Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

Oral administration of tepoxalin in the horse: a PK/PD study

Tepoxalin is a non-steroidal anti-inflammatory drug with analgesic, anti-inflammatory, and antipyretic properties and has been recently introduced into veterinary medicine. The aim of this study was to evaluate the pharmacokinetic/pharmacodynamic (PK/PD) profile of tepoxalin to assess whether it would be suitable for clinical use in horses. Six female fasting/fed horses were given 10mg/kg tepoxalin orally in a cross-over study. After administration, tepoxalin underwent rapid and extensive hydrolytic conversion to its carboxylic acid metabolite RWJ-20142. In animals that had been fed, the plasma concentrations of tepoxalin were undetectable, whereas in fasting animals they were close to the limit of quantification of the method. No differences between the fasting/fed groups in RWJ-20142 plasma concentrations were shown. Tepoxalin showed a strong and long-lasting ex vivo inhibitory activity against cyclooxygenase (COX)-1, mainly due to its main metabolite RWJ-20142. Tepoxalin and RWJ-20142 do not seem to possess either COX-2 or 5-lipoxygenase inhibitory activity in the horse. These features suggest that the drug is a selective COX-1 inhibitor in horses, with no significant anti-inflammatory activity. Thus, its long term use in equine practice could be of concern.     

Cone beam computed tomography and intraoral radiography for diagnosis of dental abnormalities in dogs and cats

The development of veterinary dentistry has substantially improved the ability to diagnose canine and feline dental abnormalities. Consequently, examinations previously performed only on humans are now available for small animals, thus improving the diagnostic quality. This has increased the need for technical qualification of veterinary professionals and increased technological investments. This study evaluated the use of cone beam computed tomography and intraoral radiography as complementary exams for diagnosing dental abnormalities in dogs and cats. Cone beam computed tomography was provided faster image acquisition with high image quality, was associated with low ionizing radiation levels, enabled image editing, and reduced the exam duration. Our results showed that radiography was an effective method for dental radiographic examination with low cost and fast execution times, and can be performed during surgical procedures.     

Molecular and serological detection of Babesia spp. in neotropical and exotic carnivores in Brazilian zoos

Large and small piroplasms have been observed in the blood smears of various wild carnivores, but few studies utilizing molecular characterization have been done. The goal of this present study was to investigate the presence of Babesia sp. by molecular and serologic techniques in exotic and neotropical carnivores maintained in captivity at Brazilian zoos. Blood and sera samples were collected from 146 Brazilian wild felids, 21 exotic felids, 1 genet (Genetta tigrina), 3 European wolves (Canis lupus), and 94 Brazilian wild canids in Brazilian zoos in the S?o Paulo and Mato Grosso states and in the Federal District. A total of 53 wild felids (31.74%) and 10 wild canids (10.31%) were seropositive for Babesia canis by Indirect Fluorescent Antibody Test (IFAT). Antibodies were detected in ocelots, little-spotted cats, margays, pampas cats, jaguars, pumas, jaguarundis, crab-eating foxes, and bush dogs. Babesia sp. DNA, with high similarity to B. leo, was detected in one pampas cat and one genet.     

Ultrasonographic assessment of maternal cardiac function and peripheral circulation during normal gestation in dogs

The aim of this study was to describe changes in cardiac morphology, systolic function and some peripheral hemodynamic parameters during normal pregnancy in dogs. Twenty healthy bitches, 10 pregnant (PG) and 10 non-pregnant controls (CG), were evaluated every 10 days using echocardiography from day 0 of the estrus cycle to parturition or to day 65 for the PG and CG groups, respectively. Systolic blood pressure (SBP) and uterine artery resistance index (RI) were also assessed. Throughout the study, the shortening fraction and cardiac output increased up to 30% vs. 5% (P<0.01) and 45% vs. 2% (P<0.01) in the PG and CG groups, respectively. In contrast, SBP and RI diminished up to 20% vs. 1% (P<0.01) and 29% vs. 0% (P<0.01) in the PG and CG groups, respectively. In conclusion, a decrease in afterload, an increase in cardiac output and cardiac hypertrophy appear to be the result of the hemodynamic modifications occurring during pregnancy in dogs.     

Studies on the development of a mating disruption system to control the tomato leafminer, Tuta absoluta Povolny (Lepidoptera: Gelechiidae)

BACKGROUND: The tomato leafminer (Tuta absoluta Povolny) has rapidly colonised the whole Mediterranean and South‐Atlantic coasts of Spain, and it has become a key problem in both outdoor and greenhouse crops. New control methods compatible with biological control are required, and mating disruption appears to be a perfect method in current agriculture, as it is an environmentally friendly and residue‐free technique. IPM packages tested have included the use of pheromones to detect populations, but there has not been much previous research on mating disruption of T. absoluta. In this work, pheromone doses varying from 10 to 40 g ha^?1, emitted at a constant rate over 4 months, were tested in greenhouses with different levels of containment in order to evaluate the efficacy of mating disruption on T. absoluta. RESULTS: Trials on containment level revealed that the flight of T. absoluta was satisfactorily disrupted with an initial pheromone dose of 30 g ha^?1, and levels of damage did not significantly differ from those in reference plots with insecticide treatments. Later efficacy trials confirmed previous experiences, and release studies showed that control of damage and flight disruption were taking place when releasing at least 85 mg pheromone per ha per day. CONCLUSION: Effective control using pheromone application against T. absoluta can be achieved, in greenhouses with high containment levels, for 4 months, with initial doses of 30 g ha^?1. Further research must be conducted in order to evaluate the prospect of outdoor application of mating disruption systems. Copyright © 2011 Society of Chemical Industry     

Health-promoting compounds in broccoli as influenced by refrigerated transport and retail sale period

Total aliphatic and indole glucosinolates, phenolic compounds (flavonoids and hydroxycinnamoyl derivatives), and vitamin C contents were evaluated in freshly harvested broccoli (Brassica oleracea L., var. italica, cv. Marathon) inflorescences. These were film-wrapped and stored for 7 days at 1 degrees C to simulate a maximum period of commercial transport and distribution. After cold storage, inflorescences were kept for 3 days at 15 degrees C to simulate a retail sale period. For wrapping, low-density polyethylene (LDPE) of 11 microm thickness was used. Gas composition was about 17% O(2) and 2% CO(2) during cold storage and about 16% O(2) and 3-4% CO(2) during shelf life within packages. The predominant glucosinolates were 4-methylsulfinylbutyl-glucosinolate (glucoraphanin), 3-indolylmethyl-glucosinolate (glucobrassicin), and 1-methoxy-3-indolylmethyl-glucosinolate (neoglucobrassicin). The predominant hydroxycinnamoyl derivatives were identified as 1,2,2'-trisinapoylgentiobiose, 1,2-diferuloylgentiobiose, 1,2'-disinapoyl-2-feruloylgentiobiose, and 3-O-caffeoyl-quinic (neochlorogenic acid). Results showed major losses at the end of both periods, in comparison with broccoli at harvest. Thus, the respective losses, at the end of cold storage and retail periods, were 71-80% of total glucosinolates, 62-59% of total flavonoids, 51-44% of sinapic acid derivatives, and 73-74% caffeoyl-quinic acid derivatives. Slight differences in all compound concentrations between storage and retail sale periods were detected. Distribution and retail periods had minimal effects on vitamin C. Weight loss was monitored at the end of both periods.     

Antiradical activity of water soluble components in common diet vegetables

The antiradical activity of water-soluble components contained in mushrooms (Psalliota campestris), onions (Allium cepa), white cabbage (Brassica oleracea var. alba), and yellow bell peppers (Capsicum annuum) against hydroxyl radicals was tested in a chemical and biological system. The vegetable juices were obtained by centrifugation of a vegetable homogenate processed at 2 degrees C or heated at 102 degrees C. The chemical system consisted of a buffered reaction mixture composed of Fe(III)-EDTA, 2-deoxy-D-ribose, ascorbic acid, and H(2)O(2) generating the hydroxyl radical. The antiradical activity was expressed as an inhibition of deoxyribose degradation. The biological system consisted of IMR32 neuroblastoma cells exposed to H(2)O(2) in the presence or absence of the vegetable juices. Cells were pretreated for either 24 h or 1 h with the vegetable juices, and reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used as a cell viability assay. All vegetable juices inhibited the degradation of deoxyribose and increased the viability of H(2)O(2) treated cells. Raw mushroom juice proved to be the most active in both cases. Boiling significantly affected the activity of mushroom juice, but did not change significantly the effect on onions and yellow bell peppers, and partially increased the activity of white cabbage juice. Mushroom antiradical activity was also confirmed by a cytofluorimetric analysis.     

Improved method to obtain pure alpha-galactosides from lupin seeds

Improvement of a previously described method of purification of alpha-galactosides, members of the raffinose family of oligosaccharides (RFOs), has been developed for lupins. The considerable amount of sucrose present in the RFO preparations obtained by the previous method has been removed by modifying the purification stage on diatomaceous earth and charcoal. The present method allows for the preparation of high-purity RFOs containing approximately 99.4% of RFOs in the form of a white fine powder, which provides new perspectives for the production of pure alpha-galactoside preparations for their use as prebiotics in functional foods.     

Rapid liquid chromatography tandem mass spectrometry assay to quantify plasma (-)-epicatechin metabolites after ingestion of a standard portion of cocoa beverage in humans

A rapid liquid chromatography electrospray ionization tandem mass spectrometry with negative ion detection method was developed and validated to determine cocoa flavonoid metabolites in human plasma and urine after the intake of a standard portion of a cocoa beverage. A chromatographic run time of only 9 min provided clear separation of all metabolites and internal standards. Samples were analyzed in a product-ion scan of m/z 289, 369, and 465 to identify the metabolites and in multiple reaction monitoring acquisition mode to quantify (-)-epicatechin ((-)-Ec) (289/ 245), (-)-epicatechin-glucuronide ((-)-EcG) (465/289), and (-)-epicatechin-sulfate ((-)-EcS) (369/289). One (-)-Ec-G and three (-)-Ec-S were identified and confirmed in urine as the major metabolites, and one (-)-Ec-G was the only metabolite present in plasma volunteers (n = 5) at a mean concentration of 625.7 +/- 198.3 nmol/L at 2 h after consumption of a cocoa beverage containing 54.4 mg of (-)-Ec.