Baculoviruses as vectors for foreign gene expression in insect cells

Recombinant DNA technology has enabled the expression of a large number of eukaryotic genes in vitro. There has been growing interest in recent years in the development of baculovirus expression vectors as an easily manipulated gene expression system. Both virus and insect cell culture are relatively easy to handle, and biologically active proteins have been produced abundantly from a variety of eukaryotic, prokaryotic and viral genes. This review describes the molecular biology of baculoviruses and some of the current applications of the baculovirus expression system in insect cells. Specific emphasis is placed on those features relevant to the use of this system in pesticide research.     

Further observations on the enhanced degradation of iprodione and vinclozolin in soil

The effects of soil pH on rates of degradation of iprodione and vinclozolin were measured in a silty clay loam soil. Little degradation of either fungicide occurred at pH 4.3 or 5.0, and degradation at pH 5.7 was slower than at pH 6.5. In both of the higher-pH soils, the rate of loss of a second application of either fungicide was faster than that of the first, and a third application degraded even more quickly. In soil with pH 6.5, for example, the times for 50% degradation of iprodione following the first, second and third applications were about 30, 12 and 4 days, and for vinclozolin were 30, 22 and 7 days respectively. Iprodione degraded very rapidly in a sandy loam that had been treated three times previously with this fungicide and also degraded rapidly in the same soil pretreated three times with vinclozolin. Vinclozolin degraded rapidly in the vinclozolin pre-treated soil, but its rate of loss in the iprodione pre-treated soil was only slightly faster than in the previously untreated control. Studies of iprodione degradation in 33 soils from commercial fields demonstrated a clear trend towards faster rates of loss in soils with an extensive history of iprodione use. The time for 90% loss from previously untreated soils varied from 22 to 93 days. It varied from 16 to 28 days in soils treated once previously and from 5.2 to 23 days in soils treated twice previously. In soils that had received three or more previous doses, the time to 90% degradation varied from 3.8 to 15 days.     

The production of azadirachtin by in-vitro tissue cultures of neem,Azadirachta indica

Callus produced from leaves of a Ghanaian strain of the neem tree, Azadirachta indica A. Juss has been shown to produce the natural insecticide azadirachtin when grown in a defined medium. The azadirachtin was isolated by standard procedures of solvent partition and column chromatography monitored by supercritical fluid chromatography. Biological activity was monitored with antifeedant tests using the desert locust (Schistocerca gregaria Forsk.). The azadirachtin was identified by chromatography on three independent chromatographic systems (SFC, HPLC & TLC) and two thin-layer colour tests. It has 100% antifeedant activity at < 0.04mg litre^?1. The yield of azadirachtin was 0.0007% based on dry weight of callus.     

Secreted phosphatase activities in trypanosomatid parasites of plants modulated by platelet-activating factor

ABSTRACT The secreted phosphatase activities of two trypanosomatid parasites were characterized and compared with supernatants of living cells. The plant parasite Phytomonas fran?ai and the phytophagous hemipteran parasite Herpetomonas sp. hydrolyzed p-nitrophenylphosphate at a rate of 15.54 and 6.51 nmol Pi/mg of protein per min, respectively. Sodium orthovanadate (N(a)VO(3)) and sodium fluoride (NaF) decreased the phosphatase activities. The phosphatase activity of P. fran?ai was drastically diminished (73% inhibition) in the presence of sodium tartrate, whereas the phosphatase activity of Herpetomonas sp. was inhibited by 23%. Cytochemical analysis showed the localization of these enzymes on the external surface and in the flagellar pocket of the two trypanosomatids. Sodium tartrate inhibited this reaction, confirming the biochemical data. Platelet-activating factor modulated the phosphatase activities, inhibiting P. fran?ai activity and stimulating Herpetomonas sp. phosphatase activity.     

Microbiological and Chemical Analysis of Neem (Azadirachta indica) Extracts: New Data on Antimicrobial Activity

The antimicrobial effects of extracts of neem seed (Azadirachta indica A. Juss.) were investigated using microbial growth inhibition assays. A laboratory-prepared neem seed extract along with a commercially available formulated product, were characterized using HPLC, and shown to be effective against a range of bacteria in an agar diffusion assay. The active ingredient,i.e., the unformulated seed extract of the commercial product, also showed activity and this was further investigated in a biochromatogram, using the sensitive bacteriumBacillus mycoides. Results showed antibacterial activity as three discrete inhibition zones that did not correspond to the R_f of the major neem metabolites, azadirachtin, nimbin and salannin. This suggests that these compounds were not antibacterial. The colony radial growth rates of the fungal pathogens that cause ‘take-all’ and ‘snow mould’ disease were both significantly affected when the commercial, unformulated, neem seed extract was incorporated into the growth medium. Experiments in liquid culture suggested that the effect was fungistatic. Conidial germination of the commercially important obligate pathogenSphaerotheca fuliginea (powdery mildew) was reduced to 11%. The results show that neem seed extracts possess antimicrobial activity with notable effects on some fungal phytopathogens. This Work demonstrates that neem seed extracts have potential for controlling both microbial and insect pests. http://www.phytoparasitica.org posting Sept. 16,2001.     

Variation of pesticide concentration in sheep dips operated according to traditional and revised methods

OBJECTIVE: To quantify stripping in traditional dipping operations and to revise dipping methods, based on prediction of stripping so that a more stable concentration of pesticide in the dipwash is achieved. DESIGN AND METHODS: Plunge and shower dips were operated sequentially according to traditional and revised dipping instructions. Dips were operated by continuous and intermittent replenishment. Samples of mixed dipwash were collected periodically and assayed for pesticide (diazinon) concentration. RESULTS: Diagrammatic representations of pesticide concentration versus number of sheep dipped indicated traditional dipping leads to wide variations in the concentration of pesticide in dipwash during dipping. Intermittent replenishment led to a 'saw-tooth' pattern in the pesticide concentration. Traditional continuous replenishment (using the starting concentration of pesticide) indicated both the rate and extent of stripping was higher in shower dipping. If sufficient sheep were dipped, equilibrium was reached between the rate of pesticide replenishment and removal. An alternative method of dip operation by continuous replenishment, using a low starting concentration of pesticide and a replenishment concentration high enough to offset the pesticide loss through stripping resulted in a more stable concentration of pesticide in the dip. CONCLUSION: Revision of dipping instructions can lead to exposure of sheep to stable concentrations of stripping pesticide during dipping.