Use of electrochemical biosensor and gas chromatography for determination of dichlorvos in wheat

Two methods for the determination of dichlorvos in durum wheat by electrochemical assay and gas chromatography, respectively, have been developed. Dichlorvos, an organophosphorus anticholinesterase pesticide, was extracted from wheat with hexane, and the filtered extract was directly analyzed by gas chromatography with nitrogen-phosphorus flame detection (NPD). Recoveries of dichlorvos from milled wheat spiked at 0.25-1.5 microg/g ranged from 96.5 to 100.9%, and the limit of detection was 0.02 microg/g. The electrochemical assay was based on the detection of choline, the acetylcholinesterase product, via a choline oxidase biosensor. An aliquot of the filtered hexane extract was partitioned with phosphate buffer solution, and the organic layer was evaporated prior to electrochemical analysis. A limit of detection of 0.05 microg/g of dichlorvos was obtained with mean recoveries of 97-103% at spiking levels of 0.25-1.5 microg/g. A good correlation (r = 0.9919) was found between the results obtained with the electrochemical and those obtained with the gas chromatographic methods. The electrochemical method was peer-validated in two laboratories that analyzed 10 blind samples (5 duplicates), including a blank and 4 spiked samples with dichlorvos at levels of 0.25, 0.60, 1.00, and 1.50 microg/g. Within-laboratory repeatability (RSDr) and between-laboratory reproducibility (RSDR) ranged from 5.5 to 7.8% and from 9.9 to 17.6%, respectively.     

Dietary Supplementation of Oregano and Sage Dried Leaves on Performances and Meat Quality of Rabbits

The aim of this research was to evaluate the dietary supplementation 1% （w/w） of oregano and sage dried leaves on performances and meat quality of broiler rabbits. A feeding trial, which lasted 48 d, was carried out on 105 male Bianca Italiana rabbits randomly divided in seven groups and fed ad libitum. At the end of the trial ten animals per group were slaughtered and samples of dorsal muscle were taken in order to perform laboratory analysis. Mortality rate did not statistically differ between groups. Growth performances of animals fed diets supplemented with aromatic plants were higher （P〈0.05） than those of animals of control group, whereas carcass parameters were not affected by treatments excepting for the slaughter weight that showed the same trend as growth performances. Meat quality traits, oxidative lipid stability and fatty acid proifle were not inlfuenced by aromatic plant supplementation. In conclusion, oregano and sage in form of dried leaves can be used in rabbit without adverse effects on performance, carcass characteristics and meat quality traits.     

A new set of microsatellite markers for Fragaria species and their application in linkage analysis

Summary

This study reports the development of 68 new microsatellite markers. Of these, 45 were obtained, together with 20 others already published, from an AC-enriched genomic library of the wild strawberry Fragaria vesca. The 68 markers were tested for transportability to the cultivated strawberry F. ananassa ‘Miss’ and 83% gave positive amplifications. Twenty pairs of primers were selected and tested for their transportability to 16 Fragaria taxa and eight species of Rosaceae (peach, almond, apricot, European and Sino-Japanese plums, sweet and sour cherry, apple). The average proportion of primers amplifying loci in Fragaria was 69%, while the transportability to Rosaceae was very low and resulted in null amplification for 80% of the primer pairs. In addition, 23 microsatellite markers were developed from F. ananassa ‘expressed sequence tags’ databases. A total of 141 primer pairs from these and published primers, were tested for polymorphism in the two parents (91.333.2 and ‘Snovit’; both belonging to F. vesca) of a full sib population of 46 individuals. Fifty-eight percent of the primers were discarded because they were monomorphic, or were difficult to interpret, or their allelic conformation was not useful for mapping. The segregation of 73 primers was tested in the progeny and a partial map of the female parent was constructed, based on the segregation of 66 useful markers that were ordered into eight linkage groups of which four had from seven to 14 markers.     

Perspectives on the use of a seaweed extract to moderate the negative effects of alternate bearing in apple trees

Summary

Alternate bearing is a serious problem in many apple varieties. In fact, each year, apple trees have to be thinned in order to ensure a correct and constant balance between the reproductive and vegetative tendencies of the plant. In organic fruit growing, there is no alternative to the expensive and time-consuming process of hand-thinning. Under these conditions, the use of natural bio-stimulants which enhance the efficiency of nutrient uptake, plant fitness to different pedoclimatic conditions, and plant tolerance to stress, offers new opportunities to reduce the negative effects of alternate bearing. The aim of this work was to evaluate the effects of Actiwave^®, a metabolic enhancer derived from the alga, Ascophillum nodosum, on ‘Fuji’ apple trees affected by experimentally-imposed alternate bearing. The experiments were carried in open field conditions over four consecutive years. Although the effect of Actiwave^® was erratic from year-to-year, the compound generally reduced those problems linked to alternate bearing. Actiwave^® had its most significant effect on alternate bearing plants that were also affected by nutrient deprivation due to the absence of fertilisation. In these trees,Actiwave^® decreased the oscillations in yield between “on” and “off” years and increased the average fruit weight on plants affected by too-high a crop load. Treated trees also showed higher leaf chlorophyll contents (increased by 12%), with a consequent increase in the rates of photosynthesis and respiration. On the other hand, under standard conditions, in well-fertilised balanced plants, Actiwave^® did not have any significant effect. This evidence corroborates the hypothesis that Actiwave^® may be a useful tool to reduce alternate bearing in organic and low-input farming.     

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.     

Clavibacter michiganensis subsp. michiganesis: Tracking strains using their genetic differentiations by ISSR markers in Southern Turkey

There is no available data published related to the dissemination of bacterial canker caused by Clavibacter michiganensis subsp. michiganensis (Cmm) and its genetic diversity in Turkey. It is important to control new introduced inoculum sources by commercial seeds and seedlings. Pathogens were identified by morphological features and the identity was confirmed by PCR amplification using a specific primer PSA-4, PSA-R in addition to microbiological tests. ISSR markers showing high polymorphism ratios were selected and used to characterize Cmm strains. The collected strains were classified into different groups on the basis of ISSR-PCR fingerprints, which showed remarkable genetic specificity and diversity not previously identified in Cmm, suggesting that genetic differences are related to dissemination of the pathogen in the region. This is the first ever study carried out on the characterization of Cmm using ISSR. The selected ISSR primers to characterize Cmm can be used to determine genetic differences in further studies.     

Lipoxygenase distribution in coffee (Coffea arabica L.) berries

In this paper lipoxygenase (LOX) presence was investigated in coffee berries to determine its involvement in lipid degradative metabolism of plants grown in organic and conventional cultivations. An immunochemical analysis has evidenced a ca. 80 kDa protein, cross-reacting with an anti-LOX antibody, only in the pulp fraction of berries obtained from plants of both cultivations. LOX activity in this fraction could be monitored either as conjugated diene formation or reaction products (determined by HPLC) and was mainly associated with a heavy membrane fraction (HMF, enriched in tonoplast, endoplasmic reticulum, plasma membrane, and mitochondria) and a light membrane fraction (LMF, enriched in plasma membrane and endoplasmic reticulum, with low levels of tonoplast and mitochondria). The LOX activity of LMF from berries of both cultivations showed an optimum at pH 8.0. The HMF exhibited a different activity peak in samples from conventional (pH 8.0) and organic (pH 5.5) cultures, suggesting the presence of different isoenzymes. These findings were also confirmed by variation of the ratio of 9- and 13-hydroperoxides in organic (1:1) and conventional cultivations (1:10), indicating that the organic one was subjected to an oxidative stress in the coffee pulp fraction leading to the expression of an acidic LOX. Such de novo synthesized LOX activity could be responsible for the production of secondary metabolites, which may interfere with the organoleptic profile of coffee.     

Salinity of nutrient solution influences the shelf-life of fresh-cut lettuce grown in floating system

Quality, microbiological and enzymatic characteristics of fresh-cut lettuce (Lactuca sativa var. longifolia, ‘Duende’), grown in floating system with three electrical conductivities of nutrient solutions (2.8, 3.8 and 4.8 mS cm^?1), were investigated in order to evaluate the effect of salinity on product shelf-life during cold storage (9 d at 4 °C). Pre-harvest salinity of 3.8 and 4.8 mS cm^?1 improved the properties of fresh-cut lettuce, since CO₂ production was reduced with a subsequent control of the decay process. Fresh-cut processing caused an activation of polyphenol oxidase and peroxidase; in all cases the product obtained by salinity treatments was less subject to oxidase activity and browning phenomena during storage. Increased salinity reduced the number of mesophilic bacteria and of moulds and yeasts, assessed by plate counts on different culture media; in contrast, Enterobacteriaceae levels were unaffected by pre-harvest treatments. The research demonstrated that an increase in nutrient solution electrical conductivity, through the use of floating system, affects fresh-cut lettuce characteristics, improving shelf-life of the product.     

Weak effects of habitat type on susceptibility to invasive freshwater species: an Italian case study

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