Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.     

MODELING GROWTH AND ION CONCENTRATION OF LILIUM IN RESPONSE TO NITROGEN:POTASSIUM:CALCIUM MIXTURE SOLUTIONS

Nutrient solution composition plays an important role in root uptake rate due to interactions among nutrients and internal regulation. Studies to determine the optimum nutrient solution concentration are focused on individual ions, ignoring the adaptation mechanisms triggered by plants when growing in a varying external nutrient concentration. The objective of the present study was to determine the response in growth and tissue ion concentration of lilium cv. ‘Navona’ to nutrient mixtures of varying proportions of nitrogen (N), potassium (K⁺), and calcium (Ca²⁺) in solution using mixture experiments methodology in order to determine the optimum concentration. Bulbs of lilium were transplanted in plastic crates and drip-irrigated with the treatment solutions, which consisted of a mixture of N, K⁺, and Ca²⁺ whose total concentration was 340 mg L^?1 and minimum concentrations of each ion was 34 mg L^?1. Chlorophyll concentration (SPAD), shoot fresh weight (FW), leaf FW, and leaf area were measured 60 days after transplanting and ion analysis was performed on shoot tissues from selected treatments. Lilium exhibited a moderate demand for N and K⁺ (136–170 mg L^?1 N and 116–136 mg L^?1 K⁺) and a very low demand for Ca²⁺ (34–88 mg · L^?1). This low demand may be due to the remobilization of the nutrients stored in the bulbs. Integrating the predictions of the models estimated to produce >90% of maximum growth, the optimum nutrient solution should contain Ca²⁺ at a concentration between 34 and 126 mg · L^?1, K⁺ between 119 and 211 mg · L^?1, and N between 92 mg · L^?1 and 211 mg · L^?1. Increasing external N concentration affected internal N concentration but not internal K⁺ or Ca²⁺ concentrations, despite that the increase in external N was associated with a decrease in external K⁺ and Ca²⁺. Similar trends were observed for external K⁺ and Ca²⁺ concentration. In conclusion, lilium was able to maintain a relatively constant K⁺ and Ca²⁺ concentration regardless of the lower concentration in the nutrient solution when N was increased (similar response was observed for K⁺ and Ca²⁺) and it has a low Ca²⁺ demand and moderate N and K⁺ supply.     

Weed abundance and soil seedbank responses to tillage systems in continuous maize crops

Weed abundance in crops undergoes frequent changes, often due to changes in tillage practices. Annual species, with quick germination, a short vegetative stage, profuse seed production and long-lived seeds become problematic under zero-tillage systems. Portulaca oleracea L. and Amaranthus blitoides L. are widespread weeds in the Mediterranean area, prominent in irrigated crops. We studied the total weed abundance in the field, and specifically these two species (Portulaca oleracea and Amaranthus blitoides) with high frequency of occurrence in monoculture maize, from 2012 to 2014, in the field and soil seedbank. Results showed significant differences between zero-tillage (ZT) and conventional tillage (CT) systems on total weed abundance and relative abundance of Portulaca oleracea. Total weed abundance decreased in ZT plots (from 136 plants m^?2 to 25 and 46 plants m^?2, in 2013 and 2014 respectively). The same trend was observed in Portulaca oleracea recorded in ZT plots, but the abundance of Amaranthus blitoides did not vary in this system. Weed seedling germination and weed seed numbers both of total weed seedbank and Portulaca oleracea, were greater in ZT plots compared to CT, regarding Amaranthus blitoides seedling germination and seed count, the values did not increase with ZT, in continuous maize crops.     

Atmospheric Mercury Fluxes in a Southern Boreal Forest and Wetland

Total gaseous mercury (TGM) fluxes from the forest floor and a boreal wetland were measured by a flux chamber technique coupled with an automatic mercury vapour analyser. The fluxes were measured at three sampling sites in southern Finland, 61°14′ N, 25°04′ E in summer 2007, with additionally in situ TGM concentrations in the air at one of the sites and mercury bulk deposition at another. Most of the flux data were collected during the daytime. At one of the sites, diurnal flux behaviour was studied, and a clear cycle with an afternoon maximum and a night minimum was observed. The highest emissions (up to 3.5 ng m⁻² h⁻¹) were observed at the forest floor site having a moss and grass cover. At the wetland and litter-rich forest floor sites, the emissions were below 1 ng m⁻² h⁻¹ and sometimes negative (down to −1.0 ng m⁻² h⁻¹), indicating mercury uptake. The measured average fluxes in August were 0.9 ± 1.1 and 0.2 ± 0.3 ng m⁻² h⁻¹ for the forest floor sites and wetland sites, respectively. The flux data were compared with the mercury bulk deposition, which proved to be of the same magnitude, but opposite in sign. At the mossy forest floor site, the extrapolated TGM emissions were 130% of the Hg deposition in August 2007. Comparison with other studies showed that the fluxes in background areas are relatively uniform, regardless of measurement site location and method used. Airborne TGM remained at the background level during the study, with an average value of 1.3 ± 0.2 ng m⁻³; it frequently showed a diurnal cycle pattern.     

Tomato fruit quality as influenced by the interactions between agricultural techniques and harvesting period

Tomato (Solanum lycopersicum Mill) is an important crop in terms of its economic and nutritional value. Many factors, including cultivar, climate, geography, geochemistry, and agricultural practice, can affect its nutrient concentrations. An HJ‐biplot study was performed to examine the effects of cultivar (Dorothy, Boludo, Dominique, Thomas, and Dunkan), agricultural practices, climatic factors, and their interactions. Significant differences were analyzed using a one‐way ANOVA. All samples were collected and assayed at the same degree of ripeness. In the conventional and organic tomato samples, those harvested from December to April had the highest concentrations of fructose, glucose, citric acid, malic acid, ascorbic acid, protein, Na, and Mg, while those harvested in October had the highest concentrations of lycopene and hydroxycinnamic acid. There were high concentrations of Ca, P, Zn, and Cu in the no‐soil tomatoes. Conventional and organic cultivation practices showed similar results with respect to the collection period, both presenting high organic compound concentrations, while high mineral concentrations seemed to correspond to the no‐soil practice. No clear pattern was observed among the different cultivars, perhaps due to all the samples having been collected at the same degree of ripeness.     

Effect of Preharvest Sprays of Calcium on Cracking and ‘Schattenmorelle’ Sour Cherry Fruit Quality Harvested Mechanically

The aim of the study was to examine effect of preharvest sprays of calcium (Ca) in the form of Ca-chloride (CaCl₂), Ca-nitrate [Ca(NO₃)₂], or a mixture of Ca-formate, Ca-acetate, CaCl₂, and Ca(NO₃)₂ on cracking and quality of ‘Schattenmorelle’ sour cherry fruit harvested mechanically. The experiment was conducted in 2008–2009 at a commercial orchard in central Poland. Mature trees grew on a coarse-textured soil poor in organic matter, at a spacing of 4.0 × 1.5 m. The spray treatments of Ca were performed at 7-day intervals, starting 28 days before harvest, at the rates of 5.0–5.6 kg Ca ha^?1 per season. The trees sprayed with water were treated as the control. Fruit were harvested mechanically when peduncle-fruit detachment force dropped below 3 N. The results showed that preharvest Ca sprays caused no leaf damage. This measure did not affect yield, mean fruit weight, soluble solids concentration and titratable acidity of fruit, and weight loss of fruit during 24 h after harvest. Fruit sprayed with Ca had improved status of this nutrient, and were less liable to juicy leakage from the stem scar, rain-induced cracking, and preharvest decay caused by Glomerella cingulata. The above effects of Ca sprays did not depend on the tested material. It was concluded that preharvest sprays of Ca as CaCl₂ and/or Ca(NO₃)₂ should be recommended in ‘Schattenmorelle’ sour cherry orchards to reduce fruit losses resulting from rain-induced cracking, leakage of juice, and the incidence of cherry bitter rot.     

Heterogeneous aspects of lipid oxidation in dried microencapsulated oils

This work was aimed at studying lipid oxidation in dried microencapsulated oils (DMOs) during long-term storage. Samples were prepared by freeze-drying of emulsions containing sodium caseinate and lactose as encapsulating components. Evaluation of lipid oxidation was approached by quantitative analysis of nonvolatile lipid oxidation products and tocopherol. Lipid oxidation products were analyzed by separation of polar compounds by adsorption chromatography followed by HPSEC with refraction index detection for quantitation of oxidized triglyceride monomers, dimers, and oligomers. The analytical method applied enabled the detection of different oxidative patterns between the free and encapsulated oil fractions. The free oil fraction of DMOs showed a typical oxidative pattern for oils in continuous phase, which consisted of a clear induction period, in which hydroperoxides (oxidized triglyceride monomers) accumulated, before oxidation accelerated. The end of the induction period was marked by the total loss of tocopherol and the initiation of polymerization. On the contrary, the encapsulated oil showed a pattern characteristic of a mixture of oils with different oxidation status. Thus, high contents of advanced oxidation compounds (polymerization compounds) were detected when the antioxidant (tocopherol) was still present in high amounts. It is concluded that the encapsulated oil was comprised of oil globules with very different oxidation status. The results obtained in this study gave evidence of heterogeneous aspects of lipid oxidation in a dispersed-lipid food system.     

Food consumption patterns in an adult urban population in Beirut, Lebanon

OBJECTIVES: To investigate, measure and assess the food consumption pattern of the adult population living in Beirut, Lebanon, and to identify inadequate or excessive intake of food groups particularly linked to non-communicable diseases. DESIGN: A cross-sectional food consumption survey was conducted in 2001. Dietary habits were assessed by means of a quantitative food-frequency questionnaire. SETTING: Dietary survey of the urban population (Beirut). SUBJECTS: Random sample of 444 adult subjects (aged 25-54 years) in Beirut. RESULTS: The mean consumption of food by the study population was estimated to be 3,030 g day(-1), providing an energy intake of 2,523.57 kcal day(-1). Fat contributed 38.9% to the average daily energy intake, protein 13.4% and carbohydrates 47.2%. Mean consumption of fruits and vegetables was approximately 367 g day(-1) and 45.3% of subjects consumed less than the recommended 400 g daily. Cereals contributed 324.5 g day(-1), providing 35.0% of daily energy intake, with bread being the most highly consumed (146.2 g day(-1)) in this food group. The mean intake of meat and poultry products was 91.7 g day(-1) and provided 8.8% of daily energy intake, with consumption of butchery products especially beef being the highest (47.6 g day(-1)) followed by poultry (36.1 g day(-1)). A low consumption of fish was noted (19.7 g day(-1)), with 73.6% of subjects consuming less than the recommended 2 servings of fish per week. Dairy products contributed 243.1 g day(-1) or 10.9% of daily energy intake, and milk was the least consumed dairy product (56.8% of consumers). The intake of added fats and oils, excluding those in cooked recipes, was 20.4 g day(-1); olive oil was not used in cooking but was added solely at the table and its mean intake was 5 g day(-1). The consumption of butter was low (0.86 g day(-1)) and vegetable oil was the type of fat mostly used in cooking. The average intake of alcoholic beverages was low (33.6 g day(-1)), accounting for 0.7% of total energy intake. Women had significantly higher intakes of milk, dairy products, vegetables and coffee than men (P < 0.05). The percentage of women who reported the use of low-fat items was significantly higher than that of men. Younger people (25-34 years) ate significantly more meat, sugar, alcoholic beverages and soft drinks, and consumed significantly less cooked vegetables and legumes, than older ones (P < 0.05). CONCLUSIONS: The rather high contribution of fat to daily energy intake, the low intake of fish and the relatively high percentage of people consuming less than the recommended amount of fruits and vegetables observed in this study suggest that the adult Lebanese population is at increased risk of cardiovascular diseases, obesity and other non-communicable diseases, which provides the basis for recommending increased intakes of fish, particularly fatty fish, and fruits and vegetables.     

Use of Aloe vera gel coating preserves the functional properties of table grapes

Table grapes (Vitis vinifera L. cv. Crimson Seedless) were coated with Aloe vera gel according to our developed patent (SP Patent P200302937) and then stored for 35 days at 1 degrees C, and the subsequent shelf life (SL) was monitored at 20 degrees C. Uncoated clusters showed a rapid loss of functional compounds, such as total phenolics and ascorbic acid. These changes were accompanied by reduction of the total antioxidant activity (TAA) and increases in total anthocyanins, showing an accelerated ripening process. On the contrary, table grapes coated with Aloe vera gel significantly delayed the above changes, such as the retention of ascorbic acid during cold storage or SL. Consequently, Aloe vera gel coating, a simple and noncontaminating treatment, maintained the functional properties during postharvest storage of table grapes.     

Effects of rhodium heterogeneous catalyst and isomerization conditions on linoleic acid conjugation of soybean oil

Rhodium heterogeneous catalyst was used to catalyze isomerization of linoleic acid in soybean oil to conjugated linoleic acid (CLA). A central composite rotatable design with five levels of three variables, namely, reaction temperature, stirring speed, and reaction time, was used to determine the maximum CLA yield. The formation of CLA during isomerization was greatly dependent on the reaction temperature and time. The CLA content of soybean oil increased from 0.63 to 202.42 mg/g oil when isomerization was done at 200 degrees C, with a stirring speed of 200 rpm for 49 min. Analysis of triacylglycerol positions showed that linoleic acid at any position in a triacylglyceride could possibly be isomerized to CLA.