Source availability and hydrological connectivity determined nitrate-discharge relationships during rainfall events in karst catchment as revealed by high-frequency nitrate sensing.

Karst terrain seasonal monsoonal rainfall is often associated with high concentrations of nitrate-N in streams draining agricultural land. Such high concentrations can pose problems for environmental and human health. However, the relationship between rainfall events that mobilize nitrate and resulting nitrate export remains poorly understood in karst terrain. To better understand the processes that drive nitrate dynamics during rainfall events, the characteristics of individual rainfall events were analyzed using sensor technology. Thirty-eight rainfall events were separated from the high-frequency dataset spanning 19 months at a karst spring site. The results revealed that nitrate-discharge (N-Q) hysteresis in 79% of rainfall events showed anticlockwise hysteresis loop patterns, indicating nitrate export from long distances within short event periods. Karstic hydrological connectivity and source availability were considered two major determining factors of N-Q hysteresis. Gradual increase in hydrological connectivity during intensive rainfall period accelerated nitrate transportation by karst aquifer systems. Four principal components (PCs, including antecedent conditions PC1&3 and rainfall characteristics PC2&4 explained 82% of the cumulative variance contribution to the rainfall events. Multiple linear regression of four PCs explained more than 50% of the variation of nitrate loading and amplitude during rainfall events, but poorly described nitrate concentrations and hydro-chemistry parameters, which may be influenced by other factors, e.g., nitrate transformation, fertilization time and water-rock interaction. Although variation of N concentration during event flow is evident, accounting for antecedent conditions and rainfall factors can help to predict rainfall event N loading during rainfall events. Pollution of the karstic catchment occurred by a flush of nitrate input following rainfall events; antecedent and rainfall conditions are therefore important factors to consider for the water quality management. Reducing source availability during the wet season may facilitate to reduction of nitrogen loading in similar karst areas.

Anthropogenic land use and urbanization alter the dynamics and increase the export of dissolved carbon in an urbanized river system.

Greenhouse gas emissions from urban rivers play a crucial role in global carbon (C) cycling, this is tightly linked to dissolved C in rivers but research gaps remain. The effects of urbanization and anthropogenic land-use change on riverine dissolved carbon dynamics were investigated in a temperate river, the River Kelvin in UK. The river was constantly a source of methane (CH4) and carbon dioxide (CO2) to the atmosphere (excess concentration of CH4 ranged from 13 to 4441 nM, and excess concentration of CO2 ranged from 2.6 to 230.6 µM), and dissolved C concentrations show significant spatiotemporal variations (p < 0.05), reflecting a variety of proximal sources and controls. For example, the concentration variation of dissolved CH4 and dissolved CO2 were heavily controlled by the proximity of coal mine infrastructure in the tributary near the river head (~ 2 km) but were more likely controlled by adjacent landfills in the midstream section of the rivers main channel. Concentration and isotopic evidence revealed an important anthropogenic control on the riverine export of CO2 and dissolved organic carbon (DOC). However, dissolved inorganic carbon (DIC) input via groundwater at the catchment scale primarily controlled the dynamics of riverine DIC. Furthermore, the positive relationship between the isotopic composition of DIC and CO2 (r = 0.79, p < 0.01) indicates the DIC pool was at times also significantly influenced by soil respiratory CO2. Both DIC and DOC showed a weak but significant correlation with the proportion of urban/suburban land use, suggesting increased dissolved C export resulting from urbanization. This research elucidates a series of potentially key effects anthropogenic activities and land-use practices can have on riverine C dynamics and highlights the need for future consideration of the direct effects urbanization has on riverine C dynamics.

Rainfall and conduit drainage combine to accelerate nitrate loss from a karst agroecosystem: Insights from stable isotope tracing and high-frequency nitrate sensing.

Understanding where nitrate is mobilized from and under what conditions is required to reduce nitrate loss and protect water quality. Low frequency sampling may inadequately capture hydrological and biogeochemical processes that will influence nitrate behavior. We used high-frequency isotope sampling and in-situ nitrate sensing to explore nitrate export and transformation in a karst critical zone. Nitrate was mobilised during light rainfall, and transferred from soil layers to the karst matrix, where some nitrate was retained and denitrified. Nitrate isotopic composition changed rapidly during the rising limb of events and slowly during the falling limb. The main nitrate source was synthetic fertiliser (up to 80% during event flow), transported by conduit flow following high rainfall events, and this contribution increased significantly as discharge increased. Soil organic nitrogen contribution remained constant indicating at baseflow this is the primary source. Isotope source appointment of nitrate export revealed that synthetic fertilizer accounted for more than half of the total nitrate export, which is double that of the secondary source (soil organic nitrogen), providing valuable information to inform catchment management to reduce nitrate losses and fluvial loading. Careful land management and fertilizer use are necessary to avoid nitrate pollution in the karst agroecosystem, for example by timing fertilizer applications to allow for plant uptake of nitrate before rainfall can flush it from the soils into the karst and ultimately into catchment drainage.

Chronic urban hotspots and agricultural drainage drive microbial pollution of karst water resources in rural developing regions.

Contamination of surface and groundwater systems with human and animal faecal matter leads to exposure of reliant populations to disease causing micro-organisms. This exposure route remains a major cause of infection and mortality in developing countries, particularly rural regions. To meet the UN's sustainable development goal 6: Ensure availability and sustainable management of water and sanitation for all, we need to identify the key controls on faecal contamination across relevant settings. We conducted a high-resolution spatial study of E. coli concentration in catchment drainage waters over 6 months in a mixed land-use catchment in the extensive karst region extending across impoverished southwest China. Using a mixed effects modelling framework, we tested how land-use, karst hydrology, antecedent meteorological conditions, agricultural cycles, hydrochemistry, and position in the catchment system affected E. coli concentrations. Land-use was the best predictor of faecal contamination levels. Sites in urban areas were chronically highly contaminated, but water draining from agricultural land was also consistently contaminated and there was a catchment wide pulse of higher E. coli concentrations, turbidity, and discharge during paddy field drainage. E. coli concentration increased with increasing antecedent rainfall across all land-use types and compartments of the karst hydrological system (underground and surface waters), but decreased with increasing pH. This is interpreted to be a result of processes affecting pH, such as water residence time, rather than the direct effect of pH on E. coli survival. Improved containment and treatment of human waste in areas of higher population density would likely reduce contamination hotspots, and further research is needed to identify the nature and distribution of sources in agricultural land.

Coupled hydrological and biogeochemical modelling of nitrogen transport in the karst critical zone.

Transport of nitrogen (N) in karst areas is more complex than in non-karst areas due to marked heterogeneity of hydrodynamic behaviour in the karst critical zone. Here, we present a novel, distributed, coupled hydrological-biogeochemical model that can simulate water and nitrogen transport in the critical zone of karst catchments. This new model was calibrated using integrated hydrometric, water stable isotope, and nitrogen-N concentration data at the outflow of Houzhai catchment in Guizhou province of Southwest China. Hydrological dynamics appears to control N load from the study catchment. Combining flow discharge and water stable isotopes significantly constrained model parameterisation and mitigate the equifinality effects of parameters on the simulated results. Karst geomorphology and land use have functional effects on spatiotemporal variations of hydrological processes and nitrogen transport. In the study catchment, agricultural fertilizer was the largest input source of N, accounting for 86% of the total. Plant uptake consumed about 45% of inputs, primarily in the low-lying valley bottom areas and the plain covered by relatively thick soils. Thus, a large amount of N released from soil reservoirs to the epikarst (via fractures or sinkholes) is then exported to the underground channel in the limestone area to the south. This N draining into groundwater could lead to extensive, potentially long-term contamination of the karst system. Therefore, improving the efficiency of fertilization and agricultural management in valleys/depressions is an urgent need to reduce N losses and contamination risk.

High-frequency monitoring reveals how hydrochemistry and dissolved carbon respond to rainstorms at a karstic critical zone, Southwestern China.

Hydrochemical behavior and dissolved carbon dynamics are highly-sensitive to hydrological variations in the monsoon-influenced karstic critical zone which has high chemical weathering rates and experiences strong anthropogenic impact. Continuous high-frequency monitoring in the spring outlet of a karstic catchment in Southwestern China revealed that most hydrochemical variables changed distinctively in response to hydrologic variations, influenced by mixing of different sources and miscellaneous biogeochemical processes. Na+, K+ and SO42- varied significantly with hydrology, showing weak chemostatic behavior controlled by dilution. The flushing effect and random behavior of NO3- and Cl- likely reflect agricultural inputs from high throughflow. Soil CO2 in infiltrated water supports carbonate weathering, enabling DIC (dissolved inorganic carbon) and weathering products (e.g., Ca2+ and Mg2+) to maintain chemostatic behavior. Biogenic DIC exhibited a stronger chemostatic response than carbonate sources and was the foremost control in DIC behavior. Carbon exchange between DIC and DOC (dissolved organic carbon) did not significantly influence DIC concentration and δ13C due to very low DOC concentration. More DOC was exported by flushing from increasing discharge. Hysteretic analysis indicated that the transport processes were controlled by proximal sources mixing and diverse mobilization in various periods responding to rainstorms. NO3- and Cl- presented different hysteresis behavior as sourced from agricultural activities. DOC increased on the hydrograph rising limb and was controlled by a transport-limited regime. However, the hysteresis behavior of most weathering products and DIC were regulated by a process-limited regime in the karstic critical zone. Overall, biogeochemical processes, hydrogeological properties, storm intensity/magnitude and the timing of storms (antecedent conditions) are main factors influencing the response of hydrochemical variables and dissolved carbon to storm events.

Wind farm development on peatlands increases fluvial macronutrient loading.

Wind farms can help to mitigate increasing atmospheric carbon (C) emissions. However, disturbance caused by wind farm development must not have lasting deleterious impacts on landscape C sequestration. To understand the effects of wind farm development on peatlands, we monitored streamwater at Europe's second largest onshore wind farm (539 MW), Whitelee, Scotland, for 31 months. Using nested catchment sampling to understand impacts on water quality, increasing macronutrient concentrations and exports were associated with wind farm development, particularly forest-felling and borrow pits. Low/poor water quality occurred in small headwater catchments most disturbed by development. At the site exit, dissolved organic C and soluble reactive phosphorus (SRP) concentrations increased during construction, though [SRP] recovery occurred within 2 years. Since C was lost and streamwater quality negatively affected, we propose future good practice measures for wind farm development, including limiting total disturbance within individual catchments and locating borrow pits, where deemed necessary, off site avoiding peatlands.

Rainfall-driven E. coli transfer to the stream-conduit network observed through increasing spatial scales in mixed land-use paddy farming karst terrain.

Karst aquifers have distinctive hydrology and supply 25% of the world's population with drinking water, making them a critical geological setting for understanding and managing microbial water pollution. Rainfall causes elevated concentrations and loading of faecal microorganisms, e.g. E. coli, in catchment surface and groundwater systems, increasing the risk of human exposure to faecally-contaminated water. However, effective management of microbial water quality in complex karst catchments is constrained by limited understanding of E. coli - discharge responses to rainfall. We analysed how rainfall events of varying magnitude (2.4-100 mm) control E. coli-discharge dynamics at increasing spatial scales in a mixed land-use karst catchment in southwest China. During the wet season, hourly water sampling was undertaken throughout five storm events to characterise in high detail E. coli emergence with resulting flow across multiple sites of varying catchment area, stream order, and land-use. E. coli concentration was found to increase by 1-3 orders of magnitude following rainfall events. Maximum E. coli concentration and speed of E. coli recession were influenced by rainfall (amount, intensity), timing of agricultural activities, and position in the hydrological system. For high intensity events ∼90% of the cumulative E. coli export occurred within 48 h. E. coli concentration increased with increasing discharge at all sites. E. coli concentration at low discharge was higher in the headwaters than at the catchment outlet, while the rate of increase in E. coli concentration with increasing discharge appears to follow the opposite trend, being higher at the catchment outlet than the headwaters. This was attributed to the decreasing flow path gradient and increasing degree of development of the fissure network, but further event monitoring at varying catchment scales is required to confirm this relationship. The results provide novel insight into how rainfall characteristics combine with land-use and catchment hydrology to control E. coli export in karst landscapes.

C mobilisation in disturbed tropical peat swamps: old DOC can fuel the fluvial efflux of old carbon dioxide, but site recovery can occur.

Southeast-Asian peat swamp forests have been significantly logged and converted to plantation. Recently, to mitigate land degradation and C losses, some areas have been left to regenerate. Understanding how such complex land use change affects greenhouse gas emissions is essential for modelling climate feedbacks and supporting land management decisions. We carried out field research in a Malaysian swamp forest and an oil palm plantation to understand how clear-felling, drainage, and illegal and authorized conversion to oil palm impacted the C cycle, and how the C cycle may change if such logging and conversion stopped. We found that both the swamp forest and the plantation emit centuries-old CO2 from their drainage systems in the managed areas, releasing sequestered C to the atmosphere. Oil palm plantations are an iconic symbol of tropical peatland degradation, but CO2 efflux from the recently-burnt, cleared swamp forest was as old as from the oil palm plantation. However, in the swamp forest site, where logging had ceased approximately 30 years ago, the age of the CO2 efflux was modern, indicating recovery of the system can occur. 14C dating of the C pool acted as a tracer of recovery as well as degradation and offers a new tool to assess efficacy of restoration management. Methane was present in many sites, and in higher concentrations in slow-flowing anoxic systems as degassing mechanisms are not strong. Methane loading in freshwaters is rarely considered, but this may be an important C pool in restored drainage channels and should be considered in C budgets and losses.

Average daily flow of microplastics through a tertiary wastewater treatment plant over a ten-month period.

Microplastics (MPs, <5 mm in size) are classified as emerging contaminants but treatment processes are not designed to remove these small particles. Wastewater treatment systems have been proposed as pathways for MPs pollution to receiving waters but quantitative and qualitative data on MP occurrence and transport remains limited, hindering risk assessment and regulation. Here, for the first time, the stepwise abundance and loading of MPs (60-2800 µm) in a tertiary wastewater treatment plant in the UK was assessed by sampling from May 2017 to February 2018. Microplastics were found in all sampling campaigns, with an average inflow of 8.1 × 108 (95% CI, 3.8 × 108 to 1.2 × 109) items day-1. Their prevalence decreased from influent to final effluent. Overall abundances decreased on average by 6%, 68%, 92%, and 96% after the pre-treatment, primary, secondary, and tertiary treatment stages respectively, although considerable variability occurred throughout the year. Sufficient particles remained in the treated effluent to generate an average discharge of 2.2 × 107 (95% CI, 1.2 × 107 to 3.2 × 107]) particles day-1 to the recipient river. Secondary MPs were predominant, while primary MP abundances were minimal. Fibres comprised 67% of all items, followed by films (18%) and fragments (15%). Chemical characterisation confirmed the presence of different types of polymers, with polypropylene fibres and fragments most abundant (23%). This research informs understanding of how wastewater effluent may channel MPs to the natural environment and their composition, and helps understand control points for optimising advanced treatment processes.

Microscopy and elemental analysis characterisation of microplastics in sediment of a freshwater urban river in Scotland, UK.

Understanding of the sources, fate, and impact of microplastics (MPs, < 5 mm) remains limited, particularly in freshwater environments, while limited comparability across available surveys hinders adequate monitoring and risk assessment of these contaminants. Here, the distribution of microscopic debris in an urban river close to the marine environment in the West of Scotland was investigated to assess concentration and distribution of primary and secondary MPs. Also, the efficiency of light and scanning electron microscopy with energy-dispersive spectroscopy (SEM-EDS) was evaluated for characterisation and quantification of MPs sized 2.8 mm-11 µm. Bank sediment samples were collected twice from the River Kelvin in Glasgow and were size-fractionated and processed for extraction of MPs by density separation. Sample MPs spiking and use of procedural blanks allowed the influence of processing on field data quality to be considered. Total abundances were 161-432 MPs kg-1 dry sediment, with fibres as the dominant type, comprising > 88% of total counts. Nevertheless, fibres in blanks suggest potential contributions from atmospheric contamination. Moreover, fibres concentrated mainly in fractions < 0.09 mm suggesting that their fate may be influenced by drivers of fine sediment dynamics in rivers. While no primary MPs were observed, metallic and glass pellets were present in high abundances in settled material and could be easily misidentified by visual inspection, demonstrating that compositional analysis is needed to avoid analytical errors from MP misidentification and overestimation. SEM-EDS allowed for a quick screening of plastic vs non-plastic pellets and improved identification of smaller fragments, whereas more advanced techniques are needed for proper identification of fibres. This study is the first to report on MPs in freshwater rivers in Scotland and suggests that diffuse sources of pollution may be delivering secondary MPs to the river. Their sources, fate, and risk in these systems will thus warrant further attention.

Increased fire severity alters initial vegetation regeneration across Calluna-dominated ecosystems.

Calluna vulgaris-dominated habitats are valued for ecosystem services such as carbon storage and for their conservation importance. Climate and environmental change are altering their fire regimes. In particular, more frequent summer droughts will result in higher severity wildfires. This could alter the plant community composition of Calluna habitats and thereby influence ecosystem function. To study the effect of fire severity on community composition we used rain-out shelters to simulate drought prior to experimental burns at two Calluna-dominated sites, a raised bog and a heathland. We analysed species abundance in plots surveyed ca. 16 months after fire in relation to burn severity (indicated by fire-induced soil heating). We found that fire severity was an important control on community composition at both sites. Higher fire severity increased the abundance of ericoids, graminoids and acrocarpous mosses, and decreased the abundance of pleurocarpous mosses compared to lower severity fires. At the raised bog, the keystone species Sphagnum capillifolium and Eriophorum vaginatum showed no difference in regeneration with fire severity. Species and plant functional type beta-diversity increased following fire, and was similar in higher compared to lower severity burns. Our results further our understanding of the response of Calluna-dominated habitats to projected changes in fire regimes, and can assist land managers using prescribed fires in selecting burning conditions to achieve management objectives.

How can we improve understanding of faecal indicator dynamics in karst systems under changing climatic, population, and land use stressors? - Research opportunities in SW China.

Human exposure to water contaminated with faeces is a leading cause of worldwide ill-health. Contaminated water can be transmitted rapidly in karst terrain as a result of the connectivity of surface and groundwater systems, high transmissivity of aquifers over large areas, and well-developed underground conduit systems. Faecal indicator organisms (FIOs) are the most widely-used indicator of faecal contamination and microbial water quality; however, the conceptualisation of FIO risk and associated sources, pathways, and survival dynamics of FIOs in karst landscapes requires a degree of modification from traditional conceptual models of FIO fate and transfer in non-karst systems. While a number of reviews have provided detailed accounts of the state-of-the-science concerning FIO dynamics in catchments, specific reference to the uniqueness of karst and its influence on FIO fate and transfer is a common omission. In response, we use a mixed methods approach of critical review combined with a quantitative survey of 372 residents of a typical karst catchment in the southwest China karst region (SWCKR) to identify emerging research needs in an area where much of the population lives in poverty and is groundwater dependent. We found that the key research needs are to understand: 1) overland and subsurface FIO export pathways in karst hydrology under varying flow conditions; 2) urban and agricultural sources and loading in mixed land-use paddy farming catchments; 3) FIO survival in paddy farming systems and environmental matrices in karst terrain; 4) sediment-FIO interactions and legacy risk in karst terrain; and 5) key needs for improved hydrological modelling and risk assessment in karst landscapes. Improved knowledge of these research themes will enable the development of evidence-based faecal contamination mitigation strategies for managing land and water resources in the SWCKR, which is highly vulnerable to climate change impacts on water supply and quality of water resources.

Burning increases post-fire carbon emissions in a heathland and a raised bog, but experimental manipulation of fire severity has no effect.

Large amounts of carbon are stored in northern peatlands. There is concern that greater wildfire severity following projected increases in summer drought will lead to higher post-fire carbon losses. We measured soil carbon dynamics in a Calluna heathland and a raised peat bog after experimentally manipulating fire severity. A gradient of fire severity was achieved by simulating drought in 2 × 2 m plots. Ecosystem respiration (ER), net ecosystem exchange (NEE), methane (CH4) flux and concentration of dissolved organic carbon ([DOC], measured at the raised bog only) were measured for up to two years after burning. The response of these carbon fluxes to increased fire severity in drought plots was similar to plots burnt under ambient conditions associated with traditional managed burning. Averaged across all burnt plots, burning altered mean NEE from a net carbon sink at the heathland (-0.33 µmol CO2 m-2 s-1 in unburnt plots) to a carbon source (0.50 µmol m-2 s-1 in burnt plots) and at the raised bog (-0.38 and 0.16 µmol m-2 s-1, respectively). Burning also increased CH4 flux at the raised bog (from 1.16 to 25.3 nmol m-2 s-1 in the summer, when it accounted for 79% of the CO2-equivalent emission). Burning had no significant effect on soil water [DOC].

Fluvial dissolved organic carbon composition varies spatially and seasonally in a small catchment draining a wind farm and felled forestry.

Assessing whether land use, from activities such as wind farm construction and tree-felling, impacts on terrestrial C delivery to rivers has focused on quantifying the loss of dissolved organic carbon (DOC), and not the composition changes. Here we explore how land use influences DOC composition by considering fluvial DOC concentration, [DOC], and spectrophotometric composition of a river draining a peat-rich catchment. We find that in this 5.7km2 catchment differences occur in both the concentration and composition of the DOC in its sub-catchments. This is attributed to differences in how land was used: one tributary (D-WF) drains an area with wind farm construction and forestry in the headwaters, and one tributary (D-FF) drains an area with felled plantation trees. Generally, [DOC] in both streams showed similar seasonal variation, and autumn maxima. However, the felled catchment had greater mean [DOC] than the wind farm catchment. The SUVA254 and E4/E6 indicated DOC in both streams had similar aromaticity and fulvic:humic acid for most of the time, but SUVA410 and E2/E4 indicated less DOC humification in the felled catchment. This may be due to young DOC from the breakdown of residual branches and roots, or more humification in soils in the wind farm area. During the dry months, DOC composition showed more spatial variation: the D-WF DOC had smaller SUVA254 (less total aromatic material) and SUVA410 (fewer humic substances). The decreased E2/E4 in both streams indicated the total aromatic carbon decreased more than humic substances content. Moreover, the larger E4/E6 for D-WF in summer indicated that the humic substances were richer in fulvic acids than humic acids. Soil disturbance associated with forestry-felling likely contributed to the higher [DOC] and release of less-humified material in D-FF. This research indicates drivers of different DOC concentration and composition can exist even in small catchments.

Fire severity is more sensitive to low fuel moisture content on Calluna heathlands than on peat bogs.

Moorland habitats dominated by the dwarf shrub Calluna vulgaris provide important ecosystem services. Drought is projected to intensify throughout their range, potentially leading to increased fire severity as moisture is a key control on severity. We studied the effect of low fuel moisture content (FMC) on fire severity by using 2×2m rain-out shelters prior to completing 19 experimental fires in two sites in Scotland (UK): a dry heath with thin organic soils and a raised bog with deep, saturated peat, both dominated by Calluna vulgaris. Reduced FMC of the moss and litter (M/L) layer at both sites, and the soil moisture of the dry heath, increased fire-induced consumption of the M/L layer and soil heating at both sites. Increase in fire severity was greater at the dry heath than at the raised bog, e.g. average maximum temperatures at the soil surface increased from 31°C to 189°C at the dry heath, but only from 10°C to 15°C at the raised bog. Substantial M/L layer consumption was observed when its FMC was below 150%. This led to larger seasonal and daily soil temperature fluctuation, particularly at the dry heath during warm months. The results suggest that low FMC following predicted changes in climate are likely to increase wildfire severity and that the impact on vegetation composition and carbon stores may be greater at heathlands than at peatlands. Managed burning aiming to minimise fire severity (e.g. ignition of the M/L layer and exposure to lethal temperatures of ericoid seeds) should be carried out when the FMC of the M/L layer is above 150% and the FMC of the soil is above 200-300%.

Leaving moss and litter layers undisturbed reduces the short-term environmental consequences of heathland managed burns.

Variation in the structure of ground fuels, i.e. the moss and litter (M/L) layer, may be an important control on fire severity in heather moorlands and thus influence vegetation regeneration and soil carbon dynamics. We completed experimental fires in a Calluna vulgaris-dominated heathland to study the role of the M/L layer in determining (i) fire-induced temperature pulses into the soil and (ii) post-fire soil thermal dynamics. Manually removing the M/L layer before burning increased fire-induced soil heating, both at the soil surface and 2 cm below. Burnt plots where the M/L layer was removed simulated the fuel structure after high severity fires where ground fuels are consumed but the soil does not ignite. Where the M/L layer was manually removed, either before or after the fire, post-fire soil thermal dynamics showed larger diurnal and seasonal variation, as well as similar patterns to those observed after wildfires, compared to burnt plots where the M/L layer was not manipulated. We used soil temperatures to explore potential changes in post-fire soil respiration. Simulated high fire severity (where the M/L layer was manually removed) increased estimates of soil respiration in warm months. With projected fire regimes shifting towards higher severity fires, our results can help land managers develop strategies to balance ecosystem services in Calluna-dominated habitats.

A novel modified-indirect ELISA based on spherical body protein 4 for detecting antibody during acute and long-term infections with diverse Babesia bovis strains.

BACKGROUND: Cattle persistently infected with Babesia bovis are reservoirs for intra- and inter-herd transmission. Since B. bovis is considered a persistent infection, developing a reliable, high-throughput assay that detects antibody during all stages of the infection could be pivotal for establishing better control protocols. METHODS: A modified indirect enzyme-linked immunosorbent assay (MI-ELISA) was developed using the spherical body protein-4 (SBP4) of B. bovis to detect antibody against diverse strains through all infection stages in cattle. This SBP4 MI-ELISA was evaluated for sensitivity and specificity against field sera from regions with endemic and non-endemic B. bovis. Sera were also evaluated from cattle infected experimentally with various doses and strains during acute and persistent infection with parasitemia defined by nested PCR. RESULTS: The format variables for SBP4 MI-ELISA were optimized and the cutoff for positive and negative interpretation was determined based on receiver operating characteristic curve analysis using B. bovis positive and negative sera tested in the reference immunofluorescence assay (IFA). The diagnostic specificity of the SBP4 MI-ELISA using IFA-negative sera collected from Texas was 100%, significantly higher than the cELISA (90.4%) based on an epitope in the rhoptry-associated protein-1 (RAP-1 cELISA). The diagnostic sensitivity of the SBP4 MI-ELISA was 98.7% using the IFA-positive sera collected from several areas of Mexico, in contrast to that of the RAP-1 cELISA at 60% using these same sera. In cattle infected with low and high doses of three B. bovis strains, the SBP4 MI-ELISA remained antibody positive for 11 months or more after initial detection at 10 to 13 days post-inoculation. However, the RAP-1 cELISA did not reliably detect antibody after eight months post-inoculation despite the fact that parasitemia was occasionally detectable by PCR. Furthermore, initial antibody detection by RAP-1 cELISA in low-dose infected animals was delayed approximately nine and a half days compared to the SBP4 MI-ELISA. CONCLUSIONS: These results demonstrate excellent diagnostic sensitivity and specificity of the novel SBP4 MI-ELISA for cattle with acute and long-term carrier infections. It is posited that use of this assay in countries that have B. bovis-endemic herds may be pivotal in preventing the spread of this disease to non-endemic herds.

Diel Surface Temperature Range Scales with Lake Size.

Ecological and biogeochemical processes in lakes are strongly dependent upon water temperature. Long-term surface warming of many lakes is unequivocal, but little is known about the comparative magnitude of temperature variation at diel timescales, due to a lack of appropriately resolved data. Here we quantify the pattern and magnitude of diel temperature variability of surface waters using high-frequency data from 100 lakes. We show that the near-surface diel temperature range can be substantial in summer relative to long-term change and, for lakes smaller than 3 km2, increases sharply and predictably with decreasing lake area. Most small lakes included in this study experience average summer diel ranges in their near-surface temperatures of between 4 and 7°C. Large diel temperature fluctuations in the majority of lakes undoubtedly influence their structure, function and role in biogeochemical cycles, but the full implications remain largely unexplored.