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Prolamin content of buckwheat flour and processed foods was 24.2–42.1 mg/kg dry material measured by ELISA. According to in vitro results buckwheat is suitable for use in coeliac diet, although it contains some antinutritive materials, protease inhibitors and tannin. The allergenic properties of buckwheat are poorly understood. In our investigation intensity of the 24 kD protein band of buckwheat, of which allergenic activity is known has decreased, and 30–35 kD protein associations have been formed after heat treatment. Immunochemical reaction of buckwheat proteins were studied with blood specimens of coeliac and healthy persons.
Colmenares, R. (1998): Proposal for the ELISA Internet Conference: Biodiversity Area (ELISA Interim Report), ECNC, Tilburg. Colmenares R
Bever Mike Crowley Chris S Duvall Scott Fitzpatrick Elisa Guerra Marco Leonti Mark Merlin Jeffrey Peytrequin Gomez David Presti Carl Ruck Giorgio Samorini Miklós Sárközy Theodore
Summary
In the present work, we investigated the immunological behavior of bothropstoxin-1, a K49 phospholipase from Bothrops jararacussu, and of ovalbumin before and after irradiation with 60Co g-rays. Isogenic mice were immunized with either native or irradiated proteins. The circulating antibodies were isotyped and titrated by ELISA. Results indicate that irradiated proteins were immunogenic and the antibodies elicited by them were able to recognize the native proteins in ELISA. Data also indicate that the irradiated protein induced higher titers of IgG2a and IgG2b, suggesting that Th1 cells were predominantly involved in the immune response. Structural modifications of the proteins were investigated by SDS-PAGE, mass spectrometry and size exclusion chromatography. According to our data, irradiation promoted structural modifications on both proteins, characterized by higher molecular weight forms (aggregates and oligomers). When analyzed by mass spectrometry, the irradiated bothropstoxin appeared in several oxidized forms. These results indicate that irradiation of toxic proteins can promote significant modifications in their structures, but still retain many of the original antigenic and immunological properties of native form.
A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5´-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1h group.
Lyme borreliosis, granulocytic anaplasmosis and monocytic ehrlichiosis are well studied in humans and dogs. In horses, these diseases are not widely investigated and limited information is available about their occurrence. The purpose of this study was to present the first ELISA-based report on the seroprevalence of Anaplasma phagocytophilum, Ehrlichia spp. and Borrelia burgdorferi in horses from Northern Bulgaria. A total of 192 horses were investigated from three regions in Northern Bulgaria (Northwestern, North-Central and Northeastern Bulgaria). All equine sera were tested for A. phagocytophilum, Ehrlichia spp. and B. burgdorferi antibodies by a commercial rapid ELISA test. Antibodies against A. phagocytophilum were found in all the three regions at a mean frequency of 12% (23/192), ranging from 9.38 to 15.63% by region. Antibodies against Ehrlichia spp. were found in horses from one region (Northeastern) at a rate of 0.5% (1/192). Anti-B. burgdorferi antibodies were detected in all the three regions with a mean frequency of 15.1% (29/192), ranging from 14.06 to 17.19% by region. A co-exposure to A. phagocytophilum and B. burgdorferi was observed in 6.3% of the cases (12/192). This is the first report on the natural exposure of horses to these bacteria (A. phagocytophilum, Ehrlichia spp. and B. burgdorferi) in Northern Bulgaria.
Simple real-time PCR assay with one set of primer and probe for rapid, sensitive qualitative and quantitative detection of Entamoeba histolytica has been used. Consensus sequences were used to amplify a species-specific region of the 16S rRNA gene, and fluorescence resonance energy transfer hybridization probes were used for detection in a LightCycler platform (Roche). The anchor probe sequence was designed to be a perfect match for the 16S rRNA gene of Entamoeba species, while the acceptor probe sequence was designed for Entamoeba histolytica, which allowed differentiation. The performed characteristics of the real-time PCR assay were compared with ELISA antigen and microscopical detection from 77 samples of individuals with suspected clinical diagnosis of imported E. histolytica infection. Stool and liver abscess pus samples were examined with analytical sensitivity of 5 parasites per PCR reaction. The melting curve means Tms (standard deviation) in clinical isolates were 54°C. The real-time assay was 100% sensitive and specific for differentiation of Entamoeba histolytica, compared with conventional ELISA or microscopy. This real-time PCR assay with melting curve analysis is rapid, and specific for the detection and differentiation of Entamoeba histolytica. The suitability for routine use of this assay in clinical diagnostic laboratories is discussed.
The aims of the present study were to determine (a) the effectiveness of an attenuated live Cryptobia salmositica vaccine; (b) the effects of food deprivation on the immune response and its duration in rainbow trout ( Oncorhynchus mykiss ) immunised with a live C. salmositica vaccine or with a killed Aeromonas salmonicida vaccine. The fish were divided into three groups (I, II and III; 14 fish per group), those in Groups I and II were under food deprivation (0.40% of body weight), while Group III fish were fed to satiety. The study showed that the attenuated strain of C. salmositica did not cause anaemia and disease, and the fish were protected from clinical disease when they were challenged with virulent parasites. Parasitaemia in all fish vaccinated and challenged with virulent C. salmositica fluctuated and was relatively low; however, fish in Group III had higher parasitaemia than those in Groups I and II between weeks 8 and 14. The numbers of activated neutrophils increased [nitroblue tetrazolium (NBT) assay] after immunisation with both Cryptobia and Aeromonas vaccines and they remained high throughout the experiment. Antibody production (ELISA values) increased after vaccination and were slightly higher in Group III. ELISA titres against A. salmonicida increased after vaccination and decreased after 5 weeks. The titres increased again after the vaccinated fish were given booster, and they were higher than those in the first vaccinated fish.
Grapevine fanleaf virus (GFLV) is the causal agent of a widespread disease that affects vineyards. Since it is difficult to culture viruses, the availability of an easy and efficient method of virus maintenance in the laboratory would be of interest to virologists. The objective of this research was to determine an adequate culture medium that promotes callus growth and permits the preservation of GFLV on Vitis vinifera tissue. Fragments of in vitro cultured leaves (25 mm 2 ), originated from Cabernet Sauvignon positive for GFLV, were cultivated on a Murashige and Skoog (1962) medium, and the callus was monitored for the presence of GFLV every two weeks using ELISA. Higher 2,4-D concentration induced a higher growth, particularly when combined with a low BA concentration. The medium enriched with 1.0 ppm of 2,4-D combined with 0.5 ppm of BA showed the best result, with the callus area reaching more than 250 mm 2 after 8 weeks in culture. ELISA absorbance observed on callus tissues during the whole period was, at least, three times higher than that observed on leaves positive for GFLV kept either in vivo or in vitro and more than 18 times higher than that of the negative control. Any remarkable difference in absorbance was recorded during the period of callus cultivation. It was concluded that the viral load on the callus was not affected during this time, suggesting that this kind of in vitro culture is an efficient method to preserve GFLV.
Plants of wild species Sinapis arvensis L . — (wild mustard) with severe mosaic symptoms were established all around different farm crops in Sofia valley and even in the Sofia suburbs in 2006. Two viruses were identified by ELISA method (DAS-ELISA) and by the indicator method. Those were the virus of the genus Caulimovirus , family Caulimoviridae — Cauliflower mosaic virus (CaMV) and a virus of the genus Potyvirus , family Potyviridae — Turnip mosaic virus (TuMV). Both viruses were found in wild mustard plants, often in mixed infection. Turnip yellow mosaic virus (TYMV) wasn’t determined in wild mustard plants. CaMV was identified by the infection of its diagnostic indicator species Brassica oleracea var. botrytis (cv. Snowball) — cauliflower. Besides cauliflower, TuMV was also identified in cabbage — Brassica oleracea var. capitata, cv. Balkan, Chenopodium amaranticolor, Chenopodium quinoa and tobacco plants — Nicotiana rustica and Nicotiana tabacum , cv. Samsun. The economically important crop oilseed rape Brassica narus var. oleifera , hybrid Elvis, was infected by artificial mechanic inoculation with material from diseased wild mustard. Oilseed rape plants responded with local and systemic chlorotic mottling of the leaves. A clearly pronounced mosaic appeared on leaf petioles of the growth. Our hypothesis was that the weed Sinapis arvensis could be a reservoir of TuMV and CaMV infection for oilseed rape under natural conditions, if the viruses are transferred from wild mustard to oilseed rape with aphids.
