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.     

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.     

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.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

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.     

Essential Fatty Acids and the Cat

Abstract— The physiology of essential fatty acid metabolism in the cat is reviewed. Emphasis is placed on those aspects of the n:6 and n:3 fatty acids, their metabolites and interactions, which relate primarily to the skin. The functional roles, if known, of the fatty acids are discussed. Recent clinical research into the use of essential fatty acid supplements in the management of feline dermatoses is presented. Current indications for the therapeutic supplementation with essential fatty acids are summarised.     

Corneocyte Counts for Evaluation of Antiseborrheic Shampoos in Dogs

Abstract— An objective method for measuring corneocyte numbers before and after antiseborrheic shampoo therapy was assessed in dogs. Corneocytes were collected from six skin regions on the dorsal trunk of nine colony-raised beagles with clinically normal skin. Forty-eight collection sites were randomly assigned a treatment with one of seven medicated shampoos or water alone; six additional sites served as non-treated controls. Corneocytes were collected immediately before the first treatment, and 14 and 28 days after initiation of therapy with a 0.1% Tween 80 surfactant and a quantitative cup-scrub technique. Pretreatment corneocyte counts were not significantly different among the nine dogs nor among the randomly assigned treatment and control sites (3.55 · 10⁴ cells.cm^-2, SEM 0.17). Corneocyte counts increased in all treatment and control sites from days 0 to 14. Corneocyte counts were similar in the non-treated and water-treated sites from days 14 to 28. Corneocyte counts were not significantly different when the same treatments were compared on days 0, 14 and 28, or when different treatments were compared on the same day. The corneocyte collection technique used in this study proved to be a reliable method for assessing the rate of cell desquamation and cell surface cohesion in dogs.     

Factors affecting the reproductive performance of the weaned sow.

The majority of sows return to estrus within 2 weeks after weaning. Swine practitioners attempt to optimize production by reducing the WEI. Some sows fail to resume estrous cyclicity after weaning; however, the endocrinologic pathogenesis of these anestrous sows is speculative. The average WEI is influenced by numerous factors, including season, environmental temperature, photoperiod, nutrition, stress, facility design, lactation length, and management practices. It is evident that the majority of these factors have a more profound influence on primiparous sows than on multiparous sows. Optimum protein and energy consumption by sows during lactation and after weaning and effective utilization of breeding facilities reduce the WEI. The precise roles of photoperiodic changes, elevated environmental temperatures, and stress in seasonal infertility remain poorly understood. Fortunately, current management techniques have reduced the WEI on most farms without instituting therapeutic measures.     

Hematological parameters and visceral lesions relationships in rabbit viral hemorrhagic disease.

Twenty rabbits were inoculated with a suspension of Viral Hemorrhagic Disease virus. Hemostatic functions were assessed every sixth hour from 6 to 60 hours post-inoculation. Tissue samples obtained at the same intervals allowed the study of the development of lesions throughout the experiment. Biological signs of Disseminated Intravascular Coagulation (DIC) were detected on and after 30 h post-inoculation and consisted of prolonged One Stage Prothrombin Time and Activated Partial Thrombin Time, the decrease of factors V, VII, and X and high levels of soluble fibrin monomer complexes and D-dimers. A reduction of thrombocyte numbers, heterophils and lymphocytes was associated. The close association of DIC and necrotizing hepatitis lesions suggested the hepatic lesions to be the most important DIC triggering factor. Other mechanisms are discussed.