Structure of plant photosystem I in a native assembly state defines PsaF as a regulatory checkpoint.

Plant photosystem I (PSI) consists of at least 13 nuclear-encoded and 4 chloroplast-encoded subunits that together act as a sunlight-driven oxidoreductase. Here we report the structure of a PSI assembly intermediate that we isolated from greening oat seedlings. The assembly intermediate shows an absence of at least eight subunits, including PsaF and LHCI, and lacks photoreduction activity. The data show that PsaF is a regulatory checkpoint that promotes the assembly of LHCI, effectively coupling biogenesis to function.

Investigating the Balance between Structural Conservation and Functional Flexibility in Photosystem I.

Photosynthesis, as the primary source of energy for all life forms, plays a crucial role in maintaining the global balance of energy, entropy, and enthalpy in living organisms. Among its various building blocks, photosystem I (PSI) is responsible for light-driven electron transfer, crucial for generating cellular reducing power. PSI acts as a light-driven plastocyanin-ferredoxin oxidoreductase and is situated in the thylakoid membranes of cyanobacteria and the chloroplasts of eukaryotic photosynthetic organisms. Comprehending the structure and function of the photosynthetic machinery is essential for understanding its mode of action. New insights are offered into the structure and function of PSI and its associated light-harvesting proteins, with a specific focus on the remarkable structural conservation of the core complex and high plasticity of the peripheral light-harvesting complexes.

The importance of consensus science to managing phosphorus in the environment: SERA-17 and the legacy of Andrew Sharpley.

Phosphorus (P) loss from agricultural systems to surface waters, and ultimately, eutrophication, presents a wicked problem requiring transdisciplinary solutions. The mission of SERA-17 (Southern Extension and Research Advisory Information Exchange Group-17) has been to address this problem by developing "Innovative Solutions to Minimize Phosphorus Losses from Agriculture." Over the course of his career, Dr. Andrew Sharpley demonstrated a rare ability to collaboratively achieve consensus around issues related to the science and management of P. The SERA-17 organization served as the central community of experts and stakeholders where that consensus was built and applied. The consensus-based approach, demonstrated by Sharpley and at the core of the SERA-17 organization, was routinely applied to key areas of P science to produce applied outcomes that have been readily adopted: advance foundational science to resolve knowledge gaps and to promote innovation; promote consistency in methods to facilitate comprehensive investigations and conclusions across a diversity of systems; engage diverse stakeholders to prioritize research, and ultimately, ensure that outcomes reflect a plurality of perspectives; and deliver pragmatic solutions that reflect the best information available at a particular time. We review the history of SERA-17 in delivering new science and management recommendations for P, with an eye to elucidating Sharpley's role and legacy in this process.

Efficacy of a novel cervical dislocation tool for humane euthanasia of broilers and broiler breeders.

Euthanasia is an essential task performed daily on commercial poultry farms around the world to safeguard animal welfare. Manual cervical dislocation (MCD) is the most common euthanasia method but can be challenging to perform given the physical strength required to implement this technique. Therefore, the objective of this study was to evaluate the efficacy of a novel cervical dislocation tool (NCDT) compared to MCD. A total of 60 Ross 308 chickens (6-wk old) and 60 Ross 706 parent stock breeders (21-wk old) were enrolled in the study. Birds were sexed, blocked by body weight, and allocated to 1 of 2 treatments: 1) MCD and 2) NCDT. Immediately following euthanasia application, insensibility, and death were monitored. Once death was confirmed, gross evaluation, radiograph, and macroscopic/microscopic scoring were performed. Both euthanasia methods were 100% effective in achieving insensibility followed by cardiac and respiratory arrest in both age groups. In 6-wk-old broilers, there were no differences in insensibility measures or location and severity of the dislocation site by treatment. The NCDT treatment group showed an increased frequency of fractures located at the tooth-like process that projects from the cranial aspect of the centrum of the axis (dens) but had no impact on bird insensibility. For parent stock, differences in nictitating membrane reflex (NMR) and laceration scores for birds euthanized with NCDT were found and likely associated with additional force exerted with the tool. The NCDT is a promising replacement for MCD and future work should address the development of free and accessible training materials for on-farm use.

Clinical and imaging findings in dogs with nerve root signature associated with cervical intervertebral disc herniation.

BACKGROUND: Intervertebral disc herniation (IVDH) is the most common spinal cord disease in dogs. Little information is available regarding the clinical presentation of nerve root signature (NRS) associated with cervical IVDH. HYPOTHESIS/OBJECTIVE: To detail the clinical and magnetic resonance imaging (MRI) findings in dogs with NRS associated with cervical IVDH. ANIMALS: Forty-seven client-owned dogs presenting with thoracic limb NRS and MRI confirmed IVDH. METHODS: Medical records from 2010 to 2020 were retrospectively reviewed for dogs that met inclusion criteria. Imaging studies were evaluated by 2 individuals to characterize location and severity of neural tissue compression. RESULTS: Chondrodystrophoid dogs comprised the majority of the study cohort, with dachshund the most common breed (n = 10). Three-quarters of dogs were ≥7 years of age. Interobserver agreement was moderate or good for all of the imaging variables evaluated. The C6-C7 intervertebral disc space was significantly overrepresented (P = .01), comprising 32% (15/47) of the affected discs. However, 42% (20/47) of cases involved C2-C3 though C4-C5 disc sites. Disc material was more frequently located laterally compared to medially within the vertebral canal (P = .0005), and to be associated with compression of the nerve root at the level of the intervertebral foramen (P = .012). CONCLUSION/CLINICAL IMPORTANCE: NRS is most commonly associated with lateralized or foraminal cervical disc herniations. It is most prevalent with C6-C7 intervertebral disc involvement, suggesting that there might be unique anatomic factors that contribute to development of NRS at this site, but can be a clinical manifestation of IVDH occurring anywhere along the cervical spine.

Melon and rostral muscle morphology of Gervais' beaked whale (Mesoplodon europaeus): Alternating patterns of bilateral asymmetry.

Toothed whales utilize specialized nasal structures such as the lipid-rich melon to produce sound and propagate it into the aquatic environment. Very little nasal morphology of mesoplodont beaked whales has been described in the literature, and the anatomy of the melon and associated musculature of Gervais' beaked whale (Mesoplodon europaeus) remains undescribed. Heads of three (n = 3) Gervais' beaked whales were examined in detail via dissection as well as computed tomography (CT) and magnetic resonance imaging (MRI). Two additional Gervais' beaked whale individuals (n = 2) were studied via archived CT and MRI scans. Representative transverse dissection sections of the melon were processed for polarized light imaging to verify the presence of tendons inserting into the melon tissue. Three-dimensional (3D) CT reconstructions of the melon, rostral muscles, and associated structures were performed to assess morphology and spatial relationships. In all individuals, the melon's main body demonstrated a bilaterally asymmetrical, curvilinear geometry. This curvilinear shape was defined by a pattern of alternating asymmetry in the medial rostral muscles that projected into the melon's tissue. In transverse polarized light imaging, a network of tendons originating from these asymmetrical rostral muscle projections was observed permeating the melon's lipid tissue. This curvilinear melon morphology and associated asymmetrical musculature suggest a means of lengthening the lipid pathway within a relatively short dimensional footprint. In addition, the species-specific arrangement of muscular projections suggests complex fine-tuning of the melon's geometry during echolocation. Further studies may lend additional insight into the function of this unusual melon morphology.

Calculation of body surface area of corn snakes (Pantherophis guttatus) utilizing computed tomography.

OBJECTIVE: To determine the K-constant for body surface area calculation from body weight in corn snakes (Pantherophis guttatus) through the use of computed tomography (CT) measurements. ANIMALS: 12 adult corn snakes held by North Carolina State University for research purposes underwent CT between November 2022 and January 2023. METHODS: Each snake had a CT scan, physical examination, and body weight measurement. CT images were uploaded into software able to perform 3-D reconstruction and measure body surface area. The species-specific K-constant was determined using nonlinear regression analysis between body surface area and (body weight in grams)2/3. RESULTS: The mean body weight of the 12 adult corn snakes was 228 g, with a mean body surface area of 505.1 cm2. The calculated K-constant was 13.6 (P < .001). The resulting formula for body surface area in corn snakes is BSA in cm2 = 13.6 X (body weight in grams)2/3. CLINICAL RELEVANCE: The body surface area formula developed for corn snakes will allow for improved dosing accuracy for medications with low therapeutic safety margins. Additional pharmacokinetic and pharmacodynamic studies are necessary to determine the safety and efficacy of individual medications.

A comparison of the motor effects and analgesic efficacy following lumbar plexus block combined with sciatic nerve block or epidural in dogs undergoing tibial plateau leveling osteotomy.

OBJECTIVE: To compare motor effects and analgesic efficacy following an ultrasound-guided lateral approach to lumbar plexus blockade at L7 and sciatic nerve blockade (LPSNB) against epidural injection in dogs undergoing tibial plateau leveling osteotomy (TPLO). STUDY DESIGN: Prospective, randomized, blinded clinical trial. ANIMALS: A total of 27 healthy adult dogs undergoing unilateral TPLO surgery. METHODS: Dogs were allocated to either LPSNB (bupivacaine 2 mg kg-1, 0.75%) or epidural (morphine PF 0.1 mg kg-1 and bupivacaine 0.5 mg kg-1, 0.75%). Other aspects of clinical management were identical, including anesthetic drug protocol, area of presurgical clipping and bladder care. Time to perform the block, response to surgical stimuli, pain scores, rescue analgesia, time to stand and walk, motor score and time to first urination were recorded. One evaluator, unaware of treatment status, performed all evaluations. Student's t-test or Mann-Whitney U test was used to compare continuous variables between groups, and Fisher's exact test for categorical variables. RESULTS: Median (range) times to stand and walk were shorter for LPSNB [60 (40-120) minutes and 90 (60-150) minutes, respectively, p = 0.003] than for epidural [150 (120-240) minutes and 180 (120-360) minutes, respectively, p = 0.006]. Four dogs required rescue intraoperatively (three in epidural group, one in LPSNB group, p = 0.438). Pain scores over the 24 hour evaluation period were similar, and not significantly different, for each group. Time to spontaneous urination [LPSNB, 330 (240-360) minutes; epidural, 300 (120-1440) minutes, p = 1.0] did not differ between groups. CONCLUSIONS AND CLINICAL RELEVANCE: An ultrasound-guided lateral paravertebral approach to the lumbar plexus within the psoas compartment at L7, combined with sciatic nerve blockade, allows faster return to normal motor function, with similar pain control and impact on urination when compared with epidural in dogs after TPLO surgery.

Lumbar vertebral bone density is decreased in horses with pituitary pars intermedia dysfunction.

BACKGROUND: Pathological fractures have been reported in equids with pituitary pars intermedia dysfunction (PPID) but their prevalence and pathogenesis is unknown. OBJECTIVES: To compare: (1) bone mineral density (BMD) in weight bearing and nonweight bearing bones in PPID+ equids and aged and young PPID- controls; and (2) biomechanical properties of the fourth lumbar vertebral body in PPID+ equids and aged PPID- equids. STUDY DESIGN: Case-control study: five PPID+ equids and six aged and four young PPID- control horses. METHODS: PPID status was based on clinical signs and necropsy examination of the pituitary gland (PG). The lumbar vertebral column, right front third metacarpus (MC3), left hind third metatarsus (MT3), and PG were removed after euthanasia. BMD was determined by quantitative computed tomography of regions of interest (ROI) in each bone and biomechanical testing was performed on the fourth lumbar vertebral body. Serum concentrations of parathormone (PTH), ionised Ca++ , 25-hydroxyvitamin D, and osteocalcin (OC) were also measured. Data were analysed using one-way ANOVA and correlation analyses. RESULTS: BMD of trabecular and cortical regions of interest (ROI) of the third, fourth (L4), and fifth lumbar vertebrae were significantly lower in PPID+ equids as compared with aged (p < 0. 001) and young (p < 0.01) PPID- controls. In contrast, no differences were found in BMD of trabecular or cortical ROIs of MC3 and MT3 between groups. No differences were detected in force at fracture, displacement at fracture, Young's modulus or strain of L4 between PPID+ and aged PPID- horses. No differences were found in serum PTH, ionised Ca++ , 25-hydroxyvitamin D, or OC concentrations between groups. MAIN LIMITATIONS: Limited number of equids studied and variation in test results. CONCLUSIONS: BMD of nonweight bearing bones can be decreased with PPID and could increase risk of developing pathological fractures.

A short history of the phosphorus index and Andrew Sharpley's contributions from inception through development and implementation.

In the 1980s, growing recognition of agricultural phosphorus (P) sources to surface water eutrophication led to scrutiny of animal feeding operations. In 1990, the USDA-Natural Resources Conservation Service (NRCS) invited prominent scientists to find a solution. It was at an initial meeting that Dr. Andrew Sharpley suggested that P assessment could be modeled after the Universal Soil Loss Equation, where a matrix of factors influencing P loss would be associated with farm nutrient management recommendations. After codifying the P assessment into the USDA-NRCS 590 Nutrient Management Standard some 10 years later, 48 states chose to develop their own P Index. Sharpley, working with many others, helped develop several state P Indices. In 2000, Sharpley secured funding from the USDA-Agricultural Research Service to support the National P Research Project, which conducted in-field P runoff assessments using standardized rainfall simulated studies across 20 states; this allowed individual trials to be aggregated for agroecological regions that were then incorporated into specific state P Indices. Eventually, comparison of P Indices across state boundaries led to a white paper at the behest of USDA-NRCS that resulted in three regional projects evaluating modeling approaches to support or replace P Indices. Sharpley's national umbrella project pointed to shortcomings in water quality models, such as APEX or TBET, as a replacement for state P Indices, which remain a key part of the USDA-590 standard. As a selfless leader, capable of attracting and assembling diverse, productive interdisciplinary teams, Sharpley was essential to the inception, development, and implementation of the P Index.

Coupling and Slips in Photosynthetic Reactions-From Femtoseconds to Eons.

Photosynthesis stands as a unique biological phenomenon that can be comprehensively explored across a wide spectrum, from femtoseconds to eons. Across each timespan, a delicate interplay exists between coupling and inherent deviations that are essential for sustaining the overall efficiency of the system. Both quantum mechanics and thermodynamics act as guiding principles for the diverse processes occurring from femtoseconds to eons. Processes such as excitation energy transfer and the accumulation of oxygen in the atmosphere, along with the proliferation of organic matter on the Earth's surface, are all governed by the coupling-slip principle. This article will delve into select time points along this expansive scale. It will highlight the interconnections between photosynthesis, the global population, disorder, and the issue of global warming.

Defining a species-specific K constant in American Bullfrogs (Lithobates catesbeianus) through use of computed tomography.

OBJECTIVE: To use CT measurements to define the body surface area (BSA) formula in American bullfrogs (Lithobates catesbeianus) and calculate the species-specific shape constant (K) to suggest chemotherapeutic doses. ANIMALS: 12 American bullfrogs owned by the North Carolina State College of Veterinary Medicine Department of Laboratory Animal Resources underwent CT scans without anesthesia or sedation in November 2022. METHODS: As part of this prospective study, each American bullfrog underwent a complete physical exam and CT scan. 3-D surface models were created using CT data, and the resulting measurements were used for BSA calculations. Animals were grouped by sex. Nonlinear regression analysis of BSA versus body weight was performed, and a species-specific formula was derived for calculating BSA in American bullfrogs. RESULTS: The mean body weight of the bullfrogs was 354 grams. The mean CT-derived BSA was 414.92 cm2. The calculated K constant was 8.28 for the 12 American bullfrogs, and the CT-derived BSA formula was BSA in cm2 = 8.28 X (body weight in g)2/3. The K constant was 8.07 for females and 8.44 for males and was not significantly different between sexes (P = .5). CLINICAL RELEVANCE: Results indicated that the species-specific K constant for American bullfrogs is 8.28. This is the first calculated K constant that exists for amphibians to our knowledge.

Structure of Chlorella ohadii Photosystem II Reveals Protective Mechanisms against Environmental Stress.

Green alga Chlorella ohadii is known for its ability to carry out photosynthesis under harsh conditions. Using cryogenic electron microscopy (cryoEM), we obtained a high-resolution structure of PSII at 2.72 Å. This structure revealed 64 subunits, which encompassed 386 chlorophylls, 86 carotenoids, four plastoquinones, and several structural lipids. At the luminal side of PSII, a unique subunit arrangement was observed to protect the oxygen-evolving complex. This arrangement involved PsbO (OEE1), PsbP (OEE2), PsbB, and PsbU (a homolog of plant OEE3). PsbU interacted with PsbO, PsbC, and PsbP, thereby stabilizing the shield of the oxygen-evolving complex. Significant changes were also observed at the stromal electron acceptor side. PsbY, identified as a transmembrane helix, was situated alongside PsbF and PsbE, which enclosed cytochrome b559. Supported by the adjacent C-terminal helix of Psb10, these four transmembrane helices formed a bundle that shielded cytochrome b559 from the surrounding solvent. Moreover, the bulk of Psb10 formed a protective cap, which safeguarded the quinone site and likely contributed to the stacking of PSII complexes. Based on our findings, we propose a protective mechanism that prevents QB (plastoquinone B) from becoming fully reduced. This mechanism offers insights into the regulation of electron transfer within PSII.

CryoEM PSII structure reveals adaptation mechanisms to environmental stress in Chlorella ohadii.

Performing photosynthesis in the desert is a challenging task since it requires a fast adaptation to extreme illumination and temperature changes. To understand adaptive mechanisms, we purified Photosystem II (PSII) from Chlorella ohadii , a green alga from the desert soil surface, and identified structural elements that might enable the photosystem functioning under harsh conditions. The 2.72 Å cryogenic electron-microscopy (cryoEM) structure of PSII exhibited 64 subunits, encompassing 386 chlorophylls, 86 carotenoids, four plastoquinones, and several structural lipids. At the luminal side of PSII, the oxygen evolving complex was protected by a unique subunit arrangement - PsbO (OEE1), PsbP (OEE2), CP47, and PsbU (plant OEE3 homolog). PsbU interacted with PsbO, CP43, and PsbP, thus stabilising the oxygen evolving shield. Substantial changes were observed on the stromal electron acceptor side - PsbY was identified as a transmembrane helix situated alongside PsbF and PsbE enclosing cytochrome b559, supported by the adjacent C-terminal helix of Psb10. These four transmembrane helices bundled jointly, shielding cytochrome b559 from the solvent. The bulk of Psb10 formed a cap protecting the quinone site and probably contributed to the PSII stacking. So far, the C. ohadii PSII structure is the most complete description of the complex, suggesting numerous future experiments. A protective mechanism that prevented Q B from rendering itself fully reduced is proposed.

Uncertainty in phosphorus fluxes and budgets across the US long-term agroecosystem research network.

Phosphorus (P) budgets can be useful tools for understanding nutrient cycling and quantifying the effectiveness of nutrient management planning and policies; however, uncertainties in agricultural nutrient budgets are not often quantitatively assessed. The objective of this study was to evaluate uncertainty in P fluxes (fertilizer/manure application, atmospheric deposition, irrigation, crop removal, surface runoff, and leachate) and the propagation of these uncertainties to annual P budgets. Data from 56 cropping systems in the P-FLUX database, which spans diverse rotations and landscapes across the United States and Canada, were evaluated. Results showed that across cropping systems, average annual P budget was 22.4 kg P ha-1 (range = -32.7 to 340.6 kg P ha-1 ), with an average uncertainty of 13.1 kg P ha-1 (range = 1.0-87.1 kg P ha-1 ). Fertilizer/manure application and crop removal were the largest P fluxes across cropping systems and, as a result, accounted for the largest fraction of uncertainty in annual budgets (61% and 37%, respectively). Remaining fluxes individually accounted for <2% of the budget uncertainty. Uncertainties were large enough that determining whether P was increasing, decreasing, or not changing was inconclusive in 39% of the budgets evaluated. Findings indicate that more careful and/or direct measurements of inputs, outputs, and stocks are needed. Recommendations for minimizing uncertainty in P budgets based on the results of the study were developed. Quantifying, communicating, and constraining uncertainty in budgets among production systems and multiple geographies is critical for engaging stakeholders, developing local and national strategies for P reduction, and informing policy.

Structure of Photosystem I Supercomplex Isolated from a Chlamydomonas reinhardtii Cytochrome b6f Temperature-Sensitive Mutant.

The unicellular green alga, Chlamydomonas reinhardtii, has been widely used as a model system to study photosynthesis. Its possibility to generate and analyze specific mutants has made it an excellent tool for mechanistic and biogenesis studies. Using negative selection of ultraviolet (UV) irradiation-mutated cells, we isolated a mutant (TSP9) with a single amino acid mutation in the Rieske protein of the cytochrome b6f complex. The W143R mutation in the petC gene resulted in total loss of cytochrome b6f complex function at the non-permissive temperature of 37 °C and recovery at the permissive temperature of 25 °C. We then isolated photosystem I (PSI) and photosystem II (PSII) supercomplexes from cells grown at the non-permissive temperature and determined the PSI structure with high-resolution cryogenic electron microscopy. There were several structural alterations compared with the structures obtained from wild-type cells. Our structural data suggest that the mutant responded by excluding the Lhca2, Lhca9, PsaL, and PsaH subunits. This structural alteration prevents state two transition, where LHCII migrates from PSII to bind to the PSI complex. We propose this as a possible response mechanism triggered by the TSP9 phenotype at the non-permissive temperature.

General anesthesia does not induce ultrasonographic changes in the pleura of healthy adult horses anesthetized for elective magnetic resonance imaging.

OBJECTIVE: To compare thoracic ultrasonographic findings in healthy horses before and after general anesthesia for elective MRI utilizing a recently developed ultrasonographic scoring system to aid clinicians in the early identification of pneumonia following anesthesia. ANIMALS: 13 adult horses > 3 years of age. PROCEDURES: Prior to anesthesia, horses underwent a thorough physical examination, CBC, thoracic radiography, and thoracic ultrasonography. Horses were then anesthetized for elective MRI, and thoracic ultrasonography was repeated within 3 hours after recovery. Thoracic ultrasonographic findings were scored utilizing a recently developed scoring system, and scores were compared before and after anesthesia. RESULTS: There was no significant difference identified in total thoracic ultrasonography score before and after anesthesia, and there was no correlation between thoracic ultrasonography score following anesthesia and the body weight of the horse, the time recumbent, and the dependent side. CLINICAL RELEVANCE: In healthy horses undergoing anesthesia for elective imaging, there was no significant change in thoracic ultrasonographic findings 3 hours after recovery from anesthesia. These data can aid clinicians in determining the clinical significance of ultrasonographic changes in the lung in the immediate postanesthetic period.

Structure of Dunaliella photosystem II reveals conformational flexibility of stacked and unstacked supercomplexes.

Photosystem II (PSII) generates an oxidant whose redox potential is high enough to enable water oxidation , a substrate so abundant that it assures a practically unlimited electron source for life on earth . Our knowledge on the mechanism of water photooxidation was greatly advanced by high-resolution structures of prokaryotic PSII . Here, we show high-resolution cryogenic electron microscopy (cryo-EM) structures of eukaryotic PSII from the green alga Dunaliella salina at two distinct conformations. The conformers are also present in stacked PSII, exhibiting flexibility that may be relevant to the grana formation in chloroplasts of the green lineage. CP29, one of PSII associated light-harvesting antennae, plays a major role in distinguishing the two conformations of the supercomplex. We also show that the stacked PSII dimer, a form suggested to support the organisation of thylakoid membranes , can appear in many different orientations providing a flexible stacking mechanism for the arrangement of grana stacks in thylakoids. Our findings provide a structural basis for the heterogenous nature of the eukaryotic PSII on multiple levels.

Agronomic and economic implications of cover crop and phosphorus fertilizer management practices for water quality improvement.

Phosphorus (P) fertilization practices and winter cover crops are promoted to protect water quality yet can potentially influence crop yield and profitability. This study examined the impacts of three P fertilizer management practices (no P, fall broadcast P, and spring injected P) and winter annual cover crop use on yields, net returns, and water quality in a no-till corn-soybean rotation. Treatments were replicated in a 4-yr field study where sediment, total P, and dissolved reactive P (DRP) losses in edge-of-field surface runoff were continuously monitored. Production budgets were developed for each treatment and coupled with water quality data to identify optimal practices and opportunity costs for improving water quality. Applying P fertilizers increased crop yield regardless of application method or cover crop, but the response was more pronounced in corn than soybean. The cover crop reduced corn grain yield in 1 yr with high temperature and low precipitation but did not impact grain yield in the other corn year or either year of soybean. The most profitable treatment was fall broadcast P fertilizer with no cover crop, which also had the greatest total P and DRP losses and near greatest sediment loss. The lowest-cost methods of reducing total P, DRP, and sediment losses were, respectively, no P fertilizer without a cover crop ($47.56 kg-1 total P), spring injected P fertilizer without a cover crop ($56.47 kg-1 DRP), and spring injected P fertilizer with a cover crop ($0.16 kg-1 sediment). Widespread adoption of these practices will likely require monetary incentives.