水稻白叶枯病杂草带菌和传病的研究

水稻白叶枯病自发现到现在已有90年的历史。由于发生地区广泛,为害严重,各国对此病曾做了大量研究。日本对它已研究了半个多世纪,我国也有二十多年的研究经历。但对其侵染来源和传播途径,仍有不同看法。我国最初认为种子传病是首要的。但在病区单抓种子处理还不能收到预期效果,又认为病稻草是病区次年的主要菌源。近年来,又发现在严格处理病革、病种子的情况下,仍然发病,故怀疑除上述两种传播途径外,还有其它传播途径。关于杂草传病问题,日本已进行了多年的研究,但和我国国内一样,对杂草带菌传病问题,意见很不一致。我们认为对这一传播途径有彻底弄清的必要,它不仅关系到一个病的侵染循环问题,而且直接关系到对白叶枯病的彻底防治。如果这一传播途径确实存在,我们若不承认它,在防治上就会出现大的漏洞。如果白叶枯病确与杂草无关,盲目地采用相应的措施,也是多余而毫无意义的。这次试验是采用较直接的“离心浓缩针刺接种法”,其结果:第一年证实有马唐等六种杂草、黄豆和水稻根部能够带菌;第二年证实有车前等三种杂草和水稻根不仅带菌,而且可越冬并存活到次年五月;第三年又证实在不同地区有马唐,狗牙根等四种杂草能够带菌越冬,并存活到次年五月中旬。另外,用病区杂草和水稻健苗混栽,结果稻苗上发病,而对照的不发病。通过三年来的试验研究,证实有数种杂草的根茎部不仅带菌,而且能传病。     

‘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.     

First record of bacillary rickettsia-like organisms in European tick Dermacentor marginatus (Sulzer).

Bacillary rickettsia-like organisms (BRLO) were found in the tick Dermacentor marginatus. They are gram-negative and differ from common bacteria and reckettsiae both in the cultivation conditions and morphology. BRLO are non-pathogenic for ticks and guinea pigs. In our studies they were isolated on half-engorged females of D. marginatus, on which they are still maintained.     

Histological studies on cow intestinal mucosa and electron microscopic studies of the ruminal epithelium of cows fed a deficient diet for a prolonged time]

Histological and histochemical studies were carried out on the gastro-intestinal mucosa of three experimental cows (roughages/maize silage) and three control animals. The animals were slaughtered on termination of the long-term trial. Mucosa samples were taken, for further study, from the rumen, the duodenum, the jejunum, the large intestine and the appendix. Histochemical analysis did not reveal any essential differences in the activities of non-specific esterase, alkaline and acid phosphatase and lactic acid dehydrogenase in the mucosa of the rumen, the large and the small intestine and the appendix of both the experimental animals and the controls. The experimental animals were found to exhibit a higher rate of glutamate-dehydrogenase activity in the ruminal mucosa and in the mucosa of the large and small intestine. A higher succinate dehydrogenase activity was observed in the ruminal mucosa of the experimental animals, relative to that of the controls, while the activity in the intestinal mucosa was decreased. Only slight changes were noted in the activity of the enzymatic systems tested. Electron microscopic studies did not reveal any differences in the ultrastructure of the epithelial cells of the ruminal mucosa in both the experimental animal and the controls.     

Testing of the protective effect of cell-free lyophilized vaccine against Marek's disease in field trials]

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.     

Relevance of the variability of kidding day in Creole goats in Guadeloupe

The results of 5732 records of kids born between 1985 and 1996 at Gardel Agricultural Experiment Station (INRA) in Guadeloupe, were used in order to estimate the effect of kidding day (KD) on individual preweaning growth performances, total productivity of Creole goats and litter size. The flock was subjected to a restricted mating season for a long time, by using male effect. The results of the fixed linear model showed a highly significant (P<0.001) effect of KD on growth rate and total productivity of does. Live weights of kids born around the 21st day of the kidding period (KP) was 4% to 7% higher than those of kids born the first day of the KP. For total productivity of does, this ratio did not reached more than 4%. The optimum at 70 days of age occurred around 14th day of the KD with 3% of improvement of total productivity. No effect was observed upon litter size. The genetic (co)variance components were estimated by six different Individual Animal Models. The heritability (h²) estimated from the best model, was h_D²=0.25±0.05 for genetic direct effect; h_M²=0.09±0.04 for genetic maternal effect and the genetic correlation between direct and maternal effects was −0.86±0.12. The use of KD could be highly recommended in a breeding program in this population of Creole meat goats, since it is quite easy to record under commercial conditions as a character related to reproductive performance.     

Pregnancy Rates in Repeat-breeder Heifers Following Multiple Artificial Inseminations during Spontaneous Oestrus

Hormonal asynchronies during oestrus, related to the presence of suprabasal plasma-progesterone (P4) concentrations and a delayed ovulation, interfere with the fertility of repeat-breeder heifers (RBH). Since tubal dysfunction can occur in connection with hormonal asynchronies and constrained availability of fertile spermatozoa at the time of ovulation, the present study tested the hypothesis that frequent sperm deposition from onset of oestrus to ovulation may improve pregnancy rates in RBH. Five RBH and five virgin heifers (VH; controls) were repeatedly artificially inseminated (AI) at 6 h intervals from onset of oestrus to spontaneous ovulation. Hormone analyses revealed suprabasal P₄ concentrations and a delay in the occurrence of the luteinising hormone (LH) surge, but a normal cortisol profile in RBH. Compared with controls, RBH presented longer interval from onset of oestrus to ovulation, and therefore, received more AIs. Pregnancy rates in RBH reached control levels (60%; NS), indicating that the hypothesis might be correct. Pregnancy rates in VH were below the expected range, presumably attributed to a deleterious influence of the frequent handling. The study suggests that pregnancy rates can be improved in RBH by frequent AI in relation to spontaneous ovulation. However, this practice of repeated manipulations, while seeming not to show adverse effects, lacks practicality for routine use.     

DNA Immunization Against Very Virulent Infectious Bursal Disease Virus with VP2‐4‐3 Gene and Chicken IL‐6 Gene

The present study was to investigate the feasibility and efficiency of the DNA vaccine to protect chickens against very virulent infectious bursal disease virus (vvIBDV) infection. A plasmid DNA carrying VP2‐4‐3 genes of vvIBDV SH95 and a plasmid DNA carrying chicken interleukin‐6 (ChIL‐6) genes were constructed and designated as pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 respectively. Several DNA vaccination experiments were performed: 1‐week‐old chickens were intramuscularly injected with only plasmid pcDNA3‐VP2, pALTER‐MAX‐VP2‐4‐3 or mixture with pALTER‐MAX‐ChIL‐6. The chickens at 4 weeks old were orally inoculated with vvIBDV SH95. The results showed that immunization with the mixture of pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 three times conferred protection for 90% of chickens. Enzyme‐linked immunosorbent assay (ELISA) antibody titres in chickens immunized together with pALTER‐MAX‐ChIL‐6 were higher than those immunized simply with plasmid pcDNA3‐VP2 or pALTER‐MAX‐VP2‐4‐3. IBDV was not detected in the bursa of the protected chickens at 8 days after challenge by RT‐PCR. The results indicate that protection against vvIBDV can be achieved by using the VP2‐4‐3 gene of vvIBDV as a DNA vaccine. Furthermore, the simultaneous injection of ChIL‐6 plasmid significantly increased the protection after challenge with the very virulent strain.