific growth rates of two isolates were significantly greater than the base-rate at all concentrations of NaCl. Five of 11 of the Bradyrhizobium isolates tested respired NO 3 − , but showed no growth. Seven Rhizobium isolates, three from the deep (3.9–5 m) phreatic rhizobial community, and four from the surface community denitrified NO 3 − but only the isolates from the phreatic community displayed anaerobic growth. Long-term interactions between rhizobial and bradyrhizobial communities and the surface and phreatic root environments of the mature Sonoran Desert mesquite woodland appear to have selected for strains of NO 3 − respiring rhizobia, general salt tolerance of both rhizobial and bradyrhizobial symbionts, and strains of weak facultative halophilic bradyrhizobia. These survival characteristics of mesquite rhizobia may be important regarding mesquite's establishment and long-term productivity in marginal desert soils, and may provide novel types of rhizobia for food crops growing in harsh environments.

Escherichia coli is a ubiquitous component of the intestinal microflora of warm-blooded animals, and is an indicator of fecal contamination of surface waters. Ribotype profiling of E. coli is one of several genotypic methods that has been developed to determine the host origin of fecal bacteria. Like most genotypic methods of source tracking, ribotyping requires a host origin database to identify environmental isolates. To determine the extent of temporal variability of ribotypes and its effect on a host origin database, E. coli isolates were obtained from fecal samples of two herds of Black Angus steers at a long-term experimental site at four sampling times from October 1999 to July 2000. Fecal samples were taken from six randomly chosen steers at each time. At a similarity index of 90% as calculated by unweighted pair-group method using arithmetic averages (UPGMA), 240 ribotypes were identified from 451 E. coli isolates. Only 20 ribotypes (8.3%), comprising 33% of the total isolates, were shared among sampling times and were considered resident ribotypes. Two of the twenty resident ribotypes appeared at three sampling times, and the remaining eighteen appeared at two. The majority of the ribotypes, therefore, were transient and unique to each sampling time and steer. Both the apparent turnover of E. coli ribotypes and a clonal diversity index of 0.97 (indicative of extensive ribotype variability) suggest the necessity of ribotyping a large number E. coli isolates per host to establish a host origin database that is independent of temporal variability, or complete enough to be effective.

The interaction between soil types, temperature, and soil water potential may have differential effects on the survival of Cryptosporidium parvum oocysts in the terrestrial environment. We examined the effects of three soil types (a silty clay loam, silt loam, and loamy sand), three temperatures (4, 20, and 30 °C), and three soil water potentials (−0.033, −0.5 and −1.5 MPa) on the inactivation kinetics of oocysts. Sentinel chambers were filled with air-dried and sieved soil, brought to the appropriate soil water potential, and inoculated with 2×106 freshly purified oocysts. The inoculated chambers were buried in the same bulk soil at the appropriate water potentials and incubated at one of the three temperatures. Triplicate chambers were removed from the bulk soil on days 0, 22, 43, 84 and 156. Sentinel oocysts were extracted, and assayed for potential infectivity by the dye permeability method. Oocysts suspended in sterile distilled water and incubated with the sentinel chambers were used as controls for the effect of temperature. The soil water potentials investigated did not affect oocyst inactivation at any temperature or with any of the three soil types. Rates of oocyst inactivation increased significantly between 4 and 20 °C, but not between 20 and 30 °C with the exception of oocysts incubated in the silty clay loam. Oocyst survival appeared to be significantly greater in the silt loam soil than in the two other soil types when incubated at 20 °C; and at 30 °C oocyst survival was significantly less in the silt clay loam than in the other two soil types. Rates of sentinel oocyst inactivation at all three soil water potentials were significantly lower than the control oocysts in water at the three test temperatures. Thus oocyst survival in soil was not affected by the water potentials between −0.033 and −1.5 MPa; it was affected by soil texture; but temperature appeared to be the factor most affecting oocyst survival. In the critical ambient range of temperature in temperate climates oocysts may survive for months in agricultural soil, and pose a threat to surface waters.

The effects of freeze-thaw events on the inactivation of Cryptosporidium parvum oocysts in soil were examined. Oocysts were inoculated into distilled water in microcentrifuge tubes or into chambers containing soil the water content of which was maintained at 3%, 43%, or 78% of the container capacity. The chambers and tubes were then embedded in 3 soil samples from different aspects of a hillside landscape (Experiments 1 and 2) and in 3 distinct soil types (Experiment 3) and frozen at -10 C. Containers were thawed every 3 days for a period of 24 hr in 1-9 freeze-thaw cycles over 27 days (Experiments 1 and 2) and 2-5 freeze-thaw cycles over 15 days (Experiment 3). Oocyst viability was measured using the fluorescent dyes 4'6-diaminidino-2-phenylindole and propidium iodide. Inactivation rates were greater in soils than in water and greater in dry soil than in moist and wet soils. Soil type showed no effect on inactivation. Oocysts subjected to freeze-thaw cycles had inactivation rates not significantly different from those of oocysts subjected to -10 C under static conditions. The results indicated that 99% of oocysts exposed to soils that are frozen at -10 C will become inactivated within 50 days whether or not freeze-thaw cycles occur.

Calves make up about 16% of the current bovine population in the United States and can excrete high levels of human pathogens in their feces. We describe the density and distribution of genetic markers from 9 PCR- and real-time quantitative PCR-based assays, including CF128, CF193, CowM2, CowM3, GenBac3, Entero1, EC23S857, CampF2, and ttr-6, commonly used to help assess ambient surface water quality. Each assay was tested against a collection of 381 individual bovine fecal samples representing 31 mother and calf pairings collected over a 10-month time period from time of birth through weaning. Genetic markers reported to be associated with ruminant and/or bovine fecal pollution were virtually undetected in calves for up to 115 days from birth, suggesting that physiological changes in calf ruminant function impact host-associated genetic marker shedding. In addition, general fecal indicator markers for Bacteroidales , Escherichia coli , and Enterococcus spp. exhibited three separate trends across time, indicating that these bacteria respond differently to age-related physiological and dietary changes during calf development. The results of this study suggest that currently available PCR-based water quality indicator technologies can under- or overestimate fecal pollution originating from calves and identify a need for novel calf-associated source identification methods.

Cryptosporidium parvum oocysts were examined to ascertain excystation requirements and the effects of gamma irradiation. Oocysts and excysted sporozoites were examined for dye permeability and infectivity. Maximum excystation occurred when oocysts were pretreated with acid and incubated with bile salts, and potassium or sodium bicarbonate. Pretreatment with Hanks' balanced salt solution or NaCl lowered excystation; however, this effect was overcome with acid. Sodium ions were replaceable with potassium ions, and sodium bicarbonate was replaceable with sodium phosphate. Oocysts that received 200 krad irradiation excysted at the same rates as nonirradiated oocysts (95%), the excystation rates were lowered (50%) by 2,000 krad, and no excystation was observed by 5,000 krad. No differences were observed between the propidium iodide (PI) permeability of untreated oocysts and oocysts treated with 200 krad, while 92% of oocysts were PI positive after 2,000 krad. Most of the sporozoites exposed to 2,000 krad were not viable as indicated by the dye permeability assay. The oocysts irradiated with 200 and 2,000 krad infected cells, but no replication was observed. The results suggest that gamma-irradiated oocysts may still be capable of excystation and apparent infection; however, because the sporozoites could not reproduce they must not have been viable. The identification of the mechanisms that trigger and carry out the process of excystation is critical for understanding how sporozoites are freed from oocysts prior to initiating the pene- tration of the hosts' intestinal cells. In vitro excystation is a parameter that has been used to measure the viability of spo- rozoites within oocysts (Campbell et al., 1992; Jenkins et al., 1997). Jenkins et al. (1997) reported that the in vitro excystation assay for potential infectivity of oocysts correlated with the in vivo mouse infectivity test and the in vitro dye permeability assay. Several studies have examined the requirements for Crypto- sporidium parvum oocyst excystation (Fayer and Leek, 1984

Amendments to the Safe Drinking Water Act (PL-930123) in 1996 required that public water supply managers identify potential sources of contamination within contributing areas. Nonpoint sources of acutely toxic microbial contaminants, such as Cryptosporidium parvum, challenge current approaches to source identification and management as a first step toward developing management plans for public water supply protection. Little may be known about survival and transport in the field environment, prescribed practices may not be designed to manage such substances, and infective stages may be present in vast numbers and may resist water treatment and disinfection processes. This review summarizes research related to survival and transport of C. parvum oocysts, as an example of an acutely toxic contaminant with nonpoint sources in animal agriculture. It discusses ∥1) significance of infected domesticated animals as potential sources of C. parvum, (2) laboratory and field studies of survival and transport, and (3) approaches to source control in the context of public health protection.

The ability to determine inactivation rates of Cryptosporidium parvum oocysts in environmental samples is critical for assessing the public health hazard of this gastrointestinal parasite in watersheds. We compared a dye permeability assay, which tests the differential uptake of the fluorochromes 4'-6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) by the oocysts (A. T. Campbell, L. J. Robertson, and H. V. Smith, Appl. Environ. Microbiol. 58:3488-3493, 1992), with an in vitro excystation assay, which tests their ability to excyst and, thus, their metabolic potential and potential for infectivity (J.B. Rose, H. Darbin, and C.P. Gerba, Water Sci. Technol. 20:271-276, 1988). Formaldehyde-fixed (killed) oocysts and untreated oocysts were permeabilized with sodium hypochlorite and subjected to both assays. The results of the dye permeability assays were the same, while the excystation assay showed that no excystation occurred in formaldehyde-fixed oocysts. This confirmed that oocyst wall permeability, rather than metabolic activity potential, was the basis of the dye permeability viability assessment. A previously developed protocol (L. J. Anguish and W. C. Ghiorse, Appl. Environ. Microbiol. 63:724-733, 1997) for determining viability of oocysts in soil and sediment was used to examine further the use of oocyst permeability status as an indicator of oocyst viability in fecal material stored at 4 degrees C and in water at various temperatures. Most of the oocysts in fresh calf feces were found to be impermeable to the fluorochromes. They were also capable of excystation, as indicated by the in vitro excystation assay, and were infective, as indicated by a standard mouse infectivity assay. The dye permeability assay further showed that an increase in the intermediate population of oocysts permeable to DAPI but not to PI occurred over time. There was also a steady population of oocysts permeable to both dyes. Further experiments with purified oocysts suspended in distilled water showed that the shift in oocyst populations from impermeable to partially permeable to fully permeable was accelerated at temperatures above 4 degrees C. This sequence of oocyst permeability changes was taken as an indicator of the oocyst inactivation pathway. Using the dye permeability results, inactivation rates of oocysts in two fecal pools stored in the dark at 4 degrees C for 410 and 259 days were estimated to be 0.0040 and 0.0056 oocyst day-1, respectively. The excystation assay gave similar inactivation rates of 0.0046 and 0.0079 oocyst day-1. These results demonstrate the utility of the dye permeability assay as an indicator of potential viability and infectivity of oocysts, especially when combined with improved microscopic methods for detection of oocysts in soil, turbid water, and sediments.

Because of outbreaks of cryptosporidiosis in humans, some Cryptosporidium spp. have become a public health concern. Commercial swine operations can be a source of this protozoan parasite. Although the species distribution of Cryptosporidium is likely dominated by Cryptosporidium suis , a fraction may be comprised of other Cryptosporidium species infectious to humans such as Cryptosporidium parvum . To better understand the survival dynamics of Cryptosporidium spp., oocysts associated with swine operations, 2 experiments were performed to determine die-off rates of C. parvum oocysts in a swine waste lagoon (2009 and 2010) and its spray field (2010 and 2011). Sentinel chambers containing a lagoon effluent suspension of C. parvum oocysts were submerged in the lagoon, and triplicate chambers were removed over time; oocysts were extracted and assayed for viability. For comparative purposes, inactivation rates of Ascaris suum eggs contained in sentinel chambers were also determined. For 2 spray field experiments, air-dried and sieved surface soil was placed in sentinel chambers, hydrated, and inoculated with a lagoon effluent suspension of C. parvum oocysts. Sentinel chambers and control oocysts in PBS contained in microcentrifuge tubes were buried 1.5 cm below the soil surface in 3 blocks. Triplicate chambers and controls were removed over time; oocysts were extracted and assayed for viability. Based on the first order decay equation, days to reach 99% die-off (T(99)) were determined. T(99)-values determined for the 2 lagoon experiments were 13.1 and 20.1 wk, respectively. A T(99)-value for C. parvum in the spray field was significantly longer at 38.0 wk than the control oocysts in PBS at 29.0 wk. The waste lagoon and spray field system of manure management at this large-scale farrowing operation appeared to reduce the load of C. parvum oocysts before they can be hydrologically transported off the operation and reduces their likelihood of contaminating surface waters and threatening public health.

Approximately 11% of the Southern Piedmont (1.8 million ha) is used for pasture and hay production, mostly under low-input management. Few studies have investigated in the region long-term nitrogen and carbon losses in surface runoff, which can be significant. We present 1999 to 2009 hydrologic and water quality data from a rotationally grazed, 7.8-ha, zero-order pasture (W1) near Watkinsville in the Georgia Piedmont. Annual rainfall was 176 to 463 mm below the long-term average (1240 mm) in 7 of the 11 yr. There were 20 runoff events during 86 mo of below-average rainfall (deficit period), compared with 54 events during 46 mo of nondeficit period. Mean event flow-weighted concentration (in mg L) was 0.96 for nitrate-nitrogen (NO-N), 0.97 for ammonium-nitrogen (NH-N), 3.70 for total nitrogen (TN), and 9.12 for total organic carbon (TOC) ( = 43-47; limited due to instrument problem). Nutrient loads (in kg ha per event) averaged 0.04 for NO-N, 0.03 for NH-N, 0.19 for TN, and 0.54 for TOC. Total loads for N and TOC were 6 to 11 times greater from nondeficit than from deficit periods. The observed N concentrations, while well below maximum drinking water standard limits, could pose risk for eutrophication, which can be stimulated at lower concentrations. However, the ability of headwater streams, such as the one downstream of W1, to reduce nutrient concentrations might partially alleviate this concern. The results of this study point to the need to use a long-term dataset that includes measurements made in drought and wet years when evaluating the efficacy of water quality standards.

The fecal microbiome of cattle plays a critical role not only in animal health and productivity but also in food safety, pathogen shedding, and the performance of fecal pollution detection methods. Unfortunately, most published molecular surveys fail to provide adequate detail about variability in the community structures of fecal bacteria within and across cattle populations. Using massively parallel pyrosequencing of a hypervariable region of the rRNA coding region, we profiled the fecal microbial communities of cattle from six different feeding operations where cattle were subjected to consistent management practices for a minimum of 90 days. We obtained a total of 633,877 high-quality sequences from the fecal samples of 30 adult beef cattle (5 individuals per operation). Sequence-based clustering and taxonomic analyses indicate less variability within a population than between populations. Overall, bacterial community composition correlated significantly with fecal starch concentrations, largely reflected in changes in the Bacteroidetes , Proteobacteria , and Firmicutes populations. In addition, network analysis demonstrated that annotated sequences clustered by management practice and fecal starch concentration, suggesting that the structures of bovine fecal bacterial communities can be dramatically different in different animal feeding operations, even at the phylum and family taxonomic levels, and that the feeding operation is a more important determinant of the cattle microbiome than is the geographic location of the feedlot.

In situ and in vitro experiments were performed to assess the effects of solar radiation and predation by indigenous microflora on the relative die-off rates of a toxigenic strain of Escherichia coli O157:H7, commensal E. coli, and fecal enterococci in surface waters from ponds in agricultural watersheds. The objective of these experiments was to discern a mechanism of persistence of E. coli O157:H7 in surface waters compared to fecal indicator bacteria. Results of these experiments indicated that E. coli and fecal enterococci were affected by both insolation and apparent predation; whereas E. coli O157:H7 appeared to be resistant to both of these environmental stressors. The number of days to reach 99% die-off (T(99)-values) for E. coli O157:H7 was significantly greater than that for the indicator bacteria. The capacity to prolong die-off may be connected to the apparent persistence of E. coli O157:H7 in surface waters.

Ascaris suum eggs were inactivated in distilled water and digested sludge by butanoic, pentanoic, and hexanoic acids. The fatty acids (short-chain fatty acids [SCFA]) were effective only when protonated and at sufficient concentrations. The conjugate bases were not effective at the concentrations evaluated. Predictions from an inhibition model (50% inhibitory concentration [IC 50 ]) based on quantitative structure-activity relationships were congruent with inactivation data.

Waste lagoons of swine operations are a source of Cryptosporidium oocysts. Few studies, however, have reported on oocyst concentrations in swine waste lagoons; none have reported on oocyst viability status, nor has there been a systematic assessment of species/genotype distributions across different types of swine facilities. Ten swine waste lagoons associated with farrowing, nursery, finishing, and gestation operations were each sampled once a month for a year. Oocysts were extracted from triplicate 900-ml effluent samples, enumerated by microscopy, and assessed for viability by dye exclusion/vital stain assay. DNA was extracted from processed samples, and 18S ribosomal DNA (rDNA) genes were amplified by PCR and sequenced for species and genotype identification. Oocysts were observed at each sampling time at each lagoon. Annual mean concentrations of total oocysts and viable oocysts ranged between 24 and 51 and between 0.6 and 12 oocysts ml −1 effluent, respectively. The species and genotype distributions were dominated (95 to 100%) by Cryptosporidium suis and Cryptosporidium pig genotype II, the latter of which was found at eight of the lagoons. The lagoon at the gestation facility was dominated by Cryptosporidium muris (90%), and one farrowing facility showed a mix of pig genotypes, Cryptosporidium muris , and various genotypes of C. parvum . The zoonotic C. parvum bovine genotype was observed five times out of 407 18S rDNA sequences analyzed. Our results indicate that pigs can have mixed Cryptosporidium infections, but infection with C. suis is likely to be dominant.

There are numerous PCR-based assays available to characterize bovine fecal pollution in ambient waters. The determination of which approaches are most suitable for field applications can be difficult because each assay targets a different gene, in many cases from different microorganisms, leading to variation in assay performance. We describe a performance evaluation of seven end-point PCR and real-time quantitative PCR (qPCR) assays reported to be associated with either ruminant or bovine feces. Each assay was tested against a reference collection of DNA extracts from 247 individual bovine fecal samples representing 11 different populations and 175 fecal DNA extracts from 24 different animal species. Bovine-associated genetic markers were broadly distributed among individual bovine samples ranging from 39 to 93%. Specificity levels of the assays spanned 47.4% to 100%. End-point PCR sensitivity also varied between assays and among different bovine populations. For qPCR assays, the abundance of each host-associated genetic marker was measured within each bovine population and compared to results of a qPCR assay targeting 16S rRNA gene sequences from Bacteroidales . Experiments indicate large discrepancies in the performance of bovine-associated assays across different bovine populations. Variability in assay performance between host populations suggests that the use of bovine microbial source-tracking applications will require a priori characterization at each watershed of interest.

Submersible pressure transducers integrated with data loggers have become relatively common measuring devices in flow or well water elevation measurements. However, drift, linearity, hysteresis and other problems can lead to erroneous data. Researchers at the USDA-ARS in Watkinsville, GA, carried out over a two-year period continuous monitoring of flow from a spring using a flume fitted with a pressure transducer and data logger. Over the same period, periodic manual checking of flow rate was performed. Initially, over a 162-day period in 2003 with 77 comparisons, the automated system under measured the flow rate by approximately 17% (mean, 0.383 versus 0.459 L s-1; significant based on a Wilcoxon Rank Sum Test). Over a 349-day period in 2004 in 66 days of comparison, flow rate was 0.294 L s-1 for the manual and 0.299 L s-1 for the automated system (a 1.7% difference; non-significant). Over 235 days during the latter part of the monitoring period, the automated system over measured the flow rate by approximately 29% (mean 0.599 versus 0.463 L s-1; significant). In a similar set up at a pond outflow, an offset of 32 mm in flow depth caused a negative outflow error of 27.4% (13,141 m3) over a 30-day period. Flow rate is an important hydrologic variable in water quantity and quality investigations. Proper data quality assurance is needed in flow rate estimation utilizing pressure transducers to avoid the propagation of errors from flow estimation to that of pollutant load. The same principle should apply in other similar sensors.

U.S. Army Iodine Water Purification Tablets were tested to determine their efficacy against Cryptosporidium parvum, a protozoan resistant to chemical disinfection. Purified oocysts in phosphate-buffered water were treated with varying concentrations of iodine or with iodine tablets as per U.S. Army protocol. Neonatal mouse pups were then each inoculated with 10,000 treated oocysts, and 1 week later scored as infected or uninfected. Using this methodology, iodine tablets were found to be inadequate against C. parvum because the Army doctrinal dose of 560 mg min/L, calculated as 16 mg of I2/L and 35 minutes of contact time, showed less than 1 log inactivation. A dose of 29 mg of I2/L at the same contact time was required to achieve a 2 log inactivation.