A rabbit model of non-typhoidal Salmonella bacteremia

Bacteremia is an important cause of morbidity and mortality in humans. In this study, we focused on the development of an animal model of bacteremia induced by non-typhoidal Salmonella. New Zealand White rabbits were inoculated with a human isolate of non-typhoidal Salmonella strain CVD J73 via the intra-peritoneal route. Blood samples were collected at specific time points and at euthanasia from infected rabbits. Additionally, tissue samples from the heart, lungs, spleen, gastrointestinal tract, liver and kidneys were obtained at euthanasia. All experimentally infected rabbits displayed clinical signs of disease (fever, dehydration, weight loss and lethargy). Tissues collected at necropsy from the animals exhibited histopathological changes indicative of bacteremia. Non-typhoidal Salmonella bacteria were detected in the blood and tissue samples of infected rabbits by microbiological culture and real-time PCR assays. The development of this animal model of bacteremia could prove to be a useful tool for studying how non-typhoidal Salmonella infections disseminate and spread in humans.     

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.     

Serum amyloid A isoforms in serum and milk from cows with Staphylococcus aureus subclinical mastitis

Serum amyloid A proteins (SAA) are very sensitive acute phase proteins, displaying multiple isoforms in plasma and different body fluids. They are currently under investigation as biomarkers of diseases. The aim of the present study was to compare the concentration and isoform expression of SAA in serum and milk of cows with bacteriologically negative milk (control group) and naturally occurring Staphylococcus aureus (S. aureus) subclinical mastitis (subclinical mastitis group). Somatic cell count (SCC) and bacteriological analyses were performed to establish the control and subclinical mastitis group. SAA concentration was evaluated using a commercial ELISA kit, while expression of different isoforms (serum A-SAA and milk M-SAA3 isoforms) was visualized by denaturing isoelectrical focusing and immunoblotting. The SAA concentrations in sera and milk of cows in the subclinical mastitis group were three and 100 times higher than in those from the control group of cows, respectively. Cows in the subclinical mastitis group had more acidic SAA isoforms in serum with the most prominent one at pI 5.5. This isoform was not detected in sera from the control group. Milk samples in the subclinical mastitis group contained abundant highly alkaline M-SAA3 isoforms and most of the serum isoforms, except for that at pI 5.5. In the subclinical mastitis group SAA isoforms with equivalent pI as serum isoforms accounted for 20% of the total SAA concentration in milk. There were significant differences in the concentrations and isoform patterns of SAA in serum and milk between the control and subclinical mastitis groups of cows. Also, we demonstrated that serum SAA isoforms were not transferred to milk proportion to their plasma content.     

Pancreatitis in cats: diagnosis and management of a challenging disease

Feline pancreatitis can be a very difficult disease to diagnose and often requires a combination of clinical suspicion, appropriate physical examination findings, elevations in serum feline pancreatic lipase immunoreactivity, and changes on abdominal ultrasonography consistent with pancreatic disease. The diagnostic difficulties encountered are related to a lack of specific and readily attributable clinical signs in cats. The sensitivity and specificity of the diagnosis of pancreatitis are highest when a combination of tests is utilized; but even when such tests are employed, the diagnosis is still problematic, especially in cats with chronic pancreatitis. Therapy is symptomatic and focuses on maintaining fluid volume, controlling pain and vomiting, preventing infection, and adjusting to changes in the cat's condition as they occur.     

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.     

Relationship between soil CO2 concentrations and forest-floor CO2 effluxes

To better understand the biotic and abiotic factors that control soil CO₂ efflux, we compared seasonal and diurnal variations in simultaneously measured forest-floor CO₂ effluxes and soil CO₂ concentration profiles in a 54-year-old Douglas fir forest on the east coast of Vancouver Island. We used small solid-state infrared CO₂ sensors for long-term continuous real-time measurement of CO₂ concentrations at different depths, and measured half-hourly soil CO₂ effluxes with an automated non-steady-state chamber. We describe a simple steady-state method to measure CO₂ diffusivity in undisturbed soil cores. The method accounts for the CO₂ production in the soil and uses an analytical solution to the diffusion equation. The diffusivity was related to air-filled porosity by a power law function, which was independent of soil depth. CO₂ concentration at all depths increased with increase in soil temperature, likely due to a rise in CO₂ production, and with increase in soil water content due to decreased diffusivity or increased CO₂ production or both. It also increased with soil depth reaching almost 10 mmol mol⁻¹ at the 50-cm depth. Annually, soil CO₂ efflux was best described by an exponential function of soil temperature at the 5-cm depth, with the reference efflux at 10 °C (F₁₀) of 2.6 μmol m⁻² s⁻¹ and the Q₁₀ of 3.7. No evidence of displacement of CO₂-rich soil air with rain was observed.Effluxes calculated from soil CO₂ concentration gradients near the surface closely agreed with the measured effluxes. Calculations indicated that more than 75% of the soil CO₂ efflux originated in the top 20 cm soil. Calculated CO₂ production varied with soil temperature, soil water content and season, and when scaled to 10 °C also showed some diurnal variation. Soil CO₂ efflux and concentrations as well as soil temperature at the 5-cm depth varied in phase. Changes in CO₂ storage in the 0–50 cm soil layer were an order of magnitude smaller than measured effluxes. Soil CO₂ efflux was proportional to CO₂ concentration at the 50-cm depth with the slope determined by soil water content, which was consistent with a simple steady-state analytical model of diffusive transport of CO₂ in the soil. The latter proved successful in calculating effluxes during 2004.     

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.     

Preservation of common carp germ cells under hypothermic conditions: Whole tissue vs isolated cells

The aim of this study was to optimize the conditions for hypothermic storage of spermatogonial stem cells (SSCs) and oogonial stem cells (OSCs) of common carp Cyprinus carpio. This was conducted by storing gonadal tissue or isolated cells for 24 hr under hypothermic conditions in the first experiment and by testing two different storage media (L‐15 or DMEM supplemented with 10% FBS and 25 mM HEPES) and regular medium change (every 4 days) during two weeks of hypothermic storage in the second experiment. During the first 24 hr, isolated cells showed no decrease in viability, while cells obtained from hypothermically stored tissues displayed significantly lower viability after only 6 hr (Tukey's HSD, p < 0.01) indicating that hypothermic storage of isolated cells is superior to storing tissue pieces. The 2‐week trial demonstrated that storage media have a profound influence, while regular medium exchange does not have a positive effect on cell viability. Viability of SSCs and OSCs after two weeks was approximately 40% and 25%, respectively; however, survival of ~70% was obtained after 10 days of storage for SSCs and 7 days for OSCs. Hypothermic storage developed in this study has many practical applications during the development of surrogate broodstock technologies for common carp, but also in carp hatcheries and for the conservation of genetic resources of closely related cyprinid species.     

Measurement of Resting and Stress-elevated Serum Cortisol in Rainbow Trout Oncorhynchus mykiss in Experimental Net-pens

A commercially available heterogeneous, solid-phase tube enzyme-linked immunoassay (ELISA) was modified and validated for the measurement of serum cortisol in rainbow trout Oncorhynchus mykiss . The assay is accurate and precise. Resting and stress-elevated serum cortisol concentrations were measured in rainbow trout with a sensitivity of 1.5 ng/ml. Fish held in net-pens at a density of 0.4 kg/m³/cm had a resting cortisol level of 16.5 ± 3.8 ng/ml (mean ± SE). At 3 h postdisturbance, serum cortisol levels were not affected by the removal of fish from adjacent net-pens with dip nets or by the use of 200 mg/L tricaine methanesulfonate (MS-222) as an anesthetic for obtaining samples. However, an acute stress (60 s removal from water) elevated serum cortisol levels to 73.7 ± 9.4 ng/ml.