Validation of a human immunoturbidimetric assay to measure canine albumin in urine and cerebrospinal fluid.

The aim of this study was to validate an automated immunoturbidimetric assay used to quantify human albumin in urine and to accurately measure canine albumin concentrations in both urine and cerebrospinal fluid. The partial homology existing between human and canine albumin limited the accuracy of the human assays in measuring canine albumin without method modifications. Thus, the assay was modified by calibrating the analyzer with calibrators made in the laboratory containing known concentrations of canine albumin. To prepare the set of calibrators, the albumin concentration of pooled sera of healthy dogs was assessed in 5 replicates using the BromocresolGreen assay. Pooled samples were aliquoted and serially diluted to obtain the expected concentrations of albumin (0.5, 1, 5, 13, and 30 mg/dl) for establishing the canine calibration curve. Thereafter, the performance was assessed by analyzing canine urine and CSF The modified assay accurately quantified canine albumin in both specimens, as indicated by the following. Intra- and interassay variability was 0.92% and 2.74%, respectively; recovery was 99.66% and 99.07% in urine and 105.02% in CSF No interference was detected when hemolysate and glucose were added to urine. The test was linear within the verified range (0-225 mg/dl). These results demonstrate that the modified human albumin immunoturbidimetric assay can be a useful tool in the veterinary diagnostic laboratory. It is accurate and tends itself to automatization on chemistry analyzers.     

Cold tolerance during juvenile development in sorghum: a comparative analysis by genomewide association and linkage mapping

Improved cold tolerance during the juvenile phase is a major breeding goal to develop new sorghum cultivars suitable as an alternative energy crop in temperate regions. The objectives of this study were to identify marker‐trait associations for cold tolerance in a sorghum diversity panel fingerprinted with 2620 single nucleotide polymorphism (SNP) markers and to detect quantitative trait loci (QTL) in two F_2:3 populations. Traits of interest were dry matter growth rate (DMGR), leaf appearance rate (LAR), chlorophyll content (SPAD) and chlorophyll fluorescence (F_v′/F_m′ and Ф_PSII) in relation to temperature. The association panel comprised 194 genotypes, while the F_2:3 populations consisted of 80 and 92 genotypes. All populations were tested under controlled conditions in a minimum of four temperature regimes ranging from 9.4°C to 20.8°C. QTL were identified for means across environments and regression parameters describing temperature effects. Several marker‐trait associations for mean (m) DMGR, base temperature (T_b) of SPAD and Ф_PSII and temperature effect on LAR were validated by QTL detected in population 1 or 2. Promising QTL regions were found on chromosomes SBI‐01, SBI‐02, SBI‐03, SBI‐04, SBI‐06 and SBI‐09, among them genomic regions where candidate genes involved in the C‐repeat binding pathway or encoding cold‐shock proteins are located.     

Plasma levels of a low-dose constant-rate-infusion of ketamine and its effect on single and repeated nociceptive stimuli in conscious dogs

This study quantitatively investigated the analgesic action of a low-dose constant-rate-infusion (CRI) of racemic ketamine (as a 0.5 mg kg⁻¹ bolus and at a dose rate of 10 μg kg⁻¹ min⁻¹) in conscious dogs using a nociceptive withdrawal reflex (NWR) and with enantioselective measurement of plasma levels of ketamine and norketamine. Withdrawal reflexes evoked by transcutaneous single and repeated electrical stimulation (10 pulses, 5 Hz) of the digital plantar nerve were recorded from the biceps femoris muscle using surface electromyography.Ketamine did not affect NWR thresholds or the recruitment curves after a single nociceptive stimulation. Temporal summation (as evaluated by repeated stimuli) and the evoked behavioural response scores were however reduced compared to baseline demonstrating the antinociceptive activity of ketamine correlated with the peak plasma concentrations. Thereafter the plasma levels at pseudo-steady-state did not modulate temporal summation. Based on these experimental findings low-dose ketamine CRI cannot be recommended for use as a sole analgesic in the dog.     

Precision phenotyping of imidazolinone-induced chlorosis in sunflower

Chlorosis level is a useful parameter to assess imidazolinone resistance in sunflower (Helianthus annuus L.). The aim of this study was to quantify chlorosis through two different methods in sunflower plantlets treated with imazapyr. The genotypes used in this study were two inbred lines reported to be different in their resistance to imidazolinones. Chlorosis was evaluated by spectrophotometrical quantification of photosynthetic leaf pigments and by a bioinformatics-based color analysis. A protocol for pigment extraction was presented which improved pigment stability. Chlorophyll amount decreased significantly when both genotypes were treated with 10 μM of imazapyr. Leaf color was characterized using Tomato Analyzer^® color test software. A significant positive correlation between color reduction and chlorophyll concentration was found. It suggests that leaf color measurement could be an accurate method to estimate chlorosis and infer chlorophyll levels in sunflower plants. These results highlight a strong relationship between imidazolinone-induced chlorosis and variations in leaf color and in chlorophyll concentration. Both methods are quantitative, rapid, simple, and reproducible. Thus, they could be useful tools for phenotyping and screening large number of plants when breeding for imidazolinone resistance in this species.     

Production of Haploid Sugar Beet (Beta vulgaris L.) by Culturing Unpollinated Ovules

The culture of unpollinated ovules is shown to be a suitable system for the production of haploid sugar beet (Beta vulgaris L.). The yield of haploids depended upon the genotype and varied between 0 and 13 % with a mean of 1.0 %. Haploid plants could be produced from approximately 50 % of all genotypes examined. The majority of the haploids isolated (about 90%) maintained the haploid genome level during the in vitro culture and propagation; 10% of the haploid clones showed a spontaneous doubling to the diploid genome level.     

Spillover effects of a Mediterranean marine protected area on the European spiny lobster Palinurus elephas (Fabricius, 1787) resource

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Near and Mid Infrared Spectroscopy to Detect Malolactic Biotransformation of Oenococcus oeni in a Wine-Model

The aim of this work was to verify the potential of infrared （IR） spectroscopy in near and mid regions to detect the beginning of the malolactic fermentation （MLF） occurring in a model-wine and the further cells autolysis. MLF in wine is a secondary biotransformation due to lactic acid bacteria that usually occurs spontaneou,;ly or after starter inoculation at the end of alcoholic fermentation. Nowadays, it is desirable to supply winemakers with a new rapid and non-destructive approach to monitor MLF progress and 1R spectroscopy technology appears to be suitable for this purpose. The transformation of L-malic acid into L-lactic acid was carried out by inoculating a synthetic wine with an Oenococcus oeni culture and it was monitored through microbiological and chemical methods. At the same time, Fourier transform near infrared （FT-NIR） spectral data, in diffusive transflection mode using an optic probe, and FT-IR spectra, using an germanium crystal attenuated total reflectance （ATR） cell, were collected. Principal component analysis of the spectra was able to identify absorption bands related to the key molecular modifications that took place during the L-malic acid transformation. Thus, the samples were discriminated according to the fermentation phase. Although this study is a preliminary approach, results confirm that near infrared （NIR） and mid infrared （MIR） spectroscopy could be successfully applied to detect the start of MLF and the autolysis of the lactic acid bacteria （LAB） cells.     

Flavanol and bound phenolic acid contents in different barley varieties

Different barley varieties, consisting of hulled and hull-less types, of normal, waxy, and high amylose starch, as well as two-rowed and six-rowed types, were analyzed for their main proanthocyanidins and bound phenolic acids. Variations in proanthocyanidin and phenolic acid contents were studied in different barley types as well as inter-relationships between the phytochemicals and polysaccharides. The main flavanols found in the analyzed barley varieties were two dimeric as well as four trimeric forms in addition to catechin. The total amount of flavanols ranged from 325 to 527 microg/g of fresh weight of barley flour. No evident associations were found between variations in proanthocyanidin levels and different barley types. The total amount of phenolic acids ranged from 604 to 1346 microg/g of fresh weight of barley flour, with ferulic acid as the dominating acid. The amount of phenolic acids varied according to occurrence or lack of hull, with significantly higher levels in the hulled varieties.     

Associations between dry dietary factors and canine calcium oxalate uroliths

OBJECTIVE: To identify factors in dry diets associated with the occurrence of calcium oxalate (CaOx) uroliths in dogs. ANIMALS: 600 dogs with CaOx uroliths and 898 dogs without urinary tract diseases. PROCEDURE: Univariate and multivariate logistic regression were performed. RESULTS: Compared with diets with the highest concentrations of sodium, dry diets with the lowest concentrations of sodium, phosphorus, calcium, chloride, protein, magnesium, or potassium were linearly associated with increased risk of CaOx urolith formation. Significant nonlinear associations between increased occurrence of CaOx uroliths and urine acidifying potential and low moisture content were observed. Significant nonlinear associations between decreased occurrence of CaOx uroliths and carbohydrate and fiber contents were observed. A significant association between the occurrence of CaOx uroliths and dietary fat was not observed. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dry diets formulated to contain high concentrations of protein, calcium, phosphorus, magnesium, sodium, potassium, and chloride may minimize formation of CaOx uroliths. In addition, comparison of risk and protective factors of various diet ingredients fed to dogs with CaOx uroliths suggests that although similar findings were observed in canned and dry formulations, in general, greater risk is associated with dry formulations. However, before these hypotheses about dietary modifications are adopted by food manufacturers, they must be investigated by use of appropriately designed clinical studies of dogs with CaOx urolithiasis.     

Results of clinical examinations, laboratory tests, and ultrasonography in dogs with pituitary-dependent hyperadrenocorticism treated with trilostane

OBJECTIVE: To determine the efficacy of trilostane, a 3beta-hydroxysteroid dehydrogenase inhibitor, in dogs with pituitary-dependent hyperadrenocorticism (PDH). ANIMALS: 11 dogs with PDH. PROCEDURE: The initial dose of trilostane was 30 mg, PO, q 24 h for dogs that weighed < 5 kg and 60 mg, PO, q 24 h for dogs that weighed > or = 5 kg. A CBC count, serum biochemical analyses, urinalysis, ACTH stimulation test, and ultrasonographic evaluation of the adrenal glands were performed in each dog 1, 3 to 4, 6 to 7, 12 to 16, and 24 to 28 weeks after initiation of treatment. RESULTS: All dogs responded well to treatment. All had reductions in polyuria-polydipsia and panting and an increase in activity. Polyphagia decreased in 9 of 10 dogs, and 9 of 11 dogs had improvement of coat quality and skin condition. Concentration of cortisol after ACTH stimulation significantly decreased by 1 week after initiation of treatment. After treatment for 6 months, clinical signs resolved in 9 dogs. In the other 2 dogs, marked clinical improvement was reported for 1 dog, and moderate improvement was reported in the other dog. Ultrasonographically, there was a considerable change in the parenchyma and an increase in size of the adrenal glands. Adverse effects consisted of 1 dog with transient lethargy and 1 dog with anorexia. CONCLUSIONS AND CLINICAL RELEVANCE: Trilostane is an efficacious and safe medication for treatment of dogs with PDH. Additional studies in a larger group of dogs and characterization of progressive changes in adrenal glands are needed.