An enzyme-linked immunosorbent assay for detection of avian influenza virus subtypes H5 and H7 antibodies

Background

Avian influenza virus (AIV) subtypes H5 and H7 attracts particular attention because of the risk of their potential pathogenicity in poultry. The haemagglutination inhibition (HI) test is widely used as subtype specific test for serological diagnostics despite the laborious nature of this method. However, enzyme-linked immunosorbent assays (ELISAs) are being explored as an alternative test method.H5 and H7 specific monoclonal antibodies were experimentally raised and used in the development of inhibition ELISAs for detection of serological response specifically directed against AIV subtypes H5 and H7. The ELISAs were evaluated with polyclonal chicken anti-AIV antibodies against AIV subtypes: H1N2, H5N2, H5N7, H7N1, H7N7, H9N9, H10N4 and H16N3.

Results

Both the H5 and H7 ELISA proved to have a high sensitivity and specificity and the ELISAs detected H5 and H7 antibodies earlier during experimental infection than the HI test did. The reproducibility of the ELISA’s performed at different times was high with Pearson correlation coefficients of 0.96-0.98.

Conclusions

The ELISAs are a potential alternative to the HI test for screening of large amounts of avian sera, although only experimental sera were tested in this study.     

Pressurized liquid extraction and HPLC quantification of folic Acid in fortified wheat flours

A pressurized liquid extraction (PLE) method using phosphate buffer as solvent was applied for folic acid (FA) extraction from fortified wheat flours and was compared to a standard solid-liquid extraction (SLE) method. Extracted FA was quantified by reverse phase high-performance liquid chromatography (RP-HPLC) hyphenated with a phenyl column and an absorption photometric detector (λ = 280 nm). Detection and quantification limits were 0.12 and 0.4 ng, respectively, corresponding to 0.06 and 0.2 μg g(-1) of analyzed wheat flour. Equivalent FA contents were found by both extraction methods, but a single PLE allowed a total recovery of FA content, whereas at least three successive SLEs were needed to achieve a total recovery of FA. The obtained results indicated that PLE is a rapid and efficient technique for FA extraction from fortified wheat flour.     

Phylogeny of five predominant pospiviroid species in Belgium

In Belgium pospiviroids are routinely detected in various hosts. The most frequently found pospiviroids are: Citrus exocortis viroid (CEVd), Chrysanthemum stunt viroid (CSVd), Potato spindle tuber viroid (PSTVd), Tomato apical stunt viroid (TASVd) and Tomato chlorotic dwarf viroid (TCDVd). Apart from the high incidence of pospiviroids in latently-infected ornamentals, viroids have also been found in plants where they cause disease: PSTVd and TCDVd in tomatoes and CSVd in chrysanthemum. In order to gain more epidemiological data on these infections, this study has conducted phylogenetic analyses of Belgian isolates for each of these five pospiviroid species. PSTVd and CEVd-isolates show a clustering depending on host plant identity. This was not observed for TCDVd and TASVd. A very high degree of sequence similarity was noticeable for CSVd-isolates from various hosts. During the past decade, PSTVd and CSVd-infected mother plants have been systematically eradicated in Belgium after positive detection results, also when found in symptomless plants, leading to a decreased trend of these quarantine pests in the past few years. However, other non-quarantine pospiviroid species are still ubiquitously present in many ornamentals. Since these pospiviroids can be equally harmful to crops as the two quarantine pests PSTVd and CSVd, there is still a risk that transmission occurs from symptomless-infected ornamental plants to economically important crops in Belgium such as tomato, pepper and chrysanthemum.     

Plastids of Marine Phytoplankton Produce Bioactive Pigments and Lipids

Phytoplankton is acknowledged to be a very diverse source of bioactive molecules. These compounds play physiological roles that allow cells to deal with changes of the environmental constrains. For example, the diversity of light harvesting pigments allows efficient photosynthesis at different depths in the seawater column. Identically, lipid composition of cell membranes can vary according to environmental factors. This, together with the heterogenous evolutionary origin of taxa, makes the chemical diversity of phytoplankton compounds much larger than in terrestrial plants. This contribution is dedicated to pigments and lipids synthesized within or from plastids/photosynthetic membranes. It starts with a short review of cyanobacteria and microalgae phylogeny. Then the bioactivity of pigments and lipids (anti-oxidant, anti-inflammatory, anti-mutagenic, anti-cancer, anti-obesity, anti-allergic activities, and cardio- neuro-, hepato- and photoprotective effects), alone or in combination, is detailed. To increase the cellular production of bioactive compounds, specific culture conditions may be applied (e.g., high light intensity, nitrogen starvation). Regardless of the progress made in blue biotechnologies, the production of bioactive compounds is still limited. However, some examples of large scale production are given, and perspectives are suggested in the final section.     

Declining birds in Australian agricultural landscapes may benefit from aspects of the European agri-environment model

Temperate Australia’s wheat/sheep zone and much of Western Europe have both experienced dramatic declines in native bird populations associated with agricultural landscapes. We compare recent conservation strategies on private land in the context of each region’s historical agricultural development and the ecology of its bird fauna. Specifically, we consider which aspects of the conservation instruments and practices employed in European agricultural landscapes might be used to augment and inform approaches to private-land biodiversity conservation in Australia. Australian biodiversity conservation activities have focussed predominantly on remnant native vegetation and rarely target the agricultural matrix (i.e. land that is primarily used for agricultural production). However, declining species include those that not only primarily inhabit woodland, but also species for which components of the agricultural matrix are important, or even their main, habitat. In contrast, in Europe a range of conservation activities undertaken through agri-environment schemes focus explicitly on the management of the agricultural matrix. Whilst the different approaches to conservation on private land in Australia and Europe reflect the two continents’ different ecologies, land-use histories and political economies of agriculture, there are a number of parallels between bird population declines in the two regions, and an opportunity may exist to incorporate some of the successful aspects of the European agri-environment approach into emerging stewardship schemes in Australia. We suggest that the long-term nature of European agri-environment agreements, the principle of landholder payments more commensurate with reduced production opportunity and management actions specifically targeted at the agricultural matrix, are features of the European scheme that could benefit both woodland- and matrix-inhabiting bird species in Australian agricultural landscapes.     

Phthalate Fate in the Hydrographic Network of the River Seine Basin (France) Under Contrasted Hydrological Conditions

Water, Air, &; Soil Pollution - This study investigated how forest soil acidification is affected by edge proximity. We measured pH(KCl) and exchangeable K, Ca, Mg and Al concentrations of the...     

Swine infection with Trichinella spiralis: Comparative analysis of the mucosal intestinal and systemic immune responses

The immune protective response developed by swine against Trichinella spiralis is not yet fully understood, particularly at the mucosal level. This study aimed to characterise intestinal immunity to T. spiralis by comparison with the systemic response in specifically pathogen-free pigs. For this purpose, the kinetics of cytokine and antibody production were assessed in the intestinal mucosa and serum of swine infected with T. spiralis for up to 60 days post-infection (dpi). An ex vivo model of jejunum mucosa culture was used to collect the supernatant as a source of antibodies (Abs). Mucosal antibodies were observed by Western blot from 15 dpi, while serum antibodies were expressed from 20 dpi. Both sources of antibodies initially recognized a 110 kDa protein, followed by the identification of 35, 43/46 and 55/59 kDa proteins. IgG1 and IgA antibodies were strongly expressed within the mucosa. The expression levels of Type 1 (IFN-gamma, IL-12), Type 2 (IL-4, IL-6), pro-inflammatory (TNF-alpha) and regulatory (IL-10, TGF-beta) cytokines were assessed by RT-PCR in the intestinal mucosa and spleen. Both IL-10 and IFN-gamma mRNA levels were increased in mucosa, whereas IL-6 and IL-12 mRNA were expressed in spleen. Taken together, these results demonstrated a mixed Type 1/Type 2 profile, the Type 2 profile being dominant in the mucosa.     

Genetic engineering of plant volatile terpenoids: effects on a herbivore,a predator and a parasitoid

BACKGROUND: Most insect‐resistant transgenic crops employ toxins to control pests. A novel approach is to enhance the effectiveness of natural enemies by genetic engineering of the biosynthesis of volatile organic compounds (VOCs). Before the commercialisation of such transgenic plants can be pursued, detailed fundamental studies of their effects on herbivores and their natural enemies are necessary. The linalool/nerolidol synthase gene FaNES1 was constitutively expressed from strawberry in three Arabidopsis thaliana accessions, and the behaviour of the aphid Brevicoryne brassicae L., the parasitoid Diaeretiella rapae McIntosh and the predator Episyrphus balteatus de Geer was studied. RESULTS: Transgenic FaNES1‐expressing plants emitted (E)‐nerolidol and larger amounts of (E)‐DMNT and linalool. Brevicoryne brassicae was repelled by the transgenic lines of two of the accessions, whereas its performance was not affected. Diaeretiella rapae preferred aphid‐infested transgenic plants over aphid‐infested wild‐type plants for two of the accessions. In contrast, female E. balteatus predators did not differentiate between aphid‐infested transgenic or wild‐type plants. CONCLUSION: The results indicate that the genetic engineering of plants to modify their emission of VOCs holds considerable promise for facilitating biological control of herbivores. Validation for crop plants is a necessary next step to assess the usefulness of modified volatile emission in integrated pest management. Copyright © 2012 Society of Chemical Industry