Cloning and pharmacological characterization of the equine adenosine A2A receptor: a potential therapeutic target for the treatment of equine endotoxemia

The aim of the current study was to clone the equine adenosine A(2A) receptor gene and to establish a heterologous expression system to ascertain its pharmacologic profile via radioligand binding and functional assays. An eA(2A)-R expression construct was generated by ligation of the eA(2A) cDNA into the pcDNA3.1 expression vector, and stably transfected into human embryonic kidney cells (HEK). Binding assays identified those clones expressing the eA(2A)-R, and equilibrium saturation isotherm experiments were utilized to determine dissociation constants (K(D)), and receptor densities (B(max)) of selected clones. Equilibrium competition binding revealed a rank order of agonist potency of ATL > CV-1808 > NECA > 2-CADO > CGS21680, and a rank order of antagonist potency as ZM241385 > 8-phenyltheophylline > p-sulfophenyltheophylline > caffeine. Furthermore, adenylate cyclase assays using selective A(2A)-R agonists revealed that the eA(2A)-R functionally coupled to Galpha(s) as indicated by an increase in intracellular [(3)H]cAMP upon receptor activation. Finally, NF-kappaB reporter gene assays revealed a CGS21680 concentration-dependent inhibition of NF-kappaB activity. These results indicate that the heterologously expressed eA(2A)-R has a pharmacological profile similar to that of other mammalian A(2A) receptors and thus can be utilized for further characterization of the eA(2A)-R to ascertain whether it can serve as a suitable pharmacological target for equine inflammatory disease.     

Screening of the entire USDA castor germplasm collection for oil content and fatty acid composition for optimum biodiesel production

Castor has tremendous potential as a feedstock for biodiesel production. The oil content and fatty acid composition in castor seed are important factors determining the price for production and affecting the key fuel properties of biodiesel. There are 1033 available castor accessions collected or donated from 48 countries worldwide in the USDA germplasm collection. The entire castor collection was screened for oil content and fatty acid composition by nuclear magnetic resonance (NMR) and gas chromatography (GC), respectively. Castor seeds on the average contain 48.2% oil with significant variability ranging from 37.2 to 60.6%. Methyl esters were prepared from castor seed by alkaline transmethylation. GC analysis of methyl esters confirmed that castor oil was composed primarily of eight fatty acids: 1.48% palmitic (C16:0), 1.58% stearic (C18:0), 4.41% oleic (C18:1), 6.42% linoleic (C18:2), 0.68% linolenic (C18:3), 0.45% gadoleic (C20:1), 84.51% ricinoleic (C18:1-1OH), and 0.47% dihydroxystearic (C18:0-2OH) acids. Significant variability in fatty acid composition was detected among castor accessions. Ricinoleic acid (RA) was positively correlated with dihydroxystearic acid (DHSA) but highly negatively correlated with the five other fatty acids except linolenic acid. The results for oil content and fatty acid composition obtained from this study will be useful for end-users to explore castor germplasm for biodiesel production.     

An Assessment of State-and-Transition Models: Perceptions Following Two Decades of Development and Implementation

State-and-transition models (STMs) are being developed for many areas in the United States and represent an important tool for assessing and managing public and private rangelands. Substantial resources have been invested in model development, yet minimal efforts have been made to evaluate the utility of STMs for rangeland assessment and management. We interviewed 47 rangeland professionals, equally divided between managers and researchers, in four ecoregions to determine their perceptions of the purpose, development, and strengths and weaknesses of STMs to assess the status of the STM framework. Our analysis identified three primary perspectives regarding the purpose of STMs: a decision-making tool for land managers, a means to represent the complex dynamics of rangeland ecosystems, and an effective communication tool. These diverse views of STM purposes were associated with differing perspectives concerning model development that identified five major issues in need of further development and refinement: 1) the relative importance of management practices and ecological processes in driving transitions, 2) the criteria used to define thresholds, 3) the appropriate level of model complexity, 4) the respective roles of expert knowledge and ecological data in model development, and 5) processes for model review and revision. We recommend greater dialogue among researchers and managers to further clarify STM terminology and develop standard protocols for model development and validation. Mechanisms are critically needed to assure peer review and revision of existing models so that STMs are continually updated to reflect current understanding of rangeland dynamics.     

Potential influence of sugarcane cultivation on estuarine water quality of Louisiana's gulf coast

Sugarcane is cultivated on some 170000 ha of land in south central and southwestern Louisiana. This acreage drains into bayous and rivers that empty into Louisiana's coastal bays and estuaries. For a number of years the state's Department of Agriculture and Forestry and Department of Environmental Quality have collected water quality data from this sugarcane area. Study of these data shows that approximately one in five detections of atrazine is above the maximum contaminant level (MCL) for drinking water. Currently there is no U.S. atrazine standard for protection of aquatic life. February and October detections of this herbicide are probably due to sugarcane cultivation. Nitrate levels have remained below the MCL for drinking water, but nitrate and phosphorus concentrations may pose a potential for eutrophication problems. The contribution of sugarcane production to the nutrient status of Louisiana's coastal water bodies is difficult to assess because there are other sources of nutrients in the area and native soil phosphorus levels are high. Cultural practices such as subsurface drains, open drainage ditches, and postharvest residue management have potential through enhancement of soil infiltration for decreasing sugarcane's contribution to water quality problems in southern and coastal Louisiana. A new field project is being installed at the Louisiana State University Agricultural Experiment Station's Sugarcane Research Station at St. Gabriel to assess the water quality benefits of these practices with respect to sugarcane cultivation.     

An updated geographic distribution of Myxobolus cerebralis (Hofer, 1903) (Bivalvulida: Myxobolidae) and the first diagnosed case of whirling disease in wild-caught trout in the south-eastern United States

Myxobolus cerebralis (Bivalvulida: Myxobolidae), the aetiological agent of salmonid whirling disease, was detected in 2 river basins of North Carolina during 2015, which initiated the largest spatial–temporal monitoring project for the disease ever conducted within the south-eastern United States (focused mainly in eastern Tennessee and western North Carolina). A total of 2072 rainbow trout Oncorhynchus mykiss, 1,004 brown trout Salmo trutta and 468 brook trout Salvelinus fontinalis were screened from 113 localities within 7 river basins during June 2017 through October 2019. Infections were detected by pepsin–trypsin digest, microscopy and the species-specific nested polymerase chain reaction (PCR) in 19 localities across 6 river basins. Myxospore morphology was indistinguishable from the published literature. In 2019, five rainbow trout that symptomatic for whirling disease (sloping neurocranium and lordosis) were captured and processed for histopathology. Myxospores were detected in the calvarial cartilage of two deformed trout with associated erosion of the cartilage consistent with reported whirling disease lesions. This is the first report of M. cerebralis in Tennessee and the first histologically confirmed cases of whirling disease in southern Appalachian (south-eastern United States) rivers and streams and expands the distribution of M. cerebralis throughout western North Carolina and eastern Tennessee.     

Opportunities for agent‐based modelling in human dimensions of fisheries

Models of human dimensions of fisheries are important to understanding and predicting how fishing industries respond to changes in marine ecosystems and management institutions. Advances in computation have made it possible to construct agent‐based models (ABMs)—which explicitly describe the behaviour of individual people, firms or vessels in order to understand and predict their aggregate behaviours. ABMs are widely used for both academic and applied purposes in many settings including finance, urban planning and the military, but are not yet mainstream in fisheries science and management, despite a growing literature. ABMs are well suited to understanding emergent consequences of fisher interactions, heterogeneity and bounded rationality, especially in complex ecological, social and institutional contexts. For these reasons, we argue that ABMs of human behaviour can contribute significantly to human dimensions of fisheries in three areas: (a) understanding interactions between multiple management institutions; (b) incorporating cognitive and behavioural sciences into fisheries science and practice; and (c) understanding and projecting the social consequences of management institutions. We provide simple examples illustrating the potential for ABMs in each of these areas, using conceptual (“toy”) versions of the POSEIDON model. We argue that salient strategic advances in these areas could pave the way for increased tactical use of ABMs in fishery management settings. We review common ABM development and application challenges, with the aim of providing guidance to beginning ABM developers and users studying human dimensions of fisheries.     

Presence of the Streptococcus suis suilysin gene and expression of MRP and EF correlates with high virulence in Streptococcus suis type 2 isolates

Nineteen Streptococcus suis type 2 isolates that had been analyzed previously for hemolysin production, ribotype, and virulence in pigs were examined for presence of the gene coding for suilysin by PCR amplification, and southern blot and hybridization techniques. Based on southern blot and hybridization analysis, all isolates tested contained at least a portion of the suilysin gene. PCR amplification of the entire gene resulted in gene fragments from five of the seven highly virulent isolates and none of the moderately virulent or avirulent isolates. Additional PCR analysis showed that mutation or deletions at the 5′ end of the suilysin gene in the less virulent isolates prevented amplification of the sly gene fragment from those isolates. The MRP+ (muramidase-released protein) EF+ (extracellular protein) phenotype was also expressed by the same five highly virulent/sly+ isolates.