A streak disease of pearl millet caused by a leafhopper-transmitted geminivirus

The cause of a streak disease of pearl millet (Pennisetum glaucum), originating from Nigeria, has been attributed to a geminivirus belonging to the African streak virus cluster. A full-length, infectious clone of the virus was obtained which was transmissible by the vectorCicadulina mbila (Naudé). Analysis of the complete nucleotide sequence of the coat protein gene of this virus shows it to be most closely related to sugarcane streak virus. The possible evolutionary implications of this finding are discussed.     

Characterisation of a maize-infecting potyvirus from Spain

In order to characterise and classify an unknown maize-infecting potyvirus isolated from fields in northeast Spain, the entire coat protein gene and the C-terminal twothirds of the large nuclear inclusion protein (NIb) gene were cloned and sequenced. Protein sequencing enabled the cleavage site between the two proteins to be deduced and also revealed that on storage the viral coat protein undergoes a specific degradation in which the N-terminal 39 amino acids are removed. Comparison of the nucleotide sequence of the 3 non-coding region of the viral RNA and the predicted amino acid sequence of the coat protein with the equivalent regions of other members of the potyvirus group revealed that the Spanish virus is closely related to maize dwarf mosaic virus strain A.     

Chylothorax associated with cryptococcal mediastinal granuloma in a cat

A 10-year-old neutered female cat had chylothorax, precaval syndrome, and a mediastinal granuloma resulting from infection with Cryptococcus neoformans. Diagnosis of a chylous effusion was made by cytologic examination of pleural fluid and by finding higher triglyceride levels in the effusion than in serum (825 vs. 64 mg/dl, respectively). Postmortem examination revealed cryptococcal organisms in the mediastinal granuloma, lungs, cerebral meninges, and connective tissues adjacent to the thyroid gland. Chylous effusion in a cat associated with cryptococcosis has not been reported previously. Cryptococcosis should be included in the differential diagnosis in chylous effusions in cats.     

Antimicrobial substances and chick growth promotion: The growth‐promoting activities of antimicrobial substances,including fifty‐two used either in therapy or as dietary additives

1. Fifty‐five antimicrobial substances were tested for their ability to promote growth when added to the diet of chicks.

2. Both cephalosporins and all the nine penicillins tested were active.

3. Of six aminoglycosides, streptomycin and gentamicin had the greatest activity and neomycin had none.

4. Growth rate was significantly improved by clindamycin, lincomycin, vancomycin, spectinomycin, rifampicin, oxytetracycline, chlortetracycline, erythromycin, tylosin, flavomycin, virginiamycin and zinc bacitracin. Chloramphenicol and nalidixic acid were inactive. Polymixin B, novo‐biocin, cycloserine, phosphonomycin, and sodium fusidate had little activity. Fusidic acid promoted growth at 250 mg/kg diet.

5. Trimethoprim was inactive alone and in combination with sulpha‐diazine. Of seven 5‐nitroimidazoles, only dimetridazole and metronidazole showed slight activity. Of the six 5‐nitrofurans, only nitrovin, the standard reference substance used, promoted growth.

6. Caprylohydroxamic acid, a urease inhibitor, had no beneficial effect on growth rate or on the efficiency of food conversion.

7. The growth‐promoting properties of the various substances could not be related with their known antimicrobial and absorption characteristics in mammals.     

Antimicrobial substances and chick growth promotion: Comparative studies on selected compounds in vitro and in vivo

1. Antibacterial activity of selected compounds in vitro and their ability to promote growth when added to the diet of chicks were not correlated.

2. The difference in growth‐promoting activity between streptomycin, which was active, and kanamycin which was not, was not related to their effects on the flora adhering to the wall of the crop, jejunum or ileum, to their relative toxicity to the chick nor to differences in their stability in the diet.

3. When low concentrations ofkanamycin or streptomycin were given, the antibiotic was concentrated in the caecal contents. In contrast, penicillin was undetectable in the caeca of birds given high concentrations of benzyl penicillin.

4. In chicks given benzyl penicillin, the numbers of Streptococcus faecium increased at the expense of Strep, faecalis. This probably resulted from changes in the crop flora.

5. Birds given high dietary concentrations of benzyl penicillin showed improved weight gains, in spite of increased numbers of Escherichia coli in the small intestine.     

‘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.     

Development of an EU protocol for the detection and diagnosis of Potato spindle tuber pospiviroid*

The work described here formed part of the EU SMT DIAGPRO project, to develop diagnostic protocols for 18 regulated pests. The Potato spindle tuber pospiviroid (PSTVd) protocol was developed primarily for testing in vitro‐ and glasshouse‐grown potato plants for the purposes of post‐entry quarantine and the production of pathogen‐tested nuclear stock. After a performance audit of methods used by 12 laboratories in Europe and America by ring testing, four methods were chosen for multilaboratory validation. For most laboratories, the detection limits were 10–20 mg of PSTVd‐infective tissue for R‐PAGE; 0.25–0.5 mg for DIG‐probe; 0.062 mg for RT‐PCR; and 0.0155 mg for TaqMan (this was the lowest weight of infective tissue tested). Some laboratories were able to extend the detection limit to 0.0155 mg for DIG‐probe and RT‐PCR. The DIG‐probe and R‐PAGE are recommended as primary detection methods, with confirmation of viroid presence by any of the four validated detection methods. Specific diagnosis requires the viroid to be sequenced. Other methods may be used for primary detection, providing that they preferably detect all PSTVd isolates and other Pospiviroids that have the potential to infect potato, and detect viroid in at least 1/10 of the tissue weight normally tested per plant.     

How to perform transvenous electrical cardioversion in horses with atrial fibrillation

Electrical cardioversion of atrial fibrillation is a well-established technique for restoration of sinus rhythm in humans. While transthoracic cardioversion is more commonly used, transvenous electrical cardioversion (TVEC) has been reported as having higher efficacy at substantially lower energy levels. In horses, treatment of atrial fibrillation has essentially been limited to the administration of quinidine salts either orally or intravenously. TVEC provides an alternative to quinidine salts, especially for those animals in which quinidine is neither effective nor tolerated. The present report details this technique in horses, discusses possible complications of the procedure, and provides guidance for successful outcome. Still and video images are used to illustrate details with regard to TVEC techniques in horses. Please view supplemental material for the videos.