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.     

Influence of Low Light Irradiance on Grain Filling in Rice (Oryza saliva L.) cultivars

Time course of grain filling pattern in two rice (Oryza sativa L.) cultivars, grown under normal and low light intensities, was studied. The number of spikelets, partially filled grains and high density grains were monitored at a 5 day interval during grain filling period from uniform panicles tagged at anthesis. The low light tolerant cultivar Swarnaprabha had more high density grains and less partially filled grains throughout the grain filling period and at harvest compared to cultivar Ratna under both normal and low light intensities. Further, the opening of spikelets in two flushes in Swarnaprabha seemed to result in a more efficient resource allocation and hence better yield as compared to Ratna, in which the opening of spikelets was in a single flush on day 10.     

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.     

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.     

Mitigation of N2O emissions from grassland by nitrification inhibitor and Actilith F2 applied with fertilizer and cattle slurry

Abstract. Nitrous oxide (N₂O) is involved in both ozone destruction and global warming. In agricultural soils it is produced by nitrification and denitrification mainly after fertilization. Nitrification inhibitors have been proposed as one of the management tools for the reduction of the potential hazards of fertilizer-derived N₂O. Addition of nitrification inhibitors to fertilizers maintains soil N in ammonium form, thereby gaseous N losses by nitrification and denitrification are less likely to occur and there is increased N utilization by the sward. We present a study aimed to evaluate the effectiveness of the nitrification inhibitor dicyandiamide (DCD) and of the slurry additive Actilith F2 on N₂O emissions following application of calcium ammonium nitrate or cattle slurry to a mixed clover/ryegrass sward in the Basque Country. The results indicate that large differences in N₂O emission occur depending on fertilizer type and the presence or absence of a nitrification inhibitor. There is considerable scope for immediate reduction of emissions by applying DCD with calcium ammonium nitrate or cattle slurry. DCD, applied at 25 kg ha^–1, reduced the amount of N lost as N₂O by 60% and 42% when applied with cattle slurry and calcium ammonium nitrate, respectively. Actilith F2 did not reduce N₂O emissions and it produced a long lasting mineralization of previously immobilized added N.     

Experimental and natural infection with the enzootic leukosis virus of cattle

A trial was performed with heifers at the age of six to seven months. The animals were experimentally infected with the lymphocytes of a virus-productive donor. Infection was produced in all the nine cases, as demonstrated by means of the positive syncytial test. As indicated by the results of the trial, the antibodies to the enzootic bovine leucosis virus (BLV) were produced soon after experimental infection. A high sensitivity of the serum-neutralization test and the ELISA method was demonstrated in this connection: by these methods, the antibodies were identified already two to three weeks after experimental infection whereas by the immunodiffusion test they could be detected only after five weeks. Twenty-four animals were exposed to natural contact infection. Within 270 days of the trial, the disease after contact was recorded only in one heifer out of the four that were in close contact with the experimentally infected animals. In this case, as compared with experimental infection, the antibodies were produced much later--after 85 to 93 days. Leucosis was recorded in none of the remaining animals. The reasons why such a favourable result was obtained were the thorough disinfection of the stables after blood collections and the strict observance of the aseptic conditions. The results of experimental infection in three cows were identical with those obtained in young cattle. In the experimentally infected dairy cows, antibodies in milk were determined by the ELISA method. As found, in milk the antibodies to BLV appear two to three weeks later than they do in serum. The ELISA method of BLV antibody detection can be used for the identification of infected animals in herds where enzootic bovine leucosis occurs.     

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.     

Pathogenesis of placentitis in the goat inoculated with Brucella abortus. II. Ultrastructural studies

Pregnant goats were inoculated intravenously or in uterine arteries with Brucella abortus, and tissues from the uterus and placenta were examined by electron microscopy. Identification of B. abortus in placentae was with antibody-coated colloidal gold. B. abortus was first seen in phagosomes of erythrophagocytic trophoblasts and in the rough endoplasmic reticulum of chorioallantoic trophoblasts. Subsequently, trophoblast necrosis and ulceration of chorioallantoic membranes were present. Coincidently, B. abortus was present in the lumen of placental capillaries. In late stages of infection, placental vasculitis was present, and placentomal trophoblasts were separated from maternal syncytial epithelium. In lesions with vasculitis, large numbers of B. abortus were in connective tissue of chorionic villi. Within the placentome, trophoblasts that lined chorionic villi contained no intracellular bacteria and were separated from B. abortus by intact basement membranes. These results suggest that bacteremic B. abortus is endocytosed by erythrophagocytic trophoblasts and that B. abortus replicates in the rough endoplasmic reticulum of chorioallantoic trophoblasts. Replication of brucellae in trophoblastic rough endoplasmic reticulum is unique; we believe that B. abortus may utilize endoplasmic reticulum for synthesis and glycosylation of bacterial membrane proteins or that B. abortus catabolizes trophoblast secretory proteins.     

Long-term application of animal slurries to grassland alters soil cation balance

Abstract. Soil samples from a 32-year grassland field experiment were taken from 0–5, 5–10, and 10–15 cm soil depths in February 2002. Plots received annual treatments of unamended control, mineral fertilizer, three rates of pig slurry and three rates of cow slurry, each with six replicates. Samples were analysed for cation exchange capacity (CEC), exchangeable cations (Na⁺, K⁺, Ca²⁺, Mg²⁺), pH and Olsen P. Exchangeable sodium percentage (ESP) was calculated as a sodicity indicator. Mean ESP was generally greater for slurry treatments than the control, with a trend of increasing ESP with application rate. This was particularly marked for cow slurry. At 0–5 cm depth ESP increased from 1.18 in the control to 1.75 at the highest rate of pig slurry and 5.60 at the highest rate of cow slurry. Similar trends were shown for CEC, exchangeable Na⁺, K⁺ and Mg²⁺, Ca²⁺ and Olsen P. The build-up of soil P due to slurry applications, together with this combination of physical and chemical factors, may increase the risk of P loss to surface waters, particularly from soils receiving high rates of cow slurry.