The ecosystem wilting point defines drought response and recovery of a Quercus-Carya forest.

Soil and atmospheric droughts increasingly threaten plant survival and productivity around the world. Yet, conceptual gaps constrain our ability to predict ecosystem-scale drought impacts under climate change. Here, we introduce the ecosystem wilting point (ΨEWP ), a property that integrates the drought response of an ecosystem's plant community across the soil-plant-atmosphere continuum. Specifically, ΨEWP defines a threshold below which the capacity of the root system to extract soil water and the ability of the leaves to maintain stomatal function are strongly diminished. We combined ecosystem flux and leaf water potential measurements to derive the ΨEWP of a Quercus-Carya forest from an "ecosystem pressure-volume (PV) curve," which is analogous to the tissue-level technique. When community predawn leaf water potential (Ψpd ) was above ΨEWP (=-2.0 MPa), the forest was highly responsive to environmental dynamics. When Ψpd fell below ΨEWP , the forest became insensitive to environmental variation and was a net source of carbon dioxide for nearly 2 months. Thus, ΨEWP is a threshold defining marked shifts in ecosystem functional state. Though there was rainfall-induced recovery of ecosystem gas exchange following soaking rains, a legacy of structural and physiological damage inhibited canopy photosynthetic capacity. Although over 16 growing seasons, only 10% of Ψpd observations fell below ΨEWP , the forest is commonly only 2-4 weeks of intense drought away from reaching ΨEWP , and thus highly reliant on frequent rainfall to replenish the soil water supply. We propose, based on a bottom-up analysis of root density profiles and soil moisture characteristic curves, that soil water acquisition capacity is the major determinant of ΨEWP , and species in an ecosystem require compatible leaf-level traits such as turgor loss point so that leaf wilting is coordinated with the inability to extract further water from the soil.

Confronting the water potential information gap.

Water potential directly controls the function of leaves, roots, and microbes, and gradients in water potential drive water flows throughout the soil-plant-atmosphere continuum. Notwithstanding its clear relevance for many ecosystem processes, soil water potential is rarely measured in-situ, and plant water potential observations are generally discrete, sparse, and not yet aggregated into accessible databases. These gaps limit our conceptual understanding of biophysical responses to moisture stress and inject large uncertainty into hydrologic and land surface models. Here, we outline the conceptual and predictive gains that could be made with more continuous and discoverable observations of water potential in soils and plants. We discuss improvements to sensor technologies that facilitate in situ characterization of water potential, as well as strategies for building new networks that aggregate water potential data across sites. We end by highlighting novel opportunities for linking more representative site-level observations of water potential to remotely-sensed proxies. Together, these considerations offer a roadmap for clearer links between ecohydrological processes and the water potential gradients that have the 'potential' to substantially reduce conceptual and modeling uncertainties.

The xylem of anisohydric Quercus alba L. is more vulnerable to embolism than isohydric codominants.

The coordination of plant leaf water potential (ΨL ) regulation and xylem vulnerability to embolism is fundamental for understanding the tradeoffs between carbon uptake and risk of hydraulic damage. There is a general consensus that trees with vulnerable xylem more conservatively regulate ΨL than plants with resistant xylem. We evaluated if this paradigm applied to three important eastern US temperate tree species, Quercus alba L., Acer saccharum Marsh. and Liriodendron tulipifera L., by synthesizing 1600 ΨL observations, 122 xylem embolism curves and xylem anatomical measurements across 10 forests spanning pronounced hydroclimatological gradients and ages. We found that, unexpectedly, the species with the most vulnerable xylem (Q. alba) regulated ΨL less strictly than the other species. This relationship was found across all sites, such that coordination among traits was largely unaffected by climate and stand age. Quercus species are perceived to be among the most drought tolerant temperate US forest species; however, our results suggest their relatively loose ΨL regulation in response to hydrologic stress occurs with a substantial hydraulic cost that may expose them to novel risks in a more drought-prone future.

Mild water and salt stress improve water use efficiency by decreasing stomatal conductance via osmotic adjustment in field maize.

Water and salt stress often occur simultaneously in heavily irrigated arid agricultural areas, yet they are usually studied in isolation. To understand the physiological bases of water use efficiency (WUE) of field-grown maize (Zea mays) at multi-scales under combined water and salt stress, we investigated the joint effects of water and salt stress on physiology, growth, yield, and WUE of two genotypes (XY335 and ZD958). We measured leaf stomatal conductance (gs), net photosynthesis rate (A) and hydraulic traits, whole-plant growth and water use (ET), and final biomass and grain yield. Leaf osmotic adjustment was a key trait of the physiological differences between XY335 and ZD958 under water and salt stress. Although the responses of the two genotypes were different, mild water and salt stress improved intrinsic water use efficiency (iWUE = A/gs) by (i) decreasing gsvia increasing osmotic adjustment and hydraulic resistance, and (ii) declining A via increasing stomatal limitations rather than reducing photosynthetic capacity. Joint water and salt stress had a synergistic effect on reproductive growth and grain formation of maize. Mild water and salt stress reduced ET, stabilized grain yield, and improved grain WUE via declining gs, maintaining photosynthetic capacity, and improving harvest index. Collectively, our study provides a novel insight into the physiological mechanisms of WUE and demonstrates an approach for the efficient management of water and salt by using a growth stage-based deficit irrigation strategy or/and selecting genotypes with strong osmotic adjustment capacity and high harvest index.

Detecting forest response to droughts with global observations of vegetation water content.

Droughts in a warming climate have become more common and more extreme, making understanding forest responses to water stress increasingly pressing. Analysis of water stress in trees has long focused on water potential in xylem and leaves, which influences stomatal closure and water flow through the soil-plant-atmosphere continuum. At the same time, changes of vegetation water content (VWC) are linked to a range of tree responses, including fluxes of water and carbon, mortality, flammability, and more. Unlike water potential, which requires demanding in situ measurements, VWC can be retrieved from remote sensing measurements, particularly at microwave frequencies using radar and radiometry. Here, we highlight key frontiers through which VWC has the potential to significantly increase our understanding of forest responses to water stress. To validate remote sensing observations of VWC at landscape scale and to better relate them to data assimilation model parameters, we introduce an ecosystem-scale analog of the pressure-volume curve, the non-linear relationship between average leaf or branch water potential and water content commonly used in plant hydraulics. The sources of variability in these ecosystem-scale pressure-volume curves and their relationship to forest response to water stress are discussed. We further show to what extent diel, seasonal, and decadal dynamics of VWC reflect variations in different processes relating the tree response to water stress. VWC can also be used for inferring belowground conditions-which are difficult to impossible to observe directly. Lastly, we discuss how a dedicated geostationary spaceborne observational system for VWC, when combined with existing datasets, can capture diel and seasonal water dynamics to advance the science and applications of global forest vulnerability to future droughts.

Eastern US deciduous tree species respond dissimilarly to declining soil moisture but similarly to rising evaporative demand.

Hydraulic stress in plants occurs under conditions of low water availability (soil moisture; θ) and/or high atmospheric demand for water (vapor pressure deficit; D). Different species are adapted to respond to hydraulic stress by functioning along a continuum where, on one hand, they close stomata to maintain a constant leaf water potential (ΨL) (isohydric species), and on the other hand, they allow ΨL to decline (anisohydric species). Differences in water-use along this continuum are most notable during hydrologic stress, often characterized by low θ and high D; however, θ and D are often, but not necessarily, coupled at time scales of weeks or longer, and uncertainty remains about the sensitivity of different water-use strategies to these variables. We quantified the effects of both θ and D on canopy conductance (Gc) among widely distributed canopy-dominant species along the isohydric-anisohydric spectrum growing along a hydroclimatological gradient. Tree-level Gc was estimated using hourly sap flow observations from three sites in the eastern United States: a mesic forest in western North Carolina and two xeric forests in southern Indiana and Missouri. Each site experienced at least 1 year of substantial drought conditions. Our results suggest that sensitivity of Gc to θ varies across sites and species, with Gc sensitivity being greater in dry than in wet sites, and greater for isohydric compared with anisohydric species. However, once θ limitations are accounted for, sensitivity of Gc to D remains relatively constant across sites and species. While D limitations to Gc were similar across sites and species, ranging from 16 to 34% reductions, θ limitations to Gc ranged from 0 to 40%. The similarity in species sensitivity to D is encouraging from a modeling perspective, though it implies that substantial reduction to Gc will be experienced by all species in a future characterized by higher D.

Does overwintering change the inoculum effect on methane emissions from stored liquid manure?

Greenhouse gas (GHG) emissions, especially methane (CH4 ), from manure storage facilities can be substantial. Methane production requires adapted microbial communities ("inoculum") to be present in the manure. Complete removal of liquid dairy manure (thus removing all inoculum) from storage tanks in the spring has been shown to significantly reduce CH4 emissions over the following warm season. This study examined whether the same mitigation effect would occur after fall removal of liquid dairy manure. Emissions of CH4 , nitrous oxide (N2 O), ammonia (NH3 ), and CO2 were measured from six 11.88-m3 tanks equipped with flow-through chambers. There were three inoculated controls (20% inoculum) and three uninoculated treatments, where inoculum was completely removed in the fall/winter (0% inoculum). Direct N2 O and NH3 (indirect N2 O) were minor contributors to the total GHG budget, contributing <2% on a CO2 equivalent (CO2 e) basis. Removal of inoculum led to a 34% decrease in total emissions on a CO2 e basis and to a 29% decrease in the CH4 conversion factor compared with the inoculated control (0.37 vs. 0.52; p = .01). Overall, removing inoculum in the fall reduced CH4 emissions from manure storage tanks; however, fall inoculum removal was less effective than in a previous study where inoculum was removed in the spring. The timing of inoculum removal may affect the efficiency of this CH4 mitigation strategy. However, this method may be impractical for larger manure storage tanks. Further study is required to overcome challenges of time-sensitive, complete inoculum removal from farm-scale storage tanks.

Top-Down Constraints on Methane Point Source Emissions From Animal Agriculture and Waste Based on New Airborne Measurements in the U.S. Upper Midwest.

Agriculture and waste are thought to account for half or more of the U.S. anthropogenic methane source. However, current bottom-up inventories contain inherent uncertainties from extrapolating limited in situ measurements to larger scales. Here, we employ new airborne methane measurements over the U.S. Corn Belt and Upper Midwest, among the most intensive agricultural regions in the world, to quantify emissions from an array of key agriculture and waste point sources. Nine of the largest concentrated animal feeding operations in the region and two sugar processing plants were measured, with multiple revisits during summer (August 2017), winter (January 2018), and spring (May-June 2018). We compare the top-down fluxes with state-of-science bottom-up estimates informed by U.S. Environmental Protection Agency methodology and site-level animal population and management practices. Top-down point source emissions are consistent with bottom-up estimates for beef concentrated animal feeding operations but moderately lower for dairies (by 37% on average) and significantly lower for sugar plants (by 80% on average). Swine facility results are more variable. The assumed bottom-up seasonality for manure methane emissions is not apparent in the aircraft measurements, which may be due to on-site management factors that are difficult to capture accurately in national-scale inventories. If not properly accounted for, such seasonal disparities could lead to source misattribution in top-down assessments of methane fluxes.

Sun-induced Chl fluorescence and its importance for biophysical modeling of photosynthesis based on light reactions.

Recent progress in observing sun-induced Chl fluorescence (SIF) provides an unprecedented opportunity to advance photosynthesis research in natural environments. However, we still lack an analytical framework to guide SIF studies and integration with the well-developed active fluorescence approaches. Here, we derive a set of coupled fundamental equations to describe the dynamics of SIF and its relationship with C3 and C4 photosynthesis. These equations show that, although SIF is dynamically as complex as photosynthesis, the measured SIF simplifies photosynthetic modeling from the perspective of light reactions by integrating over the dynamic complexities of photosynthesis. Specifically, the measured SIF contains direct information about the actual electron transport from photosystem II to photosystem I, giving a quantifiable link between light and dark reactions. With much-reduced requirements on inputs and parameters, the light-reactions-centric, SIF-based biophysical model complements the traditional, dark-reactions-centric biochemical model of photosynthesis. The SIF-photosynthesis relationship, however, is nonlinear. This is because photosynthesis saturates at high light whereas SIF has a stronger tendency to keep increasing, as fluorescence quantum yield has a relatively muted sensitivity to light levels. Successful applications of the SIF-based model of photosynthesis will depend on a predictive understanding of several previously underexplored physiological and biophysical processes. Advances can be facilitated by coordinated efforts in plant physiology, remote sensing, and eddy covariance flux observations.

Error characterization of methane fluxes and budgets derived from a long-term comparison of open- and closed-path eddy covariance systems.

Wetlands represent the dominant natural source of methane (CH4) to the atmosphere. Thus, substantial effort has been spent examining the CH4 budgets of global wetlands via continuous ecosystem-scale measurements using the eddy covariance (EC) technique. Robust error characterization for such measurements, however, remains a major challenge. Here, we quantify systematic, random and gap-filling errors and the resulting uncertainty in CH4 fluxes using a 3.5 year time series of simultaneous open- and closed path CH4 flux measurements over a sub-boreal wetland. After correcting for high- and low frequency flux attenuation, the magnitude of systematic frequency response errors were negligible relative to other uncertainties. Based on three different random flux error estimations, we found that errors of the CH4 flux measurement systems were smaller in magnitude than errors associated with the turbulent transport and flux footprint heterogeneity. Errors on individual half-hourly CH4 fluxes were typically 6%-41%, but not normally distributed (leptokurtic), and thus need to be appropriately characterized when fluxes are compared to chamber-derived or modeled CH4 fluxes. Integrated annual fluxes were only moderately sensitive to gap-filling, based on an evaluation of 4 different methods. Calculated budgets agreed on average to within 7% (≤ 1.5 g - CH4 m-2 yr-1). Marginal distribution sampling using open source code was among the best-performing of all the evaluated gap-filling approaches and it is therefore recommended given its transparency and reproducibility. Overall, estimates of annual CH4 emissions for both EC systems were in excellent agreement (within 0.6 g - CH4 m-2 yr-1) and averaged 18 g - CH4 m-2 yr-1. Total uncertainties on the annual fluxes were larger than the uncertainty of the flux measurement systems and estimated between 7-17%. Identifying trends and differences among sites or site years requires that the observed variability exceeds these uncertainties.

Source Partitioning of Methane Emissions and its Seasonality in the U.S. Midwest.

The methane (CH4) budget and its source partitioning are poorly constrained in the Midwestern United States. We used tall tower (185 m) aerodynamic flux measurements and atmospheric scale factor Bayesian inversions to constrain the monthly budget and to partition the total budget into natural (e.g., wetlands) and anthropogenic (e.g., livestock, waste, and natural gas) sources for the period June 2016 to September 2017. Aerodynamic flux observations indicated that the landscape was a CH4 source with a mean annual CH4 flux of +13.7 ± 0.34 nmol m-2 s-1 and was rarely a net sink. The scale factor Bayesian inversion analyses revealed a mean annual source of +12.3 ± 2.1 nmol m-2 s-1. Flux partitioning revealed that the anthropogenic source (7.8 ± 1.6 Tg CH4 yr-1) was 1.5 times greater than the bottom-up gridded United States Environmental Protection Agency inventory, in which livestock and oil/gas sources were underestimated by 1.8-fold and 1.3-fold, respectively. Wetland emissions (4.0 ± 1.2 Tg CH4 yr-1) were the second largest source, accounting for 34% of the total budget. The temporal variability of total CH4 emissions was dominated by wetlands with peak emissions occurring in August. In contrast, emissions from oil/gas and other anthropogenic sources showed relatively weak seasonality.

Nitrous oxide emissions are enhanced in a warmer and wetter world.

Nitrous oxide (N2O) has a global warming potential that is 300 times that of carbon dioxide on a 100-y timescale, and is of major importance for stratospheric ozone depletion. The climate sensitivity of N2O emissions is poorly known, which makes it difficult to project how changing fertilizer use and climate will impact radiative forcing and the ozone layer. Analysis of 6 y of hourly N2O mixing ratios from a very tall tower within the US Corn Belt-one of the most intensive agricultural regions of the world-combined with inverse modeling, shows large interannual variability in N2O emissions (316 Gg N2O-N⋅y-1 to 585 Gg N2O-N⋅y-1). This implies that the regional emission factor is highly sensitive to climate. In the warmest year and spring (2012) of the observational period, the emission factor was 7.5%, nearly double that of previous reports. Indirect emissions associated with runoff and leaching dominated the interannual variability of total emissions. Under current trends in climate and anthropogenic N use, we project a strong positive feedback to warmer and wetter conditions and unabated growth of regional N2O emissions that will exceed 600 Gg N2O-N⋅y-1, on average, by 2050. This increasing emission trend in the US Corn Belt may represent a harbinger of intensifying N2O emissions from other agricultural regions. Such feedbacks will pose a major challenge to the Paris Agreement, which requires large N2O emission mitigation efforts to achieve its goals.

Greenhouse Gas Emissions from Stored Dairy Slurry from Multiple Farms.

A significant need exists to improve our understanding of the extent of greenhouse gas emissions from the storage of livestock manure to both improve the reliability of inventory assessments and the impact of beneficial management practice adoption. Factors affecting the extent and variability of greenhouse gas emissions from stored dairy manure were investigated. Emissions from six slurries stored in clean concrete tanks under identical "warm-season" conditions were monitored consecutively over 173 d (18°C average air temperature). Methane (CH) emissions varied considerably among the manures from 6.3 to 25.9 g m d and accounted for ∼96% of the total CO equivalent greenhouse gas emissions. The duration of the lag period, when methane emissions were near baseline levels, varied from 30 to 90 d from the beginning of storage. As a result, CH emissions were poorly correlated with air temperature prior to the time of peak emissions (i.e., the initial 48 to 108 d of storage) but improved afterward. The air temperature following the time of the peak CH flux and the length of the active methanogenesis period (i.e., when the daily CH emissions ≥ 7.6 g m d) were highly correlated with CH emissions ( = 0.98, < 0.01). Methane conversion factors (MCFs) ranged from 0.08 to 0.52 for the different manures. The MCFs generated from existing CH emission models were correlated ( = 0.68, = 0.02) to MCFs calculated for the active methanogenesis period for manure containing wood bedding. A temperature component was added that improved the accuracy ( = 0.82, < 0.01). This demonstrated that an improved understanding of lag period dynamics will enhance stored dairy manure greenhouse gas emission inventory calculations.

Performance of Seasonally and Continuously Loaded Constructed Wetlands Treating Dairy Farm Wastewater.

A 2-yr study compared the performance of seasonally and continuously loaded constructed wetlands treating dairy farm wastewater. One wetland was loaded during the growing season (GS) periods only, while the other was continuously loaded. Weekly samples were analyzed for 5-d biochemical oxygen demand (BOD), total suspended solids (TSS), total Kjeldahl N (TKN), total ammoniacal N (TAN), total P (TP), and . Annual average daily mass removal rates (kg ha) were similar for both wetlands in both years; however, seasonal differences were observed. With the exception of BOD in Year 2, average daily GS areal mass removal rates were higher for the seasonal wetland. However, GS mass exports from the seasonal wetland were higher by 28 to 94%, with the exception of BOD in Year 1. Annual mass reductions (MRs; %) for nutrients were higher for the continuous wetland in both years. Annual MRs were similar for in both years and for TSS in Year 2. Annual mass exports from the seasonal wetland were higher for nutrients and by 14 to 77% in both years. Pollutant MRs generally decreased during the nongrowing season (NGS) for the continuous wetland; however, in Year 2 when lower loading rates were used, the wetland still removed 84 to 99% of the pollutant masses. The continuous wetland also performed better during periods of high flow that occurred during the GS. Although there were minimal differences in annual treatment performance, continuously loaded systems require less additional infrastructure and should require less maintenance and may, therefore, be more attractive for agricultural applications.

Indirect nitrous oxide emissions from streams within the US Corn Belt scale with stream order.

N2O is an important greenhouse gas and the primary stratospheric ozone depleting substance. Its deleterious effects on the environment have prompted appeals to regulate emissions from agriculture, which represents the primary anthropogenic source in the global N2O budget. Successful implementation of mitigation strategies requires robust bottom-up inventories that are based on emission factors (EFs), simulation models, or a combination of the two. Top-down emission estimates, based on tall-tower and aircraft observations, indicate that bottom-up inventories severely underestimate regional and continental scale N2O emissions, implying that EFs may be biased low. Here, we measured N2O emissions from streams within the US Corn Belt using a chamber-based approach and analyzed the data as a function of Strahler stream order (S). N2O fluxes from headwater streams often exceeded 29 nmol N2O-N m(-2) ⋅ s(-1) and decreased exponentially as a function of S. This relation was used to scale up riverine emissions and to assess the differences between bottom-up and top-down emission inventories at the local to regional scale. We found that the Intergovernmental Panel on Climate Change (IPCC) indirect EF for rivers (EF5r) is underestimated up to ninefold in southern Minnesota, which translates to a total tier 1 agricultural underestimation of N2O emissions by 40%. We show that accounting for zero-order streams as potential N2O hotspots can more than double the agricultural budget. Applying the same analysis to the US Corn Belt demonstrates that the IPCC EF5r underestimation explains the large differences observed between top-down and bottom-up emission estimates.

Δ(9) desaturase protein expression and fatty acid composition of longissimus dorsi muscle in lambs fed green herbage or concentrate with or without added tannins.

The aims of this study were to investigate the effect of feeding system and of supplementation of tannins (8.93% DM) on the relationship between intramuscular fat content, fatty acid composition and Δ(9)desaturase (Δ(9)d) protein expression in longissimus dorsi muscle of lamb. Twenty-eight Comisana lambs (age 45days) were fed either vetch (Vicia sativa) or concentrate. The herbage diet was (i) lower in saturated fatty acids (especially in C16:0), C18:1 n-9 and in C18:2 n-6; (ii) higher in C16:1 and C18:3 n-3 when compared to concentrate. Within each feeding system the lambs were divided into two sub-groups, one of which received the diet without tannins supplementation, and the other was fed the diets supplemented with the tannins from Quebracho (Schinopsis lorentzii). The animals were slaughtered at age 105days. The concentrate feeding system increased (p<0.01) the total intramuscular fat content and the amount of SFA, MUFA and n-6 PUFA and decreased the level of n-3 PUFA (p=0.05) when compared to the vetch-fed animals but did not affect Δ(9) desaturase protein expression. There was no correlation between Δ(9)d protein expression and total intramuscular fatty acids, CLA and MUFA level. It was suggested that in ruminants, in contrast to monogastric animals, Δ(9)d expression does not play the key role in intramuscular fatty acids formation. Tannins supplementation resulted in higher (p<0.05) muscle levels of transC18:1 and C18:2 n-6. It has also increased Δ(9)d expression in the case of herbage-based diet but not in the case of concentrate-based diet. The mechanism of tannins action on the enzyme expression needs to be elucidated.

Regulation of CYP2A6 protein expression by skatole, indole, and testicular steroids in primary cultured pig hepatocytes.

CYP2A6 is one of the enzymes involved in the hepatic metabolism of a naturally produced compound, skatole, in the pig. Low CYP2A6 activity has been linked to excessive accumulation of skatole in pig adipose tissue and development of the phenomenon "boar taint." CYP2A6 activity varies between male and female animals, suggesting the involvement of sex hormones in regulation of the enzyme activity and/or expression. The present study investigated whether pig hepatic CYP2A6 protein expression is regulated by the testicular steroids testosterone, androstenone, or estrone sulfate using primary cultured hepatocytes as a model system. The study has also examined whether CYP2A6 expression can be modulated by the boar taint compounds skatole and indole. The research has established that androstenone inhibits CYP2A6 protein expression at the concentration of 1, 10, and 100 nM by 55, 37, and 44%, respectively. In contrast to androstenone, skatole and indole (final concentrations, 1, 10, and 100 nM) had a stimulatory effect on CYP2A6 expression. The effect of indole was more pronounced than that of skatole (maximum induction by 145 and 70%, respectively). Estrone sulfate and testosterone did not have a significant effect on CYP2A6 protein level. This is, as far as we know, the first communication to report the regulation of pig hepatic CYP2A6 expression by steroids and boar taint compounds. The hormonal modulation of CYP2A6 expression might contribute to gender-related differences in pig hepatic CYP2A6 activity and skatole accumulation in pig adipose tissue.

Use of a particulate extracellular matrix bioscaffold for treatment of acquired urinary incontinence in dogs.

OBJECTIVE: To evaluate use of a particulate bioscaffold consisting of the extracellular matrix (ECM) of the urinary bladder from pigs for treatment of acquired urinary incontinence in dogs resistant to medical treatment. DESIGN: Case series. ANIMALS: 9 female dogs with acquired urinary incontinence. PROCEDURE: In 6 dogs, 30 mg of particulate ECM in 1.0 mL of a carrier consisting of glycerin and saline 10.9% NaCI) solution was injected into each of 3 equally spaced sites around the circumference of the internal urethral sphincter via an endoscopic technique. In the remaining 3 dogs (control dogs), 1.0 mL of the carrier alone was injected in 3 equally spaced sites around the circumference of the internal urethral sphincter in a similar manner. RESULTS: For dogs treated with the ECM, median duration of urinary continence following treatment was 168 days (range, 84 to 616 days), whereas for the control dogs, median duration of urinary continence following the procedure was 14 days (range, 7 to 31 days). Two of the 3 control dogs were treated with the ECM at the end of the study and were continent for 119 and 252 days. No adverse effects were observed in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that endoscopically guided injection of particulate ECM into the internal urethral sphincter may be useful for the treatment of acquired urinary incontinence in female dogs.