Determination of total phenolic and flavonoid contents in selected fruits and vegetables, as well as their stimulatory effects on mouse splenocyte proliferation

Antioxidant activity index (AAI) by the 2,2-diphenyl-1-picrylhydrazyl method

Honey samples from the seven most common honey types in Slovenia were screened for total phenolic content by the modified Folin– Ciocalteu method, for potential antioxidant activity using the ferric reducing antioxidant power (FRAP) assay and by the 1,1-diphenyl-2picrylhydrazyl (DPPH) method for antiradical activity. In addition the colour characteristics of honey samples were analysed. The results of the study showed that total phenolic content, antioxidant activity and colour parameters differ widely among different honey types. Phenolic content expressed as gallic acid equivalent ranged from 44.8 mg/kg in acacia honey to 241.4 mg/kg in fir honey. Antioxidant activity was the lowest in the brightest acacia and lime honeys and the highest in darker honeys, namely fir, spruce and forest. The colour of the Slovenian honeys, analysed in this study was very variable and ranged from pale yellow to dark brown. Correlations between the parameters analysed were found to be statistically significant (p < 0.05). 2007 Elsevier Ltd. All rights reserved.

Analytical, Nutritional and Clinical Methods Evaluation of the phenolic content, antioxidant activity and colour of Slovenian honey

Evaluation of the phenolic content, antioxidant activity and colour of Slovenian honey

Chemical composition of white (Morus alba), red (Morus rubra) and black (Morus nigra) mulberry fruits

Identification of an antigenic peptide specific for bluetongue virus using phage display expression of NS1 sequences.

DUPLESSIS, DH , VAN WYNGAARDT, W, ROMITO, M , DU PLESSIS, M & MAREE, S 1999 The use of chicken lgY in a double antibody sandwich ELISA fo r detecting African horsesickness virus Onderstepoort Journal of Veterinary Research, 66:25-28 An indirect sandwich ELISA that can detect as little as 8 ng of African horsesickness virus (AHSV) was developed Viral antigen was captured from suspension using an immobilized monoclonal anti­ body specific for an ep itope on VP?, a protein that is a major constituent of the virus core Egg-yolk derived chicken lgY directed against AHSV (serotype 3) was used as the secondary antibody Since lgY and mouse lgG do not cross-react serologically, the secondary antibody was not labell ed, but was instead detected with enzyme-coupled sheep antibodies directed against avian immunoglobulins The assay recognized all nine AHSV serotypes, but not the Cascara isolate of equine encephalosis virus, a related orbivirus that also infects horses In addition to being ab le to detect and quanti fy whole AHSV, the ELISA could show the presence of VP? produced by recombinant baculoviruses

The use of chicken IgY in a double antibody sandwich ELISA for detecting African horsesickness virus.

Immunization with naked DNA was used to elicit chicken egg yolk antibodies (IgY). Layer hens were inoculated with plasmid DNA encoding the enhanced green fluorescent protein, the fusion protein of Newcastle disease virus, and VP2 of African horse sickness virus. IgY was extracted from egg yolks by polyethylene glycol precipitation. Specific antibodies were present in the yolks of eggs from hens immunized with each of the three different plasmids. This approach to raising polyclonal antibodies obviates the need to produce and purify large quantities of proteins for immunization and can potentially yield large amounts of diagnostically or therapeutically useful reagents.

https://www.future-science.com/doi/pdf/10.2144/01313dd05

Eliciting Antigen-Specific Egg-Yolk IgY with Naked DNA

Several field isolates of fowlpoxvirus (FPV) from Burkina Faso, West Africa, were isolated and partly evaluated by molecular analysis. In addition, the in ovo antiviral activity against FPV of a gall extract from Guiera senegalensis was determined. Three viral isolates were obtained from suspected fowlpox cases after passage in embryonating chicken eggs and their poxviral identity confirmed by electron microscopy. All isolates were found to be pathogenic for chicks and all grew well in cell culture. Polymerase chain reaction and sequencing of amplicons revealed sequences identical with those of other FPV strains. The most studied isolate was then employed for use in an antiviral assay. An aqueous acetone extract from the galls of G. senegalensis was found to inhibit both virus-induced pock formation and to reduce viral titre in embryonating chicken eggs. The suggested mechanism of action is the activation of the alternative complement pathway and the inhibition of FPV-induced cholesterogenesis in ovo by constituents of the gall extract.

In ovo inhibition of fowlpoxvirus replication by a gall extract from Guiera senegalensis.

The detection and characterization of specific nucleic acids of medico-veterinary pathogens have proven invaluable for diagnostic purposes. Apart from hybridization and sequencing techniques, polymerase chain reaction (PCR) and numerous other methods have contributed significantly to this process. The integration of amplification and signal detection systems, including on-line real-time devices, have increased speed and sensitivity and greatly facilitated the quantification of target nucleic acids. They have also allowed for sequence characterization using melting or hybridization curves. Rugged portable real-time instruments for field use and robotic devices for processing samples are already available commercially. Various stem-loop DNA probes have been designed to have greater specificity for target recognition during real-time PCR. Various DNA fingerprinting techniques or post amplification sequencing are used to type pathogenic strains. Characterization according to DNA sequence is becoming more readily available as automated sequencers become more widely used. Reverse hybridization and to a greater degree DNA micro-arrays, are being used for genotyping related organisms and can allow for the detection of a large variety of different pathogens simultaneously. Non-radioactive labelling of DNA, especially using fluorophores and the principles of fluorescence resonance energy transfer, is now widely used, especially in real-time detection devices. Other detection methods include the use of surface plasmon resonance and MALDI-TOF mass spectrometry. In addition to these technological advances, contributions by bioinformatics and the description of unique signatures of DNA sequences from pathogens will contribute to the development of further assays for monitoring presence of pathogens. An important goal will be the development of robust devices capable of sensitively and specifically detecting a broad spectrum of pathogens that will be applicable for point-of-care use. Advances in biosensors, the development of integrated systems, such as lab-on-a-chip devices, and enhanced communications systems are likely to play significant future roles in allowing for rapid therapeutic and management strategies to deal with disease outbreaks.

http://centaur.vri.cz/news/prilohy/pril406.pdf

Marco Romito

Papers

Identification of an antigenic peptide specific for bluetongue virus using phage display expression of NS1 sequences.

The use of chicken IgY in a double antibody sandwich ELISA for detecting African horsesickness virus.

Eliciting Antigen-Specific Egg-Yolk IgY with Naked DNA

In ovo inhibition of fowlpoxvirus replication by a gall extract from Guiera senegalensis.

Current and Future Developments in Nucleic Acid-Based Diagnostics