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)     

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.     

The internet and the future of equine practice.

The Internet is a fast-growing technology that allows information (text as well as images) to be shared across the world in an instant. As computer systems, software, and Internet services become less expensive options, the world becomes more experienced and comfortable with these technologies. In the foreseeable future, equine veterinarians are likely to be better educated about diseases and treatments, better connected with specialists for consultations on difficult cases, able to respond to client communications more efficiently, able to accurately educate clients, and able to communicate with colleagues from across the country and the world. Additionally, there is likely to be a time when using the Internet and computers is profitable in all these ways as well as economically. Improving doctor-client relationships and patient care are goals for all veterinarians, and the Internet holds the technology to render these now and in the future.     

Investigation of Cervical Patency and Uterine Appearance in Domestic Cats by Fluoroscopy and Scintigraphy

The cervical patency of six domestic female cats was monitored under sedation by infusion of contrast medium (Omnipaque) into the cranial vagina during early oestrus, mid‐oestrus, late oestrus and interoestrus or a radiopharmaceutical (^99mTc‐HSA) during mid‐ and interoestrus in a non‐ovulatory oestrous cycle. The transport of the contrast medium or the radiopharmaceutical through the cervix and within the uterine horns was observed under fluoroscopy and with the aid of scintigraphy. In three of the queens, transcervical transport of contrast medium was demonstrated in all stages of oestrus, in one queen during mid‐oestrus, late oestrus and 1 day after oestrus, and in two queens only during late oestrus. The relations between the cervical patency to the contrast medium and the oestrous behaviour, cornification of the vaginal cells and the serum oestradiol‐17β concentration were evaluated, and a relationship was found between the cervical patency and the degree of vaginal cornification. Transcervical transport of the radiopharmaceutical was observed in three queens during mid‐oestrus. When the cervix was open, hysterography under a fluoroscope and hysteroscintigraphy were performed. The fluoroscopic and scintigraphic recordings revealed the patterns of the uterine contractions during oestrus in both ascending and descending directions, and the movement of the uterine contents back and forth between the uterine horns. The hysterograms were classified according to the shape of the uterine horns and the appearance of the endometrial lining. Spiral‐shaped uterine horns with a smooth inner contour were observed in two queens, and a corkscrew appearance with irregular filling defects in the uterine lumen was shown in two queens that had developed subclinical cystic endometrial hyperplasia. These findings demonstrated that fluids or particles deposited in the cranial vagina of the cat can be transported into the uterus during some stages of the oestrous cycle. The fluoroscopic and scintigraphic techniques developed in this study may be further modified to permit more detailed studies of uterine contractile patterns and sperm transport in the feline female reproductive tract. Hysterography proved useful to diagnose uterine disease. The information on cervical patency is of value also for the development of techniques for artificial insemination in this species, and should be studied also in the ovulatory cycle.     

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.     

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.     

Long-term application of animal slurries to grassland alters soil cation balance

Abstract. Soil samples from a 32-year grassland field experiment were taken from 0–5, 5–10, and 10–15 cm soil depths in February 2002. Plots received annual treatments of unamended control, mineral fertilizer, three rates of pig slurry and three rates of cow slurry, each with six replicates. Samples were analysed for cation exchange capacity (CEC), exchangeable cations (Na⁺, K⁺, Ca²⁺, Mg²⁺), pH and Olsen P. Exchangeable sodium percentage (ESP) was calculated as a sodicity indicator. Mean ESP was generally greater for slurry treatments than the control, with a trend of increasing ESP with application rate. This was particularly marked for cow slurry. At 0–5 cm depth ESP increased from 1.18 in the control to 1.75 at the highest rate of pig slurry and 5.60 at the highest rate of cow slurry. Similar trends were shown for CEC, exchangeable Na⁺, K⁺ and Mg²⁺, Ca²⁺ and Olsen P. The build-up of soil P due to slurry applications, together with this combination of physical and chemical factors, may increase the risk of P loss to surface waters, particularly from soils receiving high rates of cow slurry.