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.     

The Effect of Diet on the Biochemical Composition of Juvenile Artemia: Potential Formulations for Rock Lobster Aquaculture

The lipid class and fatty acid (FA) composition of juvenile Artemia fed continuously on four diets—the microalga Tetraselmis suecica , a mix of oat bran-wheat germ-lecithin (OWL), OwL-eicosapentaenoic acid (EPA), and OWL-EPA-arachidonic acid (AA)—were examined over a 9-d experiment in an attempt to approximate the FA profile of phyllosoma larvae of wild southern rock lobster Jasus edwardrii . The main difference in lipid class composition of Artemia fed the four diets was the relative level of polar lipid (PL) and triacylglycerol (TAG). By day 9, the algal-fed Artemia were highest in PL (95% of total lipid) and lowest in TAG (2%), whereas the remaining diets resulted in Artemia with 16–30% PL and 41–82% TAG. After 2 d, the relative FA composition of all Artemia treatments closely reflected those of the diets, with no marked change after further feeding (to day 9). In terms of the content of essential polyunsaturated fatty acids (PUFA), by day 5 Artemia fed: 1) with the algal diet contained 7 mg/g FA dry mass (0.3% DHA, 6.3% EPA, 3.4% AA of total FA); 2) with the OWL diet contained 3 mg/g (0.3% DHA, 0.9% EPA, 0.7% AA); 3) with the OWL-EPA diet contained 55 mg/g (6.2% DHA, 11.6% EPA, 1.1% AA); and 4) with the OWL-EPA-AA contained 83 mg/g (3.8% DHA, 7.5% EPA, 17.4% AA). The PUFA profiles of Artemia using the OWL-oil diets were similar to wild rock lobster phyllmmata, although levels of doco-sahexaenoic acid (DHA) were lower (10% DHA) than in J. edwardsii larvae. On the basis of PUFA composition data alone, the results suggest the suitability of the OWL-oil mixed diets for consideration for feeding to Artemia used in the culture of southern rock lobster larvae, particularly if the level of DHA can be further enhanced.     

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.     

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.     

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.     

Blastocyst Evaluation by Means of Differential Staining: a Practical Approach

Techniques for in vitro production of embryos have been developed world-wide in different species, with promising results in human and ruminants. Thousands of human IVF-babies have been born during the last 20 years and thousands of in vitro -produced calves have been born since the late 1980s. With current methods for bovine in vitro fertilization, about 30–40% of in vitro -fertilized bovine oocytes develop further to the blastocyst stage and can be used for transfer. A proper evaluation of blastocyst quality remains however, an important challenge for every researcher involved in embryology and for every clinician who wants to select the best embryos for transfer. This review attempts to summarize the different methods available for estimation of blastocyst quality with a special emphasis upon differential staining.