New mite-borne virus isolates from rakkyo,shallot and wild leek species

Flexuous viruses were transmitted from rakkyo (Allium chinense) and wild leek species (especiallyA. commutatum) to plants of crow garlic (A. vineale), by transfer of dry bulb mites. By electron microscope decoration tests using three antisera and by inoculations onto test plants, it was concluded that from each of the two natural host species at least two viruses were isolated. The viruses from wild leeks are both pathogenic onAllium spp. and may be of economic importance. Decoration tests on a virus mixture from shallot obtained earlier, revealed another new mite-borne virus in this species. The mite-borne viruses ofAllium spp. appear to be very common; they are largely diverse and their identification remains difficult.     

Exploration of glycoprotein structures: Sequences and consequences

The vast expansion in our knowledge of the detailed molecular structure of many glycoproteins has been achieved by purpose-designed methodologies. In this review, principal experimental methods for gaining fundamental evidence are assessed with respect to well-known iV-glycoproteins (e.g. ribonucleases), 0,N-glycoproteins (e.g. glycophorin A) and O-glycoproteins (e.g. submaxiilary gland mucins and some sulfated forms). Relevant properties of N-carbohydrate-peptide links (N-acetylglucosaminyl-asparagine) and O-linkages (N-acetylgalac-tosaminyl-Thr (or -Ser)) are reviewed and the classification and structure of more elaborate glycoproteins is outlined. Key reactions, both chemical and enzymic, effect the cleavage of N- and O-oligosaccharide side-chains from core protein, and subsequent calibrated chromatographic analysis can permit the identification of certain oligosaccharide groupings characteristic of glycoforms (families of glycoproteins different only in the oligosaccharide chain lengths and bond structures). Basic sequencing of individual oligosaccharides is generally achieved by chemical methods (e.g. methylation analysis, degradation with periodate) and by the action of well-characterised specific exo- and endoglycosidases. Degradation of a glycoprotein chemically (e.g. by cyanogen bromide) or enzymically by endopeptidases to glycopeptides provides means for locating oligosaccharide position(s) on the core protein. The presence of O-sulfate ester groups on oligosaccharide side-chains presents further challenging problems for sequencing investigations. Some correlations between oligosaccharide sequences and biological function are considered. While, in the main, the best-known glycoprotein structures are for those of mammalian origin (e.g. Igs), evidence points to other areas of interest in plants (especially yeasts), invertebrates and micro-organisms. Allusion is made to current comparative and fundamental physico-chemical techniques, especially gel permeation chromatography, membrane-transfer, FAB-MS and NMR (NOE, COSY, etc.).     

Supercritical fluid chromatography—finding a place in the chromatographers' arsenal?

In the mid- to late 1980s, supercritical fluid chromatography (SFC) was ‘reborn’ as a technique for the 1990s. Supercritical fluids have physical properties (viscosity, diffusivity and density) intermediate between those of licuids and gases. Analyte solubilities are similar to those in some liquid chromatography solvents and the gas-like properties of supercritical fluids make higher resolution or shorter analysis times possible. Applications and experience with a home-built SFC system, as well as methods developed elsewhere, are described. The successes and limitations of the technique are reviewed together with details of the instrumentation. Areas in which SFC may become the procedure of choice include multi-residue methods, non-polar analyses, micro-preparative uses and for difficult extractions. The advances in GC and HPLC technology, however, lead us to believe that SFC will remain a specialist technique, appealing only to a minority of analysts.     

Experimental reproduction of severe disease in CD/CD pigs concurrently infected with type 2 porcine circovirus and porcine reproductive and respiratory syndrome virus

Three-week-old cesarean-derived colostrum-deprived (CD/CD) pigs were inoculated with porcine circovirus type 2 (PCV2, n = 19), porcine reproductive and respiratory syndrome virus (PRRSV, n = 13), concurrent PCV2 and PRRSV (PCV2/PRRSV, n = 17), or a sham inoculum (n = 12) to compare the independent and combined effects of these agents. Necropsies were performed at 7, 10, 14, 21, 35, and 49 days postinoculation (dpi) or when pigs became moribund. By 10 dpi, PCV2/PRRSV-inoculated pigs had severe dyspnea, lethargy, and occasional icterus; after 10 dpi, mortality in this group was 10/11 (91%), and all PCV2/ PRRSV-inoculated pigs were dead by 20 dpi. PCV2-inoculated pigs developed lethargy and sporadic icterus, and 8/19 (42%) developed exudative epidermitis; mortality was 5/19 (26%). PRRSV-inoculated pigs developed dyspnea and mild lethargy that resolved by 28 dpi. Microscopic lesions consistent with postweaning multisystemic wasting syndrome (PMWS) were present in both PCV2- and PCV2/PRRSV-inoculated pigs and included lymphoid depletion, necrotizing hepatitis, mild necrotizing bronchiolitis, and infiltrates of macrophages that occasionally contained basophilic intracytoplasmic inclusion bodies in lymphoid and other tissues. PCV2/ PRRSV-inoculated pigs also had severe proliferative interstitial pneumonia and more consistent hepatic lesions. The most severe lesions contained the greatest number of PCV2 antigen-containing cells. PRRSV-inoculated pigs had moderate proliferative interstitial pneumonia but did not develop bronchiolar or hepatic lesions or lymphoid depletion. All groups remained seronegative to porcine parvovirus. The results indicate that 1) PCV2 coinfection increases the severity of PRRSV-induced interstitial pneumonia in CD/CD pigs and 2) PCV2 but not PRRSV induces the lymphoid depletion, granulomatous inflammation, and necrotizing hepatitis characteristic of PMWS.     

Canine renal pathology associated with grape or raisin ingestion: 10 cases.

Ten dogs suffered acute renal failure after ingesting > or = 3 g/kg (dry matter) of grapes or raisins. All dogs had degeneration or necrosis (or both) of proximal renal tubules with basement membranes remaining intact, and epithelial regeneration was observed in 5 out of 10 cases. Mineralized tubular debris or granular to proteinaceous casts (or both) were present in all cases. A golden-brown, globular, intracellular pigment of varying amounts and sizes was observed in 6 out of 10 cases with variable reaction with Prussian blue. Multifocal fibrinous arteritis of the large colon was seen in 2 out of 5 cases with globulin insudation of vessel wall demonstrated by immunohistochemical staining for immunoglobulin (Ig)G and IgM. Mineral analysis on frozen renal tissue from 2 out of 2 cases revealed mildly elevated Ca:P ratio in both. Clinically significant observations were preservation of the integrity of basement membranes after grape-induced tubular injury and presence of early epithelial regeneration. Thus, recovery may be possible if anuria is aggressively managed. With respect to potential pathophysiologic mechanisms, further research into the roles of calcium homeostasis, vascular reactivity, and the significance of the golden-brown pigment is indicated.     

Kinetic and thermodynaimic aspects of the dye-sensitised photo-oxidation of bioresmethrin and related compounds

The Rose Bengal sensitised photo-oxidation of bioresmethrin (I), 2-benzylfuran-4-ylmethyl acetate (II), and 2-benzylfuran-4-ylmethanol (III) has been studied in methanol and methanol + water (1 + 1 by volume) solutions by monitoring the rate or oxygen removal from the system. The results indicate that the compounds studied are oxidised via reaction with singlet oxygen. Values for the rate constants for reaction of singlet oxygen with compounds I-III have been determined over the temperature range 293–308 K, and the energy of activation and entropy of activation for the reactions have been determined. The data have been interpreted in terms of the site of attack of singlet oxygen on the compounds.     

Detection of Spongospora subterranea using monoclonal antibodies in ELISA

Five hybridoma cell lines secreting antibodies (MAbs) recognizing zoospores of S. subterranea were raised from splenocytes of mice. One MAb also weakly recognized plasmodia/zoosporangia and cystosori of S. subterranea , and another recognized only plasmodia/zoosporangia in plate-trapped antigen ELISA. Polymyxa graminis was recognized most strongly out of 26 micro-organisms other than S. subterranea against which the MAbs were tested. Most were recognized only weakly or not at all. The MAb that recognized zoospores of S. subterranea most strongly detected as few as three zoospores per microtitre plate well when 12 replicate wells per treatment were arranged randomly on plates and absorbance values subjected to analysis of variance. The sensitivity of detection was not improved by mixing antibodies, using a biotin-streptavidin amplification system, or by using a double antibody sandwich system. Zoospores of S. subterranea flushed from soil were detected only after unrealistically large numbers of cystosori had been added. They were not detected in samples of naturally infested soil removed from a field shortly after a severely scabbed potato crop had been harvested.     

Components of resistance to late blight (Phytophthora infestans) in eight South AmericanSolanum species

Four components of partial resistance toPhytophthora infestans were measured after inoculation in the greenhouse and in the field ofSolanum arnezii x hondelmannii, S. berthaultii, S. circaeifolium, S. leptophyes, S. microdontum, S. sparsipilum, S. sucrense andS. vernei, and four hybrid progenies ofS. microdontum withS. tuberosum. The four components were infection efficiency, lesion growth rate, generation time and sporulation capacity. The results were compared with resistance ratings derived from field experiments, and with observations made on the potato cultivars Bintje, Bildstar, Libertas and Pimpernel. Genetic variation for all components was found, while the relative importance of the components of partial resistance appeared to vary between the species. InS. microdontum, generation time, infection efficiency and lesion growth rate, and inS. tuberosum infection efficiency, lesion growth rate and sporulation capacity appeared positively associated, but in other species no such association was found. A strong hypersensitive reaction, the expression of which appeared to depend on environmental conditions, was found inS. microdontum. ForS. berthaultii, infection efficiency appeared to be the main resistance component.Abbreviations ADPC area under the disease progress curve - IE infection efficiency - LGR lesion growth rate - GT generation time - SC sporulation capacity     

Potential of biopesticides in agriculture

All living organisms are subject to predation, parasitism or competition from other organisms. The study of these interactions has led to the identification of many potential opportunities for the use of living organisms as biopesticides to protect agricultural crops against insect pests, fungal, bacterial and viral diseases, weeds, nematodes and mollusc pests. A range of biopesticide products (including as active agents bacteria, fungi, nematodes, protozoa, viruses and beneficial insects) are now available commercially for control of insect pests, fungal and bacterial diseases and weeds. However, world biopesticide sales in 1990 were estimated to be $120 million, representing less than 0–5% of the world agrochemical market. Over 90% of biopesticide sales are represented by a single product type, containing Bacillus thuringiensis Berl., for control of insect pests. Nevertheless, biopesticide sales are estimated to be increasing at 10–25% per annum whilst the world agrochemical market is static or even shrinking. There has been a significant renewal of commercial interest in biopesticides as evidenced by the substantial number of alliances forged between major agrochemical companies and biotechnology companies which allow these major companies access to marketing rights to novel biopesticides. This paper reviews the current commercial status of biopesticides and discusses the technical and commercial constraints which have impeded development of biopesticides in the past. Novel developments in R&D, which may enable some of these constraints to be overcome, are examined by reference to a number of specific examples (some of which arise from the author's own experience in a biotechnology company). The future prospects for biopesticides are discussed in the light of technical advances and commercial and regulatory requirements.