Nitrate uptake by Enteromorpha spp. (Chlorophyceae): Applications to aquaculture systems

Enteromorpha linza and E. intestinalis thrive in aquatic systems rich in inorganic fixed nitrogen. At 15°C and ca. 1800 μW cm^?2 these algae showed a V_max for nitrate uptake of 129 μmol h^? g^?1 dry weight ( = ca. 22 μmol h^?1 g^?1 wet weight) and a K_s of 17 μM. At 20 μM rates averaged 70 ± 21 μmol h^?1 g^?1. They dropped to 19% of this value at 5°C, and at 0°C nitrate uptake ceased. In the first hour of darkness, nitrate uptake decreased to 35% and in the second hour to 10% of the rate in light. Enteromorpha spp. scavenged nitrate from the medium at concentrations < 0.5 μM, and no nitrate leaked out of the tissue into nitrate-free water. Information from this study may be useful in designing nutrient feeding strategies in aquaculture systems.     

Preparation and Characterization of Starch Particles for Use in Pickering Emulsions

Particle‐stabilized emulsions, called Pickering emulsions, can be produced by using starch particles. In this work we studied how the properties of the starch particles affect the droplet size and creaming of such emulsions. In the study, various sizes of starch particles were generated by two different methods and used to stabilize Pickering emulsions. Sedimentation according to Stokes’ law was used to separate small and large starch granules. Acid hydrolysis was another method used to obtain smaller particles. All samples were modified with octenyl succinic anhydride (OSA) to increase their hydrophobicity with a level of OSA substitution between 1.8 and 3.1%. The size of starch particles was the main factor influencing emulsion droplet sizes. Furthermore, the droplet size decreased as the starch concentration increased. Using small starch particles with sizes <10 μm produced stable emulsions with smaller droplet size compared with larger sizes of starch particles, >10 μm. When subjected to acid hydrolysis, smaller starch particles were generally obtained, which could subsequently create smaller emulsion droplets. The emulsion index increased for the acid‐hydrolyzed starch owing to the size reduction of starch particles. The shape of the starch seemed to have a minor impact on the droplet size and the creaming of Pickering emulsions.     

Protein electrophoresis of psittacine plasma

BACKGROUND: Although protein electrophoresis (EPH) has been widely applied in human and veterinary medicine, it has only recently been implemented in the analysis of avian samples. OBJECTIVE: The purpose of this study was to examine the application of protein EPH to the analysis of psittacine plasma samples. Our goals were to describe protein fraction mobility, establish reference intervals for some common species, determine the coefficient of variation (CV) of the chosen method, and examine the effects of sample handling and sample condition. METHODS: Heparinized plasma samples from several common psittacine species (minimum sample size 50 each) were examined using the Beckman Paragon system and SPEP-II gels. Total protein was measured by refractometry. Reference intervals (95%) were calculated by the rank methods. RESULTS: Fraction migration patterns were found to vary among common psittacine species. Day-to-day CV for the EPH fractions ranged from 2.2% to 10.5%; within-run CV ranged from 4.8% to 10.8%; and total CV ranged from 3.2% to 14.8%. The highest CV was noted for the poorly defined alpha-globulin fraction. Prolonged refrigeration, repeated freeze-thawing, hemolysis, and lipemia altered the results. CONCLUSIONS: Protein fractions from psittacine species were variable in terms of migration pattern and protein concentration, which necessitates the use of species-specific reference intervals. Avian protein electrophoretic patterns and values should be interpreted based on knowledge of the CV associated with the technique as well as on the effects of sample handling and condition.     

Lobomycosis in Atlantic bottlenose dolphins from the Indian River Lagoon, Florida

OBJECTIVE: To determine the prevalence of lobomycosis, a mycotic infection of dolphins and humans caused by a yeastlike organism (Lacazia loboi), among dolphins in the Indian River Lagoon in Florida. DESIGN: Cross-sectional study. ANIMALS: 146 Atlantic bottlenose dolphins. PROCEDURE: Comprehensive health assessments of bottlenose dolphins in the Indian River Lagoon of Florida (n = 75) and in estuarine waters near Charleston, SC (71), were conducted during 2003 and 2004. Bottlenose dolphins were captured, examined, and released. Skin lesions were photographed and then biopsied. Tissue sections were stained with H&E and Gomori methenamine silver stains for identification of L. loboi. RESULTS: 9 of 30 (30%) dolphins captured in the southern portion of the Indian River Lagoon had lobomycosis, whereas none of the 45 dolphins captured in the northern portion of the lagoon or of the 71 dolphins captured near Charleston, SC, did. Affected dolphins had low serum alkaline phosphatase activities and high acute-phase protein concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that lobomycosis may be occurring in epidemic proportions among dolphins in the Indian River Lagoon. Localization of the disease to the southern portion of the lagoon, an area characterized by freshwater intrusion and lower salinity, suggests that exposure to environmental stressors may be contributing to the high prevalence of the disease, but specific factors are unknown. Because only dolphins and humans are naturally susceptible to infection, dolphins may represent a sentinel species for an emerging infectious disease.     

Isolation and characterization of factor I of the bovine complement system

OBJECTIVE: To isolate and characterize factor I of the bovine complement system. Sample Population-Serum obtained from the blood of beef cattle. PROCEDURES: Serum samples were fractionated to yield factor I by means of sequential precipitation, ion-exchange, and gel-filtration chromatography. The protein was identified throughout the procedure on the basis of its ability to degrade the alpha'-chain of bovine C3b in the presence of bovine factor H. Electrophoresis in polyacrylamide gels was used to assess the degradation of C3b and determine the molecular weights of factor I and its component polypeptide chains. RESULTS: Bovine factor I had an apparent molecular weight of 94 kd and consisted of 2 disulfide-bonded polypeptides that had apparent molecular weights of 51 and 42 kd (under reducing conditions). Factor H was required for the factor I cleavage of the alpha'-chain of bovine C3b into iC3b. A similar cofactor effect was provided by trypsinized bovine erythrocytes or erythrocyte ghosts. Bovine properdin was prepared and shown to be a single polypeptide chain of 58 kd in the reduced form. CONCLUSIONS AND CLINICAL RELEVANCE: Bovine factor I can be purified from serum by a simple 4-step procedure. It is structurally and functionally comparable to factor I of other species, and its purification completes the isolation and characterization of all the soluble components of the bovine alternative complement pathway.     

Isolation and characterization of the third component of bovine complement

C3 was obtained from bovine serum by polyethylene glycol precipitation and chromatography on DEAE-Sephadex A-50, CM-Sephadex A-50 and Sephacryl S-200. The protein has a molecular weight of 183,000 (alpha-chain 114,000 and beta-chain 69,000). A CVF-induced bovine C3 convertase (Sepharose-CVF.Bb) cleaved C3 into C3a (11,000) and C3b (172,000) as shown by SDS-polyacrylamide gel electrophoresis. Isoelectricfocusing of C3 demonstrated at least three electrophoretic variants with pI 6.55–6.85. The isolated protein promoted the formation and action of a C3 convertase in the presence of purified bovine factors B and D. A monospecific antiserum prepared in rabbits failed to cross react with human C3 or CVF. C3c was identified as a contaminant during the isolation of C3.     

Monitoring unfractionated heparin therapy in dogs by measuring thrombin generation

Background: The calibrated automated thrombogram (CAT), an assay that permits measurement of thrombin generation in plasma, may be useful in studying hemostatic disorders and anticoagulant therapy in animals. Objectives: The aims of the study were to measure thrombin generation in healthy Beagle dogs and to evaluate the potential use of the CAT assay for monitoring therapy with unfractionated heparin (UFH). Methods: Individual platelet‐poor plasma samples and a plasma pool from 20 healthy adult Beagles were prepared. Serial UFH plasma dilutions were used to establish an in vitro heparin‐sensitivity curve. The pharmacodynamic effects of heparin in vivo were evaluated in Beagles using the CAT assay to measure thrombin generation with tissue factor at a concentration of 5 pM for initiation. Results: In healthy Beagles, the range of endogenous thrombin potential (ETP) was 238.7–414.0 nM/min (mean ± SD, 340.4 ± 63.1 nM/min). ETP intra‐assay and interassay variations were 7.1% and 12.9%, respectively. In vitro, a UFH concentration ≥0.4 U/mL resulted in total inhibition of thrombin generation. In vivo, the maximal effect of UFH on ETP was observed at 170 ± 36 minutes (range, 120–210 minutes) and resulted in a decrease in ETP of 38.5 ± 7.8% (range, 26.5–50.3%). In 210–420 minutes, ETP returned to baseline in 5 dogs. Conclusion: Our study demonstrates that thrombin generation can be measured in canine plasma and may be useful in assessing the degree of anticoagulation provided by UFH.     

Effects of theophylline on tracheal mucociliary clearance rates in healthy cats

OBJECTIVE: To determine tracheal mucociliary clearance rate (TMCCR) by use of a standard protocol in healthy anesthetized cats and to determine the effect of theophylline on TMCCR in healthy anesthetized cats. ANIMALS: 6 healthy cats. PROCEDURE: Cats were anesthetized with propofol, and a droplet of the radiopharmaceutical technetium Tc 99m macroaggregated albumin was placed endoscopically at the carina. Dynamic acquisition scintigraphic imaging was performed, using the larynx as the end point. The TMCCR was determined by measuring the distance the droplet traveled by frame rate. Each cat was imaged 6 times as follows: 3 times following placebo administration and 3 times following the administration of sustained release theophylline (25 mg/kg, PO). Serum theophylline concentrations were assessed during imaging to ensure therapeutic concentrations. RESULTS: The TMCCR in healthy adult cats anesthetized with propofol was 22.2 +/- 2.8 mm/min. Tracheal mucociliary clearance rate in cats receiving theophylline was 21.8 +/- 3.5 mm/min. Theophylline administration did not significantly alterTMCCR. CONCLUSIONS AND CLINICAL RELEVANCE: Theophylline has been shown to increase TMCCR in humans and dogs. In our study, we determined TMCCR in healthy anesthetized cats and found that it was not accelerated by the administration of theophylline.