C-reactive protein measurement in canine saliva.

An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.     

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Genetic basis and origin of resistance to acetolactate synthase inhibitors in Amaranthus palmeri from Spain and Italy

BACKGROUND

Amaranthus palmeri is an aggressive annual weed native to the United States, which has become invasive in some European countries. Populations resistant to acetolactate synthase (ALS) inhibitors have been recorded in Spain and Italy, but the evolutionary origin of the resistance traits remains unknown. Bioassays were conducted to identify cross-resistance to ALS inhibitors and a haplotype-based genetic approach was used to elucidate the origin and distribution of resistance in both countries.

RESULTS

Amaranthus palmeri populations were resistant to thifensulfuron-methyl and imazamox, and the 574-Leu mutant ALS allele was found to be the main cause of resistance among them. In two Spanish populations, 376-Glu and 197-Thr mutant ALS alleles were also found. The haplotype analyses revealed the presence of two and four distinct 574-Leu mutant haplotypes in the Italian and Spanish populations, respectively. None was common to both countries, but some mutant haplotypes were shared between geographically close populations or between populations more than 100 km apart. Wide genetic diversity was found in two very close Spanish populations.

CONCLUSION

ALS-resistant A. palmeri populations were introduced to Italy and Spain from outside Europe. Populations from both countries have different evolutionary histories and originate from independent introduction events. ALS resistance then spread over short and long distances by seed dispersal. The higher number and genetic diversity among mutant haplotypes from the Spanish populations indicated recurrent invasions. The implementation of control tactics to limit seed dispersal and the establishment of A. palmeri is recommended in both countries. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Irrigation scheduling of plastic greenhouse vegetable crops based on historical weather data

Irrigation scheduling based on the daily historical crop evapotranspiration (ET_h) data was theoretically and experimentally assessed for the major soil-grown greenhouse horticultural crops on the Almería coast in order to improve irrigation efficiency. Overall, the simulated seasonal ET_h values for different crop cycles from 41 greenhouses were not significantly different from the corresponding values of real-time crop evapotranspiration (ET_c). Additionally, for the main greenhouse crops on the Almería coast, the simulated values of the maximum cumulative soil water deficit in each of the 15 consecutive growth cycles (1988–2002) were determined using simple soil-water balances comparing daily ET_h and ET_c values to schedule irrigation. In most cases, no soil-water deficits affecting greenhouse crop productivity were detected, but the few cases found led us to also assess experimentally the use of ET_h for irrigation scheduling of greenhouse horticultural crops. The response of five greenhouse crops to water applications scheduled with daily estimates of ET_h and ET_c was evaluated in a typical enarenado soil. In tomato, fruit yield did not differ statistically between irrigation treatments, but the spring green bean irrigated using the ET_h data presented lower yield than that irrigated using the ET_c data. In the remaining experiments, the irrigation-management method based on ET_h data was modified to consider the standard deviation of the inter-annual greenhouse reference ET. No differences between irrigation treatments were found for productivity of pepper, zucchini and melon crops.     

Astaxanthin from Haematococcus pluvialis Prevents Oxidative Stress on Human Endothelial Cells without Toxicity

Astaxanthin, a powerful antioxidant, is a good candidate for the prevention of intracellular oxidative stress. The aim of the study was to compare the antioxidant activity of astaxanthin present in two natural extracts from Haematococcus pluvialis, a microalgae strain, with that of synthetic astaxanthin. Natural extracts were obtained either by solvent or supercritical extraction methods. UV, HPLC-DAD and (HPLC-(atmospheric pressure chemical ionization (APCI)+)/ion trap-MS) characterizations of both natural extracts showed similar compositions of carotenoids, but different percentages in free astaxanthin and its ester derivatives. The Trolox equivalent antioxidant capacity (TEAC) assay showed that natural extracts containing esters displayed stronger antioxidant activities than free astaxanthin. Their antioxidant capacities to inhibit intracellular oxidative stress were then evaluated on HUVEC cells. The intracellular antioxidant activity in natural extracts was approximately 90-times higher than synthetic astaxanthin (5 µM). No modification, neither in the morphology nor in the viability, of vascular human cells was observed by in vitro biocompatibility study up to 10 µM astaxanthin concentrations. Therefore, these results revealed the therapeutic potential of the natural extracts in vascular human cell protection against oxidative stress without toxicity, which could be exploited in prevention and/or treatment of cardiovascular diseases.     

Antioxidant activity of coffee model systems

Coffee model systems prepared from combinations of chlorogenic acid (CGA), N(alpha)-acetyl-1-arginine (A), sucrose (S), and cellulose (C) were roasted at 240 degrees C for 4 min prior to analysis by UV-visible spectrophotometry, capillary zone electrophoresis (CZE), and the ABTS radical cation decolorization assay. The A/CGA/S/C and A/S/C systems were also fractionated by gel filtration chromatography. Antioxidant activity of the systems showed a positive, nonlinear relationship with the amount of CGA remaining after roasting. Sucrose degradation was a major source of color in the heated systems. There was no relationship between antioxidant activity and color generation.     

Evaluation of metabolic detoxifying enzyme activities and insecticide resistance in Frankliniella occidentalis

The western flower thrips Frankliniella occidentalis (Pergande) is a very significant pest of a number of different agricultural crops in the south-east of Spain. The importance of thrips as a pest is not due mainly to the direct damage inflicted on the plant, but to the loss in commercial value which occurs as a consequence of the development of dark spots caused by the tomato spotted wilt virus (TSWV) which they transmit. The economic threshold is therefore almost zero, which enhances the problems of resistance management. The present work is part of a global project that attempts to evaluate the status of insecticide resistance in field populations of thrips obtained from several agricultural crops. We have studied, in either individual or pooled insects, some enzyme systems classically related to detoxification of insecticides: esterase and glutathione-S-transferase (GST). The activity of these enzymes from laboratory populations selected with various classes of insecticides has also been measured using several appropriate substrates. An increase in GST mean activity was found in two field-collected strains. Differences in frequency distributions of esterase and GST activities were found for both field-collected strains and for a laboratory strain selected with acrinathrin. These activities were compared with those of a wild-type reference strain.     

Cell wall disassembly during papaya softening: Role of ethylene in changes in composition,pectin-derived oligomers (PDOs) production and wall hydrolases

Cell wall disassembly in ripening climacteric fruit is a highly complex process where ethylene plays a crucial role. Ethylene inhibitors can be used to explore the changes in the cell wall matrix and cross-linked polysaccharides in ethylene-regulated processes. The results of applying the ethylene receptor blocking inhibitor 1-methylcyclopropene (1-MCP) and the ethylene-releasing compound ethephon (2-chloroethylphosphonic acid) indicate that softening of ‘Maradol’ papaya fruit is dependent on ethylene. When fruit were induced to ripen extensively by exposure to a high dose of ethephon, 1-MCP inhibited the subsequent softening dramatically, but when inhibition of the ethylene response was caused by application of 1-MCP, subsequent fruit treatment with ethephon promoted extensive loss of galactose from the water-soluble polysaccharides, but this was not accompanied by fruit softening. The cell wall changes accompanying normal fruit softening were pectin solubilization and polyuronide depolymerization and these processes occurred simultaneously. Polygalacturonase likely is responsible for the ripening-associated changes in ‘Maradol’ papaya fruit texture and pectin polymer integrity. An increase in extractable fruit polygalacturonase follows the increased presence of pectin-derived oligosaccharides.     

Afforestation trials with Prosopis chilensis (Mol.) Stuntz and Prosopis flexuosa D.C. in the Dry Chaco, Argentina

In implantation trials carried out with Prosopis chilensis and Prosopis flexuosa, the following variables were considered: implantation time (December and March), implantation technique (direct seeding and seedling transplantation) and age of the seedlings at the time of transplantation (60, 105 and 150 days). Survival was evaluated for more than 9 months after implantation and trials were repeated in 3 consecutive years.

Results from both species were similar and they show that survival after transplantation was higher (60–80%) than from direct seeding (35–45%). Survival from transplantation was independent of the size and age of the seedlings and it was less affected by climatic conditions than direct field sowing. Survival from direct seeding was higher in March than in December. The best time for either implantation alternative would be the end of the summer or beginning of fall, when climatic conditions increase the probability of seedling survival.