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.     

Cystocentesis in dogs and cats]

The technique of cystocentesis in dogs and cats was studied. The indications and contraindications of this method are discussed, using illustrations.     

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.     

Genomic imprinting effects on dairy- and fitness-related traits in cattle

Data from the official milk recording scheme of the Austrian Simmental population were analysed to estimate variance components due to genomic imprinting. Traits regarded were milk yield, fat and protein content, persistency, days open (first, second and third lactation) and herd life. All dairy traits were preadjusted for BLUP herd‐year effects. After applying some data restrictions, the number of records ranged from 3391 (persistency, third lactation) to 33 993 (days open, first lactation). Two different estimation approaches were used: (i) estimation of the maternal and paternal gametic component by a dam and sire model, respectively, and (ii) estimation of the animal and an additional gametic component (maternal or paternal) by treating gametes as homozygous diploid individuals. All models also accounted for year of first calving and the cytoplasmic effect of maternal lineages. In tendency, the results of both approaches were fairly well in line with each other. For the majority of the investigated traits, no substantial imprinting effects were detected. Significant evidence of genomic imprinting on a multiple type I error ≤ 0.10 was only found for protein content and days open. For protein content in the second and third lactation, the proportions of variance due to the paternal gametic effect exceeded those of the maternal by 0.096 and 0.152, respectively. For days open in the second lactation, however, the proportion of variance of the maternal gamete effect exceeded that of the paternal by 0.036. In tendency, indication of paternal imprinting was found for all fitness‐related traits. For true and functional herd life, significant differences (pairwise type I error ≤ 0.05) of 0.040 and 0.032 were found between the proportions of variance due to the maternal and paternal gamete effect. Significant variance components of cytoplasmic effects (multiple type I error ≤ 0.01) were found for first lactation of milk yield (0.020), for first, second and third lataction of persistency (0.026, 0.035 and 0.033) and of days open (0.029, 0.016 and 0.022) and for true and functional herd life (0.019 and 0.029).     

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.     

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.     

Radiation Balance Components of Maize Hybrids Grown at Various Plant Densities

Components of the energy and heat balances were examined in two maize hybrids grown at three different plant densities (40, 70 and 100 thousand plants per hectare). One of the hybrids was drought tolerant, while the other was bred for cultivation under irrigated conditions. An increase in plant density influenced not only the size of the leaf area, but also the distribution of the leaves at various plant heights. The extinction coefficient, which provides a quantification of radiation penetration, was higher in the irrigated treatments. By contrast to the other two treatments, the plant canopy in the thinly sown stands remained open throughout the vegetation period, and thus behaved quite differently to the closed stands, making it impossible to compare them. Smaller albedo values were recorded for the hybrid bred for irrigation and in thinly sown stands. The low plant density allowed more energy to reach the soil, from which it was reflected, making a considerable contribution to the final temperature in the stand. The latent heat, in keeping with the quantity of water transpired, was the greatest in the densely sown stands. There was little difference between the latent heat values of the normal and dense stands in either hybrid, indicating that they both had a similar sensitivity to increased stand density. If sufficient water is available it would appear that the stand density could be increased even for the drought-tolerant hybrid.     

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.