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.     

Enhanced recovery of avian influenza virus isolates by a combination of chicken embryo inoculation methods.

Since 1993, 14 cases of avian influenza from four different states in the U.S.A. have been diagnosed by virus isolation from eight avian species. Only 11 of the 14 avian influenza virus (AIV) primary isolations would have been successful if only the standard protocol for AIV isolation, i.e., inoculation of specific-pathogen-free embryonating chicken eggs (ECEs) by the chorioallantoic sac (CAS) route, had been followed. Primary isolation attempts were negative for AIV in three cases in which ECEs were inoculated by the CAS route; AIV could not be detected by hemagglutinating activity, agar gel immunodiffusion test or negative stain electron microscopy. However, in these three cases, primary isolations of AIV were achieved by inoculation of ECEs into either the yolk sac or onto the chorioallantoic membrane.     

Chemopreventive efficacy of lycopene on 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis.

The chemopreventive efficacy of lycopene on 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis was examined using lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR) as biomarkers of chemoprevention. Twenty four male Syrian hamsters were divided into four groups of six animals each. The right buccal pouches of the animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin three times a week. The animals in group 2 were painted with DMBA as in group 1 and in addition received 2.5 mg/kg body weight lycopene orally three times a week on days alternate to DMBA application. Group 3 animals received lycopene as in group 2. Animals in group 4 received neither DMBA nor lycopene and served as control. The hamsters were killed after an experimental period of 14 weeks. Biochemical measurements were carried out in tumour and normal tissues. All hamsters painted with DMBA alone for 14 weeks developed well-differentiated squamous cell carcinomas. Diminished lipid peroxidation in the oral tumour tissue was accompanied by a significant increase in the levels of GSH, GPx, GST and GR. Administration of lycopene significantly suppressed DMBA-induced oral carcinogenesis as revealed by the absence of carcinomas. The results of the present study suggest that lycopene may exert its chemopreventive effects by modulating lipid peroxidation and enhancing the activities of the enzymes in the glutathione redox cycle.     

Reproductive roles predict sexual dimorphism in internal and external morphology of lake whitefish,Coregonus clupeaformis

Abstract – The different reproductive roles of the sexes can predict the direction and magnitude of sexual dimorphism of external and internal morphology. Males should have enlarged structures that enhance the acquisition of mating opportunities, whereas females are predicted to have enlarged organs that are associated with the production of eggs. We tested these predictions in male and female lake whitefish, a species in which both sexes have similar overall body size and shape. After controlling for body size, male lake whitefish had significantly longer jaws and pectoral and pelvic fins, larger hearts, and more muscle than females. Sexual dimorphism in relative muscle mass may be one of the most fundamental morphological differences between males and females. Females had relatively heavier livers than males. Because the liver is important for the breakdown of fats and vitellogenesis, selection should favour an enlarged liver in females for the processing of energy and the production of large numbers of eggs.     

P‐20 Clinical,cytopathological and histopathological evaluation of sporotrichosis in experimentally infected cats

The purpose of this study was to evaluate the clinicopathological aspects of experimental sporotrichosis in cats and compare the sensitivity of cytopathology, histopathology and culture as diagnostic tools in different phases of the infection. Twenty adult, mixed‐breed cats (10 males and 10 females) were inoculated subcutaneously with 10⁶ fungal microorganisms. Clinical examination was performed weekly. Cytopathologic, histopathologic and culture examinations were performed at 15, 30 and 60 days postinoculation. Culture of multiple organs was performed after euthanasia at 30 (10 cats) and 60 (10 cats) days postinoculation. Friedman parametric and nonparametric statistical analysis were applied to the results. The nodular, tumoral and necrotic lesionsprogressed significantly until day 30 postinoculation, and partial spontaneous regression occurred at day 60, particularly in males. An intense inflammatory pyogranulatomous and lymphocytic infiltrate with rare giant cells and sparse fibrosis associated with numerous, pleomorphic, intra‐ and extracellular fungal cells were observed on day 30. These findings gradually decreased by day 60. Despite the inflammatory granuloma associated with feline sporotrichosis, a tendency for dissemination was observed, with fungal isolation in the lymph nodes, spleen and liver at the 30 and 60 days postinoculation. No significant differences were observed between cytopathology, histopathology and fungal culture during the different phases of the disease. Therefore, cytological examination was considered a simple, rapid and inexpensive diagnostic method at all stages of this disease. Funding: Self‐funded.     

Evaluation of Five Most Commonly Grown Cotton Cultivars (Gossypium hirsutum L.) Under Mediterranean Conditions: Productivity and Fibre Quality

Studies were conducted in four regions of central Greece (Larisa, Karditsa, Livadia, Thiva), to evaluate the productivity and fibre properties (fibre strength and fibre length) of the five most commonly grown cotton cultivars in these regions. The experiments lasted for 6 years (1995–2000). All the characteristics tested were strongly influenced by the factors ‘year’, ‘cultivar’ and ‘region’. Significant cultivar × region, year × cultivar, region × year and cultivar × region × year interactions occurred for all the characteristics studied. The cultivar Alegria had the highest seed‐cotton and lint yield, while Aria had the lowest ones. Both cultivars gave the highest mean lint percentage but the lowest mean boll weight (MBW). Alegria had the lowest fibre strength, while Aria had the highest. The Acala‐type cultivars, Acala SJ2 and Zeta 2, had intermediate seed‐cotton and lint yield, while Vered 171 had high seed‐cotton and satisfactory lint yield. The three cultivars mentioned above had the highest MBW and very good fibre strength. Positive correlation was observed between fibre strength and length. In spite of the fact that mean fibre length was statistically the same in all cultivars, it was influenced by the factor ‘year’, as a result of different weather conditions. The year 1996 was the most unfavourable one for seed‐cotton yield and MBW, because it was characterized by lower than average air temperatures and rather dry conditions for all study sites. The factor ‘region’, a combination of different climatic conditions and cultural practices, influenced significantly all the parameters studied.     

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.