Dexamethasone-induced immunosuppression: a rabbit model

Rabbits are often used as animal models for experimental purposes; in many cases steroid-induced immunosuppression is necessary. The aim of this study was to characterise a model of immunosuppression in rabbits, based on changes in the lymphocyte subset distribution, changes in proliferative capacity of lymphocytes and activity of neutrophils 1, 3 and 7 days after the administration of 2mg/kg dexamethasone phosphate (DXP) three times at 6-h intervals. In peripheral blood, neutrophilia and lymphopenia together with eosinopenia, monocytopenia and basopenia in the absence of leukocytosis was detected. One day after DXP administration the absolute numbers of all lymphocyte subsets decreased in the blood, whereas in bone marrow, absolute numbers of all lymphocyte subsets increased significantly, except CD79alpha(+) cells that increased only in relative numbers. The effect of DXP on lymphocytes from the spleen, mesenteric and popliteal lymph nodes was less pronounced. In the thymus, DXP led to a marked reduction of the relative and absolute numbers of CD4(+)CD8(+) thymocytes. The proliferative capacity of lymphocytes after concanavalin A stimulation was lower in the peripheral blood and spleen only on day 1, no changes were detected in lymph nodes or in bone marrow. A marked increase in proliferative capacity was detected in the thymus. Spontaneous production of reactive oxygen metabolites by neutrophils was reduced on days 1 and 3 after DXP administration. The present results demonstrate clearly that this DXP application protocol is useful for the experimental induction of relatively short-lasting immunosuppression in rabbits.     

Interspecific germ cell transplantation: a new light in the conservation of valuable Balkan trout genetic resources?

Interspecific transplantation of germ cells from the brown trout Salmo trutta m. fario and the European grayling Thymallus thymallus into rainbow trout Oncorhynchus mykiss recipients was carried out in order to improve current practices in conservation of genetic resources of endangered salmonid species in the Balkan Peninsula. Current conservation methods mainly include in situ efforts such as the maintenance of purebred individuals in isolated streams and restocking with purebred fingerlings; however, additional ex situ strategies such as surrogate production are needed. Steps required for transplantation such as isolation of high number of viable germ cells and fluorescent labeling of germ cells which are to be transplanted have been optimized. Isolated and labeled brown trout and grayling germ cells were intraperitoneally transplanted into 3 to 5 days post hatch rainbow trout larvae. Survival of the injected larvae was comparable to the controls. Sixty days after transplantation, fluorescently labeled donor cells were detected within the recipient gonads indicating successful incorporation of germ cells (brown trout spermatogonia and oogonia—27%; grayling spermatogonia—28%; grayling oogonia—23%). PCR amplification of donor mtDNA CR fragments within the recipient gonads additionally corroborated the success of incorporation. Overall, the transplantation method demonstrated in this study presents the first step and a possible onset of the application of the germ cell transplantation technology in conservation and revitalization of genetic resources of endangered and endemic species or populations of salmonid fish and thus give rise to new or improved management strategies for such species.     

Immobilization of cell wall invertase modified with glutaraldehyde for continuous production of invert sugar

Yeast cell wall invertase (CWI) was modified with dimethyl suberimidate, glutaraldehyde, formaldehyde, and sodium periodate. Retained activity after modification was 45% for CWI modified with formaldehyde, 77% for CWI modified with sodium periodate, 80% for CWI modified with glutaraldehyde, and 115% for CWI modified with dimethyl suberimidate. Chemically modified and native CWIs showed significantly broad pH stability (pH 3-11), whereas after incubations at 50, 60, and 70 °C, CWI modified with glutaraldehyde showed the highest thermostability. Optimum pH for CWI modified with glutaraldehyde was between 4 and 5, whereas optimum temperature was at 60 °C. Comparison to CWI modified with glutaraldehyde after immobilization within alginate beads showed broader pH optimum (4.0-5.5) as well as broader temperature optimum (55-70 °C). Column bed reactor packed with the immobilized CWI modified with glutaraldehyde was successfully used for the 95% inversion of 60% (w/w) sucrose at the flow rate of 3 bed volumes per hour, pH 4.9, and 45 °C. A 1 month productivity of 3844 kg of inverted sugar/kg of the immobilisate was obtained.     

Nondestructive evaluation of anthocyanins in olive (Olea europaea) fruits by in situ chlorophyll fluorescence spectroscopy

Anthocyanins (Anths) in olive (Olea europaea L.) fruits at different degrees of pigmentation were assessed nondestructively by measuring chlorophyll fluorescence (ChlF). The method is based on the comparison of the ChlF excitation spectra from olives with different pigmentation from green to green-red, reddish-purple, and purple. The logarithm of the ratio between the fluorescence excitation spectra (logFER) from two different colored zones gave the difference in the absorption spectrum between them. The absorbance spectrum derived from the logFER between a red olive and the same olive devoid of the skin showed the typical Anth green band (at 550 nm). It matched that recorded by microspectrophotometry on a single pulp cell and the in vitro absorbance spectrum of the olive skin extract. As expected, the in vivo Anths absorption maximum increased in intensity going from less to more mature olives and was higher in the sun-exposed olive side with respect to the sun-shaded side. Absolute quantitative nondestructive determination of Anths for each olive sample was obtained by the logFER calculated for two excitation wavelengths, 550 and 625 nm, of ChlF at 740 nm. Going from green to purple skin colors, the Log[ChlF(625)/ChlF(550)] was fairly well-correlated to the extract Anths concentration. Finally, the relationship between the Anths and the other main phenolics present in the olives analyzed by high-performance liquid chromatography was evaluated. The main result was a net increase of verbascoside with increasing Anths content. On the basis of our results, the development of a new rapid and noninvasive method for the monitoring of olive development and ripening can be envisaged.     

Effect of different management practices on water quality of intensive tilapia culture systems in Israel

Commercial intensive aquaculture systems werebuilt and are managed in a somewhat differentway in each farm. To evaluate the effects ofseveral management procedures on water qualityin intensive fish ponds, data from severallocations, times and culture conditions indifferent farms were collected and are hereinanalyzed through multivariate statistics.Water quality in the intensive ponds depends onthe water entering, the biological processeswithin, and the water leaving the ponds. Areservoir used as source and sink water supplied theintensive ponds with higher organic loadingthan clear source waters, and its phytoplanktoncontent affected nitrogen cycling within theintensive ponds. The systems with a reservoirhad better water quality in the intensive pondsthan those with only clean source water.Within the ponds (1) compared to paddle-wheelaeration, aeration by pure oxygen increasedoxygen concentration, improved nitrificationand promoted decomposition that reduced organicloading. (2) In concrete ponds accumulation oforganic matter and development of anerobicconditions on the pond bottom was higher thanin the slippery plastic-covered ponds. (3) Allintensive ponds provided good growthconditions, tilapia biomass having relativelysmall influence on water quality. Only inpaddle-wheel aerated ponds did increased tilapiabiomass increased inorganic nitrogen compoundsand soluble phosphorus through excretion, andreduce organic nitrogen through a moreefficient removal of food particles.Water leaving the ponds removes matteraffecting water quality within the pond. (1)Draining sediments accumulated on the bottomavoided development of anaerobic conditionswhere denitrification and phosphorus liberationcan occur. (2) Water exchange removed particleswith nitrifying bacteria and algae that absorbnutrients. A high water exchange rate may havea negative effect from the water quality pointof view and from the extra costs incurred inenergy and feeds washed out.The processes described occur simultaneouslythroughout the culture period and shape waterquality dynamics in the ponds. This researchcontributed to the understanding of howmanagement procedures affect the differentphases of water quality dynamics in real-scaletilapia commercial intensive systems.     

Effects of dietary protein content on renal parameters in normal cats

This study evaluates the effect of dietary protein content on renal parameters in 23 healthy spayed female cats. The objective was to determine if cats eating diets high in protein will have higher serum urea nitrogen (UN) and creatinine values without a detectable change in kidney function, as assessed by urinalysis. A single random cross-over design was used. Cats were fed a standard maintenance diet for at least 1 month prior to the dietary trial. They were fed in two phases. For the first phase, cats were randomly assigned to receive either a high protein [HP=46% metabolizable energy (ME)] or low protein (LP=26% ME) diet. For the second phase, cats were fed whichever diet they were not fed during the phase I period. Blood and urine samples were collected at 2-week intervals for the duration of the study (10 weeks). UN, albumin, alanine aminotransferase and urine specific gravity were significantly higher, and creatinine and phosphorus were significantly lower (P<0.05) when cats were fed the HP diet as compared to when they were fed the LP diet, although none of the mean values were found to be outside of the corresponding reference interval. Dietary intake can result in clinically significant changes in UN and statistically significantly changes in several other biochemical analytes, although all analytes are likely to remain within normal reference intervals. Therefore, an accurate dietary history is necessary to help determine if renal parameters are being influenced by diet in a particular patient.     

Gastroduodenoscopy in the dog and cat.

Endoscopic examination of the upper gastrointestinal (GI) tract in dogs and cats is a relatively recent development. It was just over 20 years ago, in 1978, that the first series report of GI endoscopy in dogs and cats was presented. In those initial days, endoscopy was truly a novelty, and the primary uses for endoscopes were to retrieve foreign objects or visualize the esophagus and stomach. It was not until the mid-1980s that technology and training made gastroduodenoscopy standard practice. The evolution in endoscopic technology has also mirrored the evolution in computers and imaging modalities. The original fiberoptic endoscopes, although still available, are giving way to video endoscopy and digital imaging capabilities that provide spectacular images and an ability to generate hard copy images for medical records, teaching purposes, and research data that was previously not possible.     

A Data Generator for Evaluating Spatial Issues in Precision Agriculture

With the rapid rise in site-specific data collection, many research efforts have been directed towards finding optimal sampling and analysis procedures. However, the absence of widely available high quality precision agriculture data sets makes it difficult to compare results from separate experiments and to assess the optimality and applicability of procedures. To provide a tool for spatial data experimentation, we have developed a spatial data generator that allows users to produce data layers with given spatial properties and a response variable (e.g. crop yield) dependent upon user specified functions. Differences in response functions within fields can be simulated by assigning different models to regions in coordinate-(x and y) or feature space (multidimensional space of attributes that may have an influence on response). Noise, either unexplained variance or sensor error, can be added to all spatial layers. Sampling and interpolation error is modeled by sampling a continuous data layer and interpolating values at unsampled locations. The program has been successfully tested for up to 15000 grid points, 10 features and 5 models. As an illustration of the potential uses of generated data, the effect of sampling density and kriging interpolation on neural network prediction of crop yield was assessed. Yield prediction accuracy was highly related (correlation coefficient 0.98) to the accuracy of the interpolated layers indicating that unless data are sampled at very high densities relative to their geostatistical properties, one should not attempt to build highly accurate regression functions using interpolated data. By allowing users to generate large amounts of data with controlled complexity and features, the spatial data generator should facilitate the development of improved sampling and analysis procedures for spatial data.