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.     

Insulin and glucose regulation.

Abnormally high or low blood glucose and insulin concentrations after standardized glucose tolerance tests can reflect disorders such as pituitary dysfunction, polysaccharide storage myopathies, and other clinical disorders. Glucose and insulin responses, however, are modified by the diet to which the animal has adapted, time since it was last fed, and what it was fed. Body fat (obesity), fitness level, physiologic status, and stress also alter glucose and insulin metabolism. Therefore, it is important to consider these factors when evaluating glucose and insulin tests, especially if only one sample it taken. This article describes the factors affecting glucose and insulin metabolism in horses and how they might influence the interpretation of standardized tests of glucose tolerance.     

The Role of Hepatic Ultrasonography in Small Animal Medicine: From the Identification of the Lesions to Diagnosis

Veterinary Research Communications -     

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.     

Detection of feline coronavirus infection in captive cheetahs (Acinonyx jubatus) by polymerase chain reaction.

Feline coronavirus genetic elements were detected by polymerase chain reaction from blood, fecal samples, and effusive fluid collected from 33 cheetahs in the U.S.A. Feline coronavirus-specific serum antibodies were also measured by indirect immunofluorescence. Ten cheetahs were positive for viral shedding by polymerase chain reaction, whereas 13 were seropositive by immunofluorescence. Results of serology did not consistently correlate with shedding of virus, and the capture antigen used for detection of feline coronavirus-specific antibodies had a significant impact on results. Testing of samples from one population over a 1-yr period indicated chronic infection in some animals. These relatively healthy carrier animals were a source of virus for contact animals. Screening programs in cheetah populations for feline coronavirus infection may be most reliable if a combination of serologic analysis and viral detection by polymerase chain reaction is used.     

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.