Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Metal toxicosis in horses.

The ubiquity and stability of metals in the environment make them unique as a pollutant or an essential dietary component. Metals are neither created nor destroyed by chemical processes but are redistributed in the environment. In combination with other elements, metal compounds and alloys are essential materials of the contemporary world. Inappropriate use or distribution in the environment leads to adverse health effects on all biologic systems, including horses. Gastrointestinal upset is a common feature of acute toxicosis with metals in general. Among the metals discussed, arsenic and inorganic mercury have a propensity to do severe damage to the gut. Deposition of cadmium on forage is the source most likely to intoxicate horses. This subchronic to chronic problem in horses is manifest as disease of the musculoskeletal system and kidneys. Iron-containing hematinics are widely used in racetrack horses and occasionally result in hepatopathy when excessive doses are administered. Lead continues to be recognized as the most significant environmental metal pollutant. Poisoning is encountered routinely in humans and animals. Of the animal species of veterinary concern, lead-poisoned horses are not a frequent encounter. Lead-intoxicated horses show signs of peripheral neuropathy (laryngeal hemiplegia), intermittent colic, and mild anemia. Acute mercury poisoning sometimes occurs from the common use of mercury-containing blistering agents, with most clinical findings related to acute renal failure. Chronic excessive intake of zinc by horses is uncommon but devastating in rapidly growing foals. The mechanism of chronic zinc toxicosis is coupled to the induced copper deficiency. The condition is a disease of cartilage in the articular and growth physes.     

Surgical treatment of lameness.

Digital diseases are commonly seen in cattle. Cattle affected by digital disorders do not always respond to conservative therapy and require surgery. Surgical procedures for the common digital disorders are described, with emphasis on the different approaches to the distal interphalangeal joint.     

Quantitation of adenine nucleotides in equine colonic mucosal tissue using high performance liquid chromatography.

The objectives were to use high performance liquid chromatography (HPLC) to validate an established method for adenine nucleotide separation in equine colonic mucosal tissue, to determine the inherent variability in the tissue and extraction method, and to determine the stability of ATP, ADP, and AMP in the tissue with time. Equine colonic mucosal tissue obtained from a single horse was immediately submersed in liquid nitrogen, and stored at -70 degrees C. Samples were lyophilized, extracted, and separated by HPLC. The limit of quantitation was 0.05 microg/mL. The coefficient of variation for the instrument was less than 10% for all nucleotides measured. When the tissue was not homogenized prior to sampling, there were significant differences in adenine nucleotide content between samples. However, when the tissue was homogenized prior to analysis, these differences were no longer significant. There was no significant decrease in ATP, ADP, or AMP content over a 54-day analysis period.     

The Effect of Diet on the Biochemical Composition of Juvenile Artemia: Potential Formulations for Rock Lobster Aquaculture

The lipid class and fatty acid (FA) composition of juvenile Artemia fed continuously on four diets—the microalga Tetraselmis suecica , a mix of oat bran-wheat germ-lecithin (OWL), OwL-eicosapentaenoic acid (EPA), and OWL-EPA-arachidonic acid (AA)—were examined over a 9-d experiment in an attempt to approximate the FA profile of phyllosoma larvae of wild southern rock lobster Jasus edwardrii . The main difference in lipid class composition of Artemia fed the four diets was the relative level of polar lipid (PL) and triacylglycerol (TAG). By day 9, the algal-fed Artemia were highest in PL (95% of total lipid) and lowest in TAG (2%), whereas the remaining diets resulted in Artemia with 16–30% PL and 41–82% TAG. After 2 d, the relative FA composition of all Artemia treatments closely reflected those of the diets, with no marked change after further feeding (to day 9). In terms of the content of essential polyunsaturated fatty acids (PUFA), by day 5 Artemia fed: 1) with the algal diet contained 7 mg/g FA dry mass (0.3% DHA, 6.3% EPA, 3.4% AA of total FA); 2) with the OWL diet contained 3 mg/g (0.3% DHA, 0.9% EPA, 0.7% AA); 3) with the OWL-EPA diet contained 55 mg/g (6.2% DHA, 11.6% EPA, 1.1% AA); and 4) with the OWL-EPA-AA contained 83 mg/g (3.8% DHA, 7.5% EPA, 17.4% AA). The PUFA profiles of Artemia using the OWL-oil diets were similar to wild rock lobster phyllmmata, although levels of doco-sahexaenoic acid (DHA) were lower (10% DHA) than in J. edwardsii larvae. On the basis of PUFA composition data alone, the results suggest the suitability of the OWL-oil mixed diets for consideration for feeding to Artemia used in the culture of southern rock lobster larvae, particularly if the level of DHA can be further enhanced.     

Radiographic and ultrasonographic features of uroliths and other urinary tract filling defects

Radiopaque uroliths and nonradiopaque (water density) uroliths are filling defects encountered in the urinary tracts of dogs and cats. Other free luminal and attached soft tissue density filling defects encountered during uroradiographic special procedures include blood clots, air bubbles, hematomas, granulomas, abscesses, inflammatory and neoplastic polyps. Nonradiopaque uroliths cannot be identified on survey radiographs from other soft tissue dense structures. Gray scale ultrasonography can be used to differentiate nonradiopaque (water dense) uroliths from other soft tissue attached or free luminal filling defects of the excretory pathway. The differential radiographic features of filling defects encountered during cystography and urethrography are described and illustrated.     

Dietary crude protein concentration does not affect the leucine requirement of growing dogs

The objective of the present study was to examine the interaction between graded levels of leucine and dietary crude protein. Dose–response curves were generated using four 3 × 3 Latin squares (two dogs/square). Each square represented one of two concentrations of crude protein (140 or 280 g/kg diet) and one of two combinations of three concentrations of leucine (5.0, 7.0 and 9.0 g/kg diet or 9.0, 11 and 13 g/kg diet). An additional experiment was performed by feeding crude protein at 210 g/kg diet with either 7.0 or 11 g leucine/kg diet. Weight gain, food intake, nitrogen retention, plasma albumin and plasma amino acids were measured. The requirement was determined to be the minimum leucine concentration required to maximize weight gain and nitrogen retention. For 8–14-week-old male Beagle dogs, 140 g crude protein/kg diet in a diet containing 18 kJ metabolizable energy/g does not appear to support maximal growth. The leucine requirement was not affected by doubling the dietary crude protein level from 140 to 280 g/kg diet. From these results, the leucine requirement of 8–14-week-old Beagle dogs appears to be 11 g leucine/kg diet independent of the level of dietary crude protein, whereas dogs over 14 weeks require only 7 g leucine/kg diet for maximal nitrogen retention.