Endotoxin absorption in hay-fed and lactic acidotic sheep.

Absorption of endotoxin from the gastrointestinal tract was evaluated in hay-fed and lactic acidotic sheep duodenally infused with 10 mg of Escherichia coli endotoxin, and in lactic acidotic sheep not infused. The effect of abomasal fluid on biological activity of endotoxin was also evaluated. Leukopenia was the criterion used for detecting endotoxemia. Absorption of endotoxin from the gastrointestinal tract was not detected in either hay-fed or lactic acidotic sheep. Endotoxin appeared to maintain its activity after incubation with abomasal fluid, and the presence of endogenous endotoxin in abomasal contents was indicated. The results indicate that endotoxin of alimentary origin may not be involved in the lactic acidosis syndrome in ruminants.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

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 (V_L) and heavy chain (V_H) 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.     

The effects of progesterone on oviposition and ovulation in the domestic fowl (Gallus domesticus)

1. Following an injection of 0.5 or 0.1 mg progesterone/kg between 0 and 6 h after ovulation, oviposition of the resulting egg was delayed by 1 to 11 h and occurred 26 to 31 h after injection, depending on the dose. The injection terminated the laying of a sequence of eggs by causing the next ovulation to occur a day late. The delayed ovulation occurred at the time normally expected for the first ovulation of a sequence and became the first of a new sequence.

2. Following an injection of 0.5 or (H mg progesterone/kg between 6 and 15 h after ovulation, oviposition of the resulting egg was generally delayed by between 15 and 28 h and occurred at the same time of day as the next ovulation, which was delayed as in the first experimental situation. Subsequent ovulations were resynchronised and followed at intervals according to the normal sequence established before the injection.

3. Injection of 0.5, 0.1 or 0.05 mg progesterone/kg between 12 and 9 h before an expected ovulation advanced the oviposition of the egg already in the uterus (shell gland) by about 3 h. The succeeding ovulation was either advanced or blocked.

4. These observations suggest that the pre‐ovulatory surge of progesterone is directly or indirectly involved in the timing of oviposition and ovulation.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

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     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

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     

Combined effect of generally regarded as safe (GRAS) compounds andTrichoderma harzianum on the control of postharvest diseases of rambutan

Botryodiplodia theobromae, Colletotrichum gloeosporioides andGliocephalotrichum microchlamydosporum are the causal fungi of the rambutan postharvest diseases stem-end rot, anthracnose and brown spot, respectively. Two different treatments of rambutan fruits(Nephelium lappaceum) against the three pathogens were compared: potassium metabisulphite (250 ppm) or cinnamaldehyde (30 ppm), each combined withTrichoderma harzianum (TrH 40). The application of TrH 40 and potassium metabisulphite effectively controlled the incidence and severity of the three postharvest diseases and maintained the overall quality and color of the fruit under low temperature storage at 13.5°C and 95% r.h. for 18 days. The greatest effect of this treatment was shown onG. microchlamydosporum. Cinnamaldehyde affected the growth and germination of TrH 40, whereas potassium metabisulphite did not. http://www.phytoparasitica.org posting Nov. 4, 2001.