Selective measurement of lipoprotein lipase and hepatic triglyceride lipase in heparinized plasma from horses.

Affinity chromatography on heparin sepharose was used to identify 2 lipolytic enzymes in heparinized plasma from horses. One enzyme was typical of hepatic triglyceride lipase (HTGL), because it was resistant to inactivation by high concentrations of NaCl, and it did not require the addition of serum for activity. The other enzyme was identified as lipoprotein lipase (LPL), because of its inactivation at NaCl concentrations in excess of 0.2M, and its dependency on addition of serum as a source of apolipoprotein C-II activator. The enzymes were purified by 347-(HTGL) and 442- (LPL) fold, with yields of 54 and 58%, respectively. The partially purified enzymes were used to design incubation conditions that gave optimal activities for each enzyme in vitro. A selective assay was then developed for direct measurement of LPL and HTGL activities in heparinized plasma from horses. Analysis of HTGL took advantage of the almost complete inactivation of LPL when serum cofactor was excluded from the assay at the NaCl concentration that gave optimal HTGL activity. Prior incubation of heparinized plasma with sodium dodecyl sulfate to inhibit HTGL was necessary for measurement of LPL, because HTGL retained 67% of its activity at the NaCl concentration required for optimal LPL activity. Activity of each enzyme was measured in heparinized plasma from 12 Shetland ponies. The mean activity +/- SD for LPL was 3.22 +/- 1.04 mumol of fatty acids/ml of heparinized plasma/h (mumol of FA/ml/h. The mean activity for HTGL was 4.9 +/- 1.56 mumol of FA/ml/h. The performance of the assay was assessed by replicate analysis of pools of each enzyme with high and low activities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of 64Copper Measurements to Diagnose Canine Copper Toxicosis

Inherited canine copper toxicosis is a serious problem in Bedlington terriers and West Highland White terriers, and may also be a problem in other less-studied breeds. Affected dogs become ill at midlife with progressive and ultimately fatal liver disease. Treatments for removal of copper and prevention of copper accumulation are available, but are most effective if begun before the dog becomes ill. Until recently diagnosis has not been available until the dog is 1 year of age, and then only by an invasive liver biopsy with determination of liver copper concentration. The authors studied the use of 64copper for early diagnosis of canine copper toxicosis. Two procedures were evaluated. The first involved measuring the concentration of 64copper in blood 24 hours after oral administration of the radioisotope. At this time, 64copper was associated primarily with ceruloplasmin secreted into the blood by the liver. This procedure is useful in the diagnosis of the human counterpart, Wilson's disease. However, the authors found it to be nondiscriminatory between affected and unaffected dogs. In contrast, the second procedure, which involved measuring 64copper excreted in stool during 48 hours after an intravenous dose of radioisotope, yielded results that differentiated most affected and unaffected dogs.     

Oral fowlpox vaccination in chickens.

Chickens were given various fowlpox vaccines on food pellets--a commercial vaccine (strain M), and the same strain after a single passage on chorio-allantoic membrane or in chicken embryo fibroblasts. All three oral vaccines induced antibodies at levels similar to those induced by commercial strain M administered to the wingweb. The oral vaccine derived from chorio-allantoic membrane gave protection similar to that obtained with vaccine administered by the wingweb, but this required a thousandfold more virus.     

The effect of fatty acids in the feed on xanthine dehydrogenase activity in the Japanese quail]

The aim of the present study is to determine the effect of different fatty acids on the amino acid catabolism judged on the level of xanthine dehydrogenase (XDH). The model system of protein depletion with subsequent repletion was used in the trial consisted of Japanese quails 71 days old. In the period of protein repletion, the fatty acid (stearic, palmitic, oleic, linolenic, linoleic and lauric, respectively) was used instead of common fatty compound in the diet. Specific activities of XDH in liver, kidneys and pancreas of quails were tested dynamically from the first to the eighth day of feeding. It was demonstrated that neither lauric, palmitic nor oleic acid, respectively, had an effect on the development of the specific activity of XDH (Fig. 3). Stearic acid as a sole fatty compound in the diet however evoked reactibility of XDH in liver or in pancreas in different way as in kidneys (Fig. 4). The dynamics of the XDH development in kidneys was practically identical than that in the control. However, XDH in liver and pancreas raised statistically already from the first day of feeding. The effects of linoleic and linolenic acids on the development of specific activity of XDH were organ selective, too (Fig. 5). XDH in kidneys was shown in the same manner as in the control group. In liver and pancreas, however, the specific activity of XDH was diminished markedly just from the first day of feeding.     

Analytical methods for mycotoxins

Biological, physico-chemical and immunochemical methods are available today for solving the multiple problems which turn up in the course of mycotoxin diagnosis and mycotoxin research. In this review, selected analysis techniques are presented and their limits and possibilities are discussed. The biological methods (chicken-embryo-test, skin irritation test, cell cultures) indicate only toxicities, mostly without further specification for mycotoxins. In contrast, modern physico-chemical analysis (DC, HPLC, CLC, MS) allows the detection of defined mycotoxins in the picogram range; these methods should be employed or official and judicial purposes. The recently developed immunochemical methods (RIA, EIA) are characterized by a simple sample preparation, a short test duration and a high sensitivity; they could be used as a screening method in the course of mycotoxicological control of food and feed.     

The contribution from hyphae, roots and organic carbon constituents to the aggregation of a sandy loam under long-term clover-based and grass pastures

Water-stable macro-aggregate size fractions (>2.0 mm, 1.0–2.0 mm, 0.5–1.0 mm and 0.25–0.5 mm) and non-aggregated soil from a sandy loam under long-term clover-based pasture and from grass pasture were analysed to determine the role of acid- and water-extractable carbohydrate C, total hyphal length, microbial biomass, organic C and total and mycorrhizal root length in stabilization of the aggregates. Aggregates were examined by scanning electron microscopy (SEM) and the particle-size distribution of the size fractions was also determined. Macro-aggregation increased under grass, relative to clover-based pasture; however, the properties of the aggregate fractions measured did not reflect this difference. Microbial-biomass C, extractable-carbohydrate C, hyphal length, total and mycorrhizal root length and organic C content of the soils were poorly correlated with macro-aggregation. Within the aggregates, the proportion of 250–1000-km sand was smaller and clay, silt and fine sand (20–250 μm) were greater relative to non-aggregated soil, suggesting that the >250-μm sand in the non-aggregated soil limited the stabilization of macro-aggregates. Under SEM, no enmeshment of aggregates by hyphae and roots was apparent. Although 50–160 m hyphae g^?1 soil was found within the aggregates, calculations showed that on average only 5 to 13 lengths of hyphae were associated with each 250-μm cube of soil within the aggregates, and suggested little potential to stabilize the aggregates by enmeshing. On average, all >2.0-mm aggregates contained less than 3.6 mm of roots and less than 50% by weight of <2.0-mm aggregates contained a single length of root. The findings cast doubt about the role of hyphae and fine roots in the stabilization of macro-aggregates through an enmeshing mechanism in sandy soils.     

Production and characterization of monoclonal antibodies to budgerigar fledgling disease virus major capsid protein VP.

Eleven hybridoma cell lines producing monoclonal antibodies (MAbs) against intact budgerigar fledgling disease (BFD) virions were produced and characterized. These antibodies were selected for their ability to react with BFD virions in an enzyme-linked immunosorbent assay. Each of these antibodies was reactive in the immunofluorescent detection of BFD virus-infected cells. These antibodies immunoprecipitated intact virions and specifically recognized the major capsid protein, VP1, of the dissociated virion. The MAbs were found to preferentially recognize native BFD virus capsid protein when compared with denatured virus protein. These MAbs were capable of detecting BFD virus protein in chicken embryonated cell-culture lysates by dot-blot analysis.     

Comparative susceptibility of caponized and uncaponized tom turkeys to Pasteurella multocida.

When susceptibility to virulent Pasteurella multocida was compared, there was no significant (P greater than 0.05) difference between caponized and uncaponized tom turkeys. Neither was there any significant (P greater than 0.05) difference between the surviving caponized and uncaponized toms in the development of serum anti-P. multocida antibody. However, at 28 weeks of age, the average live body weight of the caponized toms was significantly (P less than 0.05) lower than that of the uncaponized toms. Turkeys were caponized when 9 weeks old, and different groups were exposed to P. multocida when 13, 18, 23, and 28 weeks old.