Properties of a unique mutant of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus that exhibits a partial many polyhedra and few polyhedra phenotype on extended serial passaging in suspension cell cultures

Serial passaging of wild-type Helicoverpa armigera, single-nucleocapsid (HaSNPV) in H. zea (HzAMI) illsect Cell Cultures results ill rapid selection for the few polyhedra (FP) phenotype. A unique HaSNPV mutant (ppC19) was isolated through plaque purification that exhibited a partial many polyhedra (MP) and FP phenotype. Oil serial passaging in suspension cell cultures, ppC19 produced fivefold more polyhedra than a typical FP mutant (FP8AS) but threefold less polyhedra than the wild-type virus. Most importantly, the polyhedra of ppC19 exhibited MP-like virion occlusion. Furthermore, ppC19 produced the same amount of budded virus (BV) as the FP mutant, which was fivefold higher than that of the wild-type virus. This selective advantage was likely to explain its relative stability in polyhedra production for six passages when compared with the wild-type Virus. However, subsequent passaging of ppC19 resulted in a steel) decline in both BV and polyhedra yields, which was also experienced by FP8AS and the wild-type virus Lit high passage numbers. Genomic deoxyribonueleic Licid profiling of the latter suggested that defective interfering particles (DIPS) were implicated in this phenomenon and represented another Undesirable mutation during serial passaging of HaSNPV Hence, a strategy to isolate HaSNPV Clones that exhibited MP-like polyhedra production but FP-like BV production, coupled with low multiplicities of infection during scale-up to avoid accumulation of DIPS, could prove commerically invaluable.

Comparison of apparent ileal amino acid digestibility of feed ingredients measured with broilers, layers, and roosters

The apparent ileal digestibility of amino acids in 7 feed ingredients was determined using broilers, layers, and roosters. The ingredients included 3 cereals (wheat, sorghum, and corn), 3 oilseed meals (canola, cottonseed, and soybean meals), and 1 animal protein meal (meat and bone meal). Dietary protein in the assay diets was supplied solely by the test ingredient. All diets contained 20 g/kg of acid-insoluble ash as an indigestible marker, and each diet was offered ad libitum in mash form to 5 replicate pens of broilers and layers, and 4 replicate pens of roosters. The digestibility coefficients of individual amino acids for wheat, corn, and sorghum were higher (P < 0.05) in broilers than in layers and roosters. The digestibility of most amino acids for corn and sorghum was higher (P < 0.05) in roosters compared with those in layers, whereas the digestibility for wheat in layers and roosters was similar. In general, the digestibility of amino acids in canola meal, cottonseed meal, and meat and bone meal was similar among the 3 classes of chickens. The digestibility of amino acids in soybean meal was higher (P < 0.05) for layers compared with those for broilers and roosters but similar between broilers and roosters. These results suggest that the class of chickens significantly influenced the apparent ileal digestibility of amino acids in some feed ingredients.

Purification and partial characterisation of recombinant human hepcidin

Hepcidin is a liver-expressed antimicrobial and iron regulatory peptide. A number of studies have indicated that hepcidin is important for the correct regulation of body iron homeostasis. The aims of this study were to analyse the expression, trafficking and regulation of human hepcidin in an in vitro cell culture system. Human hepcidin was transfected into human embryonic kidney cells. Immunofluorescence and confocal microscopy analysis revealed that recombinant hepcidin localised to the Golgi complex. Recombinant hepcidin is secreted from the cell within 1 h of its synthesis. Recombinant hepcidin was purified from the cell culture medium using ion-exchange and metal-affinity chromatography and was active in antimicrobial assays. Amino-terminal sequence analysis of the secreted peptide revealed that it was the mature 25 amino acid form of hepcidin. Our results show that recombinant myc-His tagged human hepcidin was expressed, processed and secreted correctly and biologically active in antimicrobial assays. (C) 2005 Elsevier SAS. All rights reserved.