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21.
Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (VL) and heavy chain (VH) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.  相似文献   
22.
Accordingly to the subacute toxicity of chlorphacinone to Japanese quail (LC 50: 60 mg/kg feed), the uptake of Delicia-chlorphacinone-baits for several days can be lethal to birds as a matter of principle. A subchronic study resulted in a "no-observed-effect-level" of 1 mg chlorphacinon/kg feed. Reproduction is not affected until dose levels which resulted in lethality or are accompanied by physiological changes (coagulability of blood, microcytic anaemia). Increased sensibility of descendants to chlorphacinone cannot be derived from the findings.  相似文献   
23.
4 digestibility trials (4 male sheep per group) and 2 growth trials were carried out with 24 (V 1) and 32 (V 2) fattening lambs to investigate the effect of abundant oral Fe doses on the digestibility of crude nutrients in a ration of barley + dried green feed and on the results of fattening and carcass yields. Fe supplementation (less than 1400 mg per sheep/day) did not significantly change the digestibility coefficients of crude nutrients. Increasing Fe levels in the concentrates decreased the rates of liveweight increase and food consumption in both growth trials. Well-established statistical evidence was provided for the decline in absolute carcass yields resulting from the lower weight of the lambs at the end of the fattening period after Fe supplementation.  相似文献   
24.
The protective effect of a lyophilized vaccine against Marek's disease ("Keramvac"--Pfizer), prepared from a cell-free turkey's herpesvirus (strain FC 126), was compared in field trials with losses in the group of non-vaccinated chickens. Under the indicated conditions, the vaccine had only a 50.86% effectiveness. The possible causes of the reduced vaccination effect are discussed with regard to the pathomorphological and virological findings suggesting, among others, an increased incidence of the symptoms of the classical form of Marek's disease in the population investigated.  相似文献   
25.
The effect of true milk use in the diet of Spanish Brown Swiss male calves on animal performance, carcass and meat quality, and fatty acids composition was studied. In experiment 1, the effect of milk intake [ad libitum continuous (ADLIB) feed vs. restricted 0.7 during 75 days followed by ad libitum feed (RESTR)] and slaughter endpoint (225 kg vs. 5 month) were studied. In experiment 2, ad libitum concentrate feeding [grain-fed (GF)] was compared with milk supplementation until slaughter [milk-fed (MF)] in calves slaughtered at 345 kg. As regards to milk intake, carcass weight and degree of fatness were higher in the ADLIB group (P<0.05). The RESTR group revealed a higher percentage of saturated fatty acids (P<0.05). In experiment 2, the MF group exhibited a higher fat percentage (P<0.05), lower press and cooking losses (P<0.05), and higher scores for tenderness and juiciness (P<0.05) than the GF group. The percentage of saturated fatty acids was higher in the MF group (P<0.05). The results suggest that true milk use in veal production could be an advantageous alternative in terms on production costs, animal performance, and carcass and meat quality.  相似文献   
26.
An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.  相似文献   
27.
L11A-Fukushima (L11A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L11A-F and L11A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L11A-F appears to possess the properties necessary for practical use. To manage L11A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L11A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002  相似文献   
28.
DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001  相似文献   
29.
Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.  相似文献   
30.
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