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.     

Antimicrobial therapy for gastrointestinal diseases.

Antibiotics will always be needed in horses for many types of infections, but the adverse consequences also must be considered. For the conditions described in this article, there is justification for antibiotic therapy. The intestinal problems that antibiotics can induce are among the risks from their administration to horses. Disruption of the endogenous bacterial population, colitis, and diarrhea are the most common complications from antibiotic therapy.     

Using 3D animations to teach intracellular signal transduction mechanisms: taking the arrows out of cells

Traditional methods of teaching intracellular biological processes and pathways use figures or flowcharts with the names of molecules linked with arrows. Many veterinary students, presented with such material, simply memorize the names or chemical structures of the molecules and are then likely to forget the material once the examination is completed. To address this problem, the authors designed, created, and field-tested new teaching media that incorporate realistic three-dimensional (3D) animations depicting the dynamic changes that occur in intracellular molecules during cellular activation. Testing found that veterinary students taught using traditional teaching media (e.g., lectures, handouts, textbooks) are proficient in memorizing the names and order of intracellular molecules but unable to appreciate the interactions between these elements or their spatial relationships within cells. In contrast, more than 90% of veterinary students taught using 3D animations not only recall the facts about the intracellular elements but also develop accurate mental images of the interactions among these molecules and their spatial relationships. These findings strongly suggest that the comprehension of complex biological processes by veterinary students can be enhanced by the use of dynamic 3D depictions of these processes in the classroom.     

Effects of bacterial lipopolysaccharide on thermoregulation in green anole lizards (Anolis carolinensis)

Fever is a non-specific host defense mechanism that comprises part of the innate immune response. Innate immune function is thought to be an important adaptive immunological response to infection because it occurs across a broad diversity of phyla. Some reptiles can mount a febrile response, despite the fact that their internal body temperatures (T(b)s) are, to some extent, controlled by the environmental temperatures in which they live. This study was undertaken to determine if LPS would induce fever in green anole lizards (Anolis carolinensis). Lizards were maintained in thermal gradients (22-45 degrees C) with a 12-h diurnal cycle. anoles were injected with LPS, pyrogen-free saline, or left untreated, and their T(b)s were recorded every 15min using internal cloacal probes. All lizards showed a diurnal periodicity in T(b) characterized by decreased temperatures during the scotophase (dark hours) and higher temperatures during the photophase (light phase). Anoles injected with LPS exhibited a hypothermic response, relative to untreated and saline-injected animals. The response varied from 2.1 to 4.6 degrees C lower than control lizards. The hypothermic response was initiated within 12-24h of LPS injection, and continued for 3 days after treatment. However, the anapyrexic response was observed primarily during scotophases, with photophase hypothermia observed only on the first day after LPS injection.     

Reproducibility of bovine neonatal pancytopenia (BNP) via the application of colostrum

A haemorrhagic diathesis has been observed in young calves since 2007 which is described as bovine neonatal pancytopenia (BNP) and presents a completely new disease. The objectives of our investigation were to test if BNP could be reproduced using colostrum of cows with a BNP history and pre-colostral calves from farms where BNP has not been observed. In the present experiment, 22 German Holstein calves from BNP-free farms were fed four to six hours after birth 2.5 l colostrum from cows which had been reported to have had at least one calf with BNP in the last lactation. We distinguished three different experimental groups according to the composition of the colostrum. In experimental group I, each of the six calves received colostrum of a single cow, in experimental group II all six calves received colostrum from the same cow and in experimental group III each of the ten calves received a colostrum mix from ten different cows. Clinical signs of BNP were observed in 50% of the calves in experimental group I, 67% of the calves in experimental group II and all calves in experimental group III. The lethality in the three experimental groups was significantly different with rates of 16.7%, 66.7% and 80%, respectively. Calves fed with a colostrum-mix in experimental group III had the highest lethality. Neither the farm nor the amount of the colostrum fed had a significant effect on the occurrence and course of BNP. The profiles for thrombocytes, leucocytes and erythrocytes significantly differed in dependence of the severity of BNP signs. Calves with non-lethal BNP showed thrombocytopenia with values below 100 G/l on the 1th to 3rd and the 7th to 11th day of life. In calves with lethal BNP, thrombocytes decreased under 50 G/l from day 5. In calves with non-lethal BNP, a decrease of the leucocytes under the threshold was present only for a short period of time. In calves with lethal BNP, leucocytes decreased in the first 5 days after birth continuously and increased on the 6th to the 8th day to normal values and then a rapid decrease occurred. Erythrocytes decreased under the normal threshold just in the last two days before the calves died or were euthanized. Thus, the present experiments showed that colostrum of cows with a BNP-history and vaccination with PregSure BVD from Pfizer caused lethal BNP. We can assume that the different reactions of the calves are due to immunogenetic reactions to colostral alloreactive antibodies. The reaction spectrum of calves depends on the presence of antigens which can react with these colostral antibodies. The experimental results can explain the different incidences of BNP within and among farms as well as between breeds.