An enrichment microsphere immunoassay for the detection of <Emphasis Type="Italic">Pectobacterium atrosepticum</Emphasis> and <Emphasis Type="Italic">Dickeya dianthicola</Emphasis> in potato tuber extracts

An enrichment microsphere immunoassay (MIA) was developed, based on the Luminex xMAP® technology, for the simultaneous (duplex) detection of Pectobacterium atrosepticum (former name Erwinia carotovora subsp. atroseptica) (Pca) and Dickeya dianthicola (former name Erwinia chrysanthemi) (Dcd) in potato plant extracts. Target bacteria in the extracts were enriched for 48 h in a semi-selective broth containing polypectate under low oxygen conditions. Samples were subsequently incubated with antibody-coated colour-coded microspheres (beads) and with secondary antibodies conjugated with Alexa Fluor® 532, a reporter dye. Samples were analyzed with the Luminex analyzer, in which one laser identified each microsphere and another laser the reporter dye conjugated to the secondary antibodies. The assay required minimal sample preparation, could be completed in 1 h, was performed in 96 wells microtitreplates and required no wash steps. The limit of detection for the duplex enrichment MIA was 100–1000 cfu ml^?1, which was a hundred times lower than of an enrichment-ELISA. Without enrichment, the sensitivity of MIA and ELISA was largely similar and ranged between 10⁶ and 10⁷ cells ml^?1. No difference in sensitivity was found between a MIA in a single or duplex format. In a comparative test with non-infected potato plant extracts and extracts from plants infected with Pca or Dcd, results of the enrichment MIA correlated well with those of the enrichment ELISA and enrichment PCR. These results indicate that MIA can be reliably used for multiplex detection of soft rot Enterbacteriaceae in crude potato plant extracts. The technology is an attractive and cost-effective alternative to other detection methods, including ELISA.     

Gross nitrogen mineralisation and fungi-to-bacteria ratios are negatively correlated in boreal forests

In terrestrial ecosystems, gross nitrogen mineralisation is positively correlated to microbial biomass but negatively to soil organic matter C-to-N ratios; the influence of the microbial community structure is less well known. Here, we relate rates of gross N mineralisation to fungi-to-bacteria ratios in three natural forest types of contrasting N availability and in a long-term N-loading experiment in a boreal forest. We report, for the first time, a strong negative correlation between gross N mineralisation and the fungi-to-bacteria ratio ( = 0.91, P = 0.0005, N = 7). There was also a negative correlation between gross N mineralisation and the C-to-N ratio ( = 0.89, P = 0.001, N = 7), but a weaker positive correlation between gross N mineralisation and soil pH ( = 0.64, P = 0.019, N = 7). Our analysis suggests that soil fungi-to-bacteria and C-to-N ratios are interrelated and that they exert strong influences on soil N cycling in boreal forests.     

Autoimmune hepatitis-specific antibodies against soluble liver antigen and liver cytosol type 1 in patients with chronic viral hepatitis

Background

Non-organ specific autoantibodies are highly prevalent in patients with chronic hepatitis C (HCV). Among them, anti-liver kidney microsomal type 1 (LKM1) antibody – the serological marker of type 2 autoimmune hepatitis (AIH-2)- is detected in up to 11% of the HCV-infected subjects. On the other hand, anti-liver cytosol type 1 antibodies (anti-LC1) – either in association with anti-LKM1, or in isolation- and anti-soluble liver antigen antibodies (anti-SLA) have been considered as useful and specific diagnostic markers for AIH. However, their specificity for AIH has been questioned by some recent studies, which have shown the detection of anti-LC1 and anti-SLA by immunoprecipitation assays in HCV patients irrespective of their anti-LKM1 status. The aim of the present study was to test the anti-LC1 and anti-SLA presence by specific enzyme linked immunosorbent assays (ELISAs), in a large group of Greek HCV-infected patients with or without anti-LKM1 reactivity as firstly, immunoprecipitation assays are limited to few specialized laboratories worldwide and cannot be used routinely and secondly, to assess whether application of such tests has any relevance in the context of patients with viral hepatitis since antibody detection based on such ELISAs has not been described in detail in large groups of HCV patients.

Methods

One hundred and thirty eight consecutive HCV patients (120 anti-LKM1 negative and 18 anti-LKM1 positive) were investigated for the presence of anti-LC1 and anti-SLA by commercial ELISAs. A similar number (120) of chronic hepatitis B virus (HBV) infected patients seronegative for anti-LKM1 was also tested as pathological controls.

Results

Six out of 18 (33%) anti-LKM^pos/HCV^pos patients tested positive for anti-LC1 compared to 1/120 (0.83%) anti-LKM^neg/HCV^pos patients and 0/120 (0%) of the anti-LKM1^neg/HBV^pos patients (p < 0.001 for both comparisons). Anti-SLA antibodies were not present in any of the HCV (with or without anti-LKM1) or HBV-infected patients.

Conclusion

We showed that anti-LC1 and anti-SLA autoantibodies are not detected by conventional assays in a large group of anti-LKM1 negative patients with chronic hepatitis B and C infections. Based on these results we cannot find any justification for the application of anti-LC1 and anti-SLA tests in the routine laboratory testing of viral hepatitis-related autoantibody serology with the only potential exception being the anti-LC1 screening in anti-LKM1^pos/HCV^pos patients.

     

Effect of fibers on the flow property of turbulent fiber suspensions in a contraction

In the flow of turbulent fiber suspensions flowing through a contraction with rectangular cross-section, the Reynolds averaged Navier-Stokes equation with the term of additional stress resulting from fibers was solved with the Reynolds stress model to get distributions of the mean velocity, mean pressure, turbulent kinetic energy and turbulent dissipation. It is found that the mean velocities at exit are small around the center and large near the wall for higher concentration. Fibers reduce turbulent intensity and turbulent dissipation at central line, but enhance them over the cross section at exit. Fibers have no effect of restraint on the turbulence in the contraction flow. The additional stress resulting from fibers plays a role in the increase of drag.     

Chemical Composition of Green Tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis>) Infusions Commercialized in Portugal

To evaluate the potential benefits and risks associated with tea consumption it is important to identify the constituents of this beverage. Levels of some minerals, caffeine and catechins in green tea samples commercialized in Portugal were evaluated. Potassium is the metal present in larger amount (92–151 mg/l). The content of sodium, calcium, fluoride, aluminium, manganese and iron were 35–69, 1.9–3.5, 0.80–2.0, 1.0–2.2, 0.52–1.9, 0.020–0.128 mg/l, respectively. Chromium and selenium were not detected. The resulting data showed considerable variability in catechins content. The levels of epigallocatechin gallate (EGCG) ranged from 117 to 442 mg/l, epicatechin 3-gallate (EGC) from 203 to 471 mg/l, epigallocatechin (ECG) from 16.9 to 150 mg/l, epicatechin (EC) from 25 to 81 mg/l and catechin (C) from 9.03 to 115 mg/l. Caffeine contents in the green tea infusions studied were between 141–338 mg/l. Green tea infusions provide significant amounts of catechins and could be an important source of some minerals.     

Antihyperglycemic Effects of <Emphasis Type="Italic">Platycodon grandiflorum</Emphasis> (Jacq.) A. DC. Extract on Streptozotocin-induced Diabetic Mice

The root of Platycodon grandiflorum (Jacq.) A. DC has been reported to have a wide range of health benefits in oriental food. This study examined the hypoglycemic effects of Platycodon grandiflorum (Jacq.) A. DC aqueous-ethanol extract (PGE) in streptozotocin (STZ) -induced diabetic ICR mice (STZ diabetic mice) for the first time. The effects of PGE on blood glucose, plasma insulin levels and body weight were investigated. A significant decrease in blood glucose levels was observed after single administration of PGE. Furthermore, Glibenclamide and PGE significantly suppressed the rise in blood glucose after 30 min in the acute glucose tolerance test. Treatment with glibenclamide and PGE resulted in a reduction in blood glucose levels from the 2nd week, and this reduction was maintained until the 4th week of treatment. The body weight changed slightly in glibenclamide and PGE treated mice in comparison with the STZ control group. Plasma insulin levels were increased with glibenclamide treatment in STZ diabetic mice, whereas such effect was not observed with PGE. These results indicated that PGE could induce hypoglycemic effects without stimulating insulin secretion.     

The Effect of Consumption of Selenium Enriched Rye/Wheat Sourdough Bread on the Body’s Selenium Status

The potential of selenium-enriched rye/wheat sourdough bread as a route for supplementing dietary selenium intakes is reported. In addition to their normal diets, 24 female volunteers (24 to 25 years old) were fed either selenium-enriched bread or non-enriched bread each day (68.02 and 0.84 μg selenium day⁻¹ respectively) for 4 weeks. The chemical form of the selenium in the bread had been characterised using HPLC-ICP-MS, which showed that 42% of the extractable selenium was present as selenomethionine. Plasma selenium levels and plasma platelet glutathione peroxidase (GPx1) activity were measured in the volunteers’ blood over a 6-week period. A statistically significant difference (p = 0.001) was observed in the mean percentage change data, calculated from the plasma selenium level measurements for the enriched and control group, over the duration of the study. A comparable difference was not observed for the platelet GPx1 activity (p = 0.756), over the same period. Two weeks after cessation of the feeding stage, i.e., at t = 6 weeks, the mean percentage change value for the selenium plasma levels in the enriched group was still significantly elevated, suggesting that the absorbed selenium had been incorporated into the body’s selenium reserves, and was then being slowly released back into the volunteers’ blood.     

Content of Nutrient and Antinutrient in Edible Flowers of Wild Plants in Mexico

Nutrient and antinutritional/toxic factors present in some edible flowers consumed in Mexico were determined. The edible flowers were: Agave salmiana, Aloe vera, Arbutus xalapensis, Cucurbita pepo (cultivated), Erythrina americana, Erythrina caribaea, Euphorbia radians benth and Yucca filifera. The nutrient content in the flowers studied is similar to that of the edible leaves and flowers studied mainly in Africa. The moisture content of the flowers varied from 860 to 932 g kg⁻¹. Crude protein (CP) was between 113 to 275 g kg⁻¹ DM, crude fiber, 104 to 177 g kg⁻¹ DM and the nitrogen free extract, between 425 to 667 g kg⁻¹ DM. The highest chemical score (CS) was found in E. americana and A. salmiana; in five samples the limiting amino acid was lysine, and in three of them it was tryptophan. Trypsin inhibitors and hemaglutinnins had a very low concentration. Alkaloids were present in both the Erythrina species and the saponins in A. salmiana and Y. filifera. Cyanogenic glucosides were not found in the studied flowers. The traditional process of preparing these specific flowers before consumption is by cooking them and discarding the broth; in this way the toxic substances are diminished or eliminated. These edible flowers from wild plants consumed in local areas of the country play an important role in the diet of the people at least during the short time of the season where they are blooming.     

Timing and order of different insecticide classes drive control of Drosophila suzukii; a modeling approach

The spotted-wing drosophila, Drosophila suzukii Matsumura, is an invasive pest causing significant damage to soft skinned fruits. Control of D. suzukii is critical since there is no tolerance for infested fruit in the market. While most insecticides control one or more D. suzukii life-stages (e.g., egg, larvae, and adult), the impact of insecticides that are toxic to immature stages  is unclear on the subsequent generation of a field population. Insecticides were applied at field recommended rates on cherries and blueberries in the laboratory to determine immature D. suzukii mortality. Spinetoram, cyantraniliprole, malathion, methomyl, spinosad, and phosmet resulted in relatively high mortality of all immature life stages. Zeta-cypermethrin, cyclaniliprole, and fenpropathrin resulted in lower mortality of egg and all larval instars. Malathion was also applied to lowbush blueberries with different fruit sizes (small, medium, and large) in the laboratory and there was no statistical difference in mortality rates depending on fruit sizes. Mortality data from the laboratory experiments were used to parameterize a refined D. suzukii population model. The model revealed that the timing and order of different insecticide classes are important to control D. suzukii population. Model runs that included early applications of more effective insecticides resulted in high immature mortality and greater reduction of D. suzukii populations compared to treatments applied later.

     

Identifying crop yield gaps with site- and season-specific data-driven models of yield potential

Precision Agriculture - There is considerable interest and value in identifying the gap between crop yields that have actually been achieved, and yields that could have potentially been achieved. A...