Replacements of Pro86 in phage T4 lysozyme extend an alpha-helix but do not alter protein stability

To investigate the relation between protein stability and the predicted stabilities of individual secondary structural elements, residue Pro86 in an alpha-helix in phage T4 lysozyme was replaced by ten different amino acids. The x-ray crystal structures of seven of the mutant lysozymes were determined at high resolution. In each case, replacement of the proline resulted in the formation of an extended alpha-helix. This involves a large conformational change in residues 81 to 83 and smaller shifts that extend 20 angstroms across the protein surface. Unexpectedly, all ten amino acid substitutions marginally reduce protein thermostability. This insensitivity of stability to the amino acid at position 86 is not simply explained by statistical and thermodynamic criteria for helical propensity. The observed conformational changes illustrate a general mechanism by which proteins can tolerate mutations.     

Effect of dietary echium oil on growth, fatty acid composition and metabolism, gill prostaglandin production and macrophage activity in Atlantic cod (Gadus morhua L.)

Echium oil (EO) is a vegetable oil in which percentages of stearidonic acid (STA, 18:4n‐3) often exceed those of its n‐6 series equivalent γ‐linolenic acid (GLA, 18:3n‐6). Stearidonic acid is elongated to 20:4n‐3 in fish cell cultures, suggesting that EO could be included in diets for marine fish to increase tissue 20:4n‐3 and 20:3n‐6 and, thereby, modulate eicosanoid metabolism. Thus, the present study aimed to test the hypotheses that dietary EO would increase tissue 20:4n‐3 and 20:3n‐6 and modulate immune function and eicosanoid production in juvenile Atlantic cod (Gadus morhua L.) fed a diet where fish oil (FO) was replaced by EO. Duplicate groups of juvenile cod (initial weight ca. 4 g) were fed for 18 weeks on fish meal‐based diets (55% protein and 16% lipid) that differed in oil source (FO or EO). There were no significant differences in growth and feed efficiency, hepato‐somatic index, percentages of liver and flesh lipids and lipid class compositions for cod fed FO and EO. Percentages of 18:4n‐3, 18:3n‐6 and 20:3n‐6 in the total lipids of flesh and liver were higher, and percentages of 20:5n‐3 and 20:4n‐6 were both lower in fish fed EO than in those given FO. In flesh, the increased 18:3n‐6 and 18:4n‐3 were primarily located in phosphatidylcholine and, to a lesser extent, phosphatidylethanolamine, whereas 20:3n‐6 concentration was highest in phosphatidylinositol. Desaturation of 18:3n‐3 (to tetraene products) and 20:5n‐3 to 22:6n‐3 in hepatocytes was very low but was increased by dietary EO. Echium oil significantly decreased the production of prostaglandin F from gill cells stimulated with calcium ionophore A23187, and reduced head kidney macrophage activity, but had no effect on serum lysozyme activity or basic haematology. In conclusion, dietary EO may have beneficial effects on some immune parameters including eicosanoid metabolism in marine fish although this may be primarily because of decreased 20:4n‐6 rather than increasing tissue levels of 20:3n‐6 or 20:4n‐3.     

Scent organ development in creatonotos moths: regulation by pyrrolizidine alkaloids

7-Hydroxy-6,7-dihydro-5H-pyrolizine-1-carboxaldehyde is the major volatile component of the scent organs in males of two species of Creatonotos (Lepidoptera, Arctiidae). The biosynthesis of this presumed pheromone depends on the presence of pyrrolizidine alkaloids in plants that are ingested by the larvae. In addition, these secondary plant substances control the morphogenesis of the scent organs. This morphogenetic effect of an alkaloid has not been observed previously.     

Biological Control of Yellow Nutsedge with the Indigenous Rust Fungus Puccinia canaliculata

Yellow nutsedge (Cyperus esculentus L.) is a serious weed problem in the United States and other countries. An indigenous rust fungus [Puccinia canaliculata (Schw.) Lagerh.], pathogenic on yellow nutsedge, was released in early spring as a potential biological control agent. The fungus inhibited nutsedge flowering and new tuber formation. The fungus also dehydrated and killed nutsedge plants. The successful control of yellow nutsedge by a rust epiphytotic under experimental conditions demonstrates the potential use of the rust in an integrated weed management system.     

Australian surveillance for avian influenza viruses in wild birds between July 2005 and June 2007

Objective   To identify and gain an understanding of the influenza viruses circulating in wild birds in Australia.
Design   A total of 16,303 swabs and 3782 blood samples were collected and analysed for avian influenza (AI) viruses from 16,420 wild birds in Australia between July 2005 and June 2007. Anseriformes and Charadriiformes were primarily targeted.
Procedures   Cloacal, oropharyngeal and faecal (environmental) swabs were tested using polymerase chain reaction (PCR) for the AI type A matrix gene. Positive samples underwent virus culture and subtyping. Serum samples were analysed using a blocking enzyme-linked immunosorbent assay for influenza A virus nucleoprotein.
Results   No highly pathogenic AI viruses were identified. However, 164 PCR tests were positive for the AI type A matrix gene, 46 of which were identified to subtype. A total of five viruses were isolated, three of which had a corresponding positive PCR and subtype identification (H3N8, H4N6, H7N6). Low pathogenic AI H5 and/or H7 was present in wild birds in New South Wales, Tasmania, Victoria and Western Australia. Antibodies to influenza A were also detected in 15.0% of the birds sampled.
Conclusions   Although low pathogenic AI virus subtypes are currently circulating in Australia, their prevalence is low (1.0% positive PCR). Surveillance activities for AI in wild birds should be continued to provide further epidemiological information about circulating viruses and to identify any changes in subtype prevalence.     

The ecoresponsive genome of Daphnia pulex

We describe the draft genome of the microcrustacean Daphnia pulex, which is only 200 megabases and contains at least 30,907 genes. The high gene count is a consequence of an elevated rate of gene duplication resulting in tandem gene clusters. More than a third of Daphnia's genes have no detectable homologs in any other available proteome, and the most amplified gene families are specific to the Daphnia lineage. The coexpansion of gene families interacting within metabolic pathways suggests that the maintenance of duplicated genes is not random, and the analysis of gene expression under different environmental conditions reveals that numerous paralogs acquire divergent expression patterns soon after duplication. Daphnia-specific genes, including many additional loci within sequenced regions that are otherwise devoid of annotations, are the most responsive genes to ecological challenges.     

Flaxleaf fleabane (Conyza bonariensis) populations have developed glyphosate resistance in north-east Australian cropping fields

In recent years flaxleaf fleabane has become a widespread and difficult-to-control weed in no-tilled fallowed fields, where weeds are controlled by applications of glyphosate, in annual cropping systems of north-east Australia. Fifty-two populations, collected in a national survey from agricultural and non-agricultural areas, were tested in two glyphosate dose-response pot experiments. In two subsequent pot experiments, a sub-set of these populations was tested with a field rate of glyphosate when weeds of two ages were grown at different soil moistures. In the first and second experiments, most populations collected from chemical fallowed or cropped fields in north-east Australia had GR₅₀ (estimated dose for 50% biomass reduction) values three to six times greater than the susceptible populations, indicating low levels of glyphosate resistance. Several populations from roadsides adjacent to chemical fallowed or cropped fields also had higher GR₅₀ values, indicating movement of seeds from resistant plants. In the third experiment, weed biomass of all populations from chemical fallowed or cropped fields was 70-98% of unsprayed compared to 2-3% for the susceptible populations, irrespective of weed age or soil moisture. In the fourth experiment which treated older weeds, the response of several resistant populations to glyphosate was unaffected by differences in weed age and soil moisture, whereas the biomass of the other resistant populations was greater following spraying of older and/or moisture stressed plants compared with smaller non-stressed plants. Thus, exclusive reliance on glyphosate for fallow weed control in this region has resulted in the evolution of resistance in flaxleaf fleabane populations in a cropping system with annual non-transgenic crops. Prolific production of windborne seeds, combined with poor control associated with spraying large moisture-stressed weeds, is likely to have contributed also to flaxleaf fleabane becoming such a problem weed.     

Inactivation at various temperatures of bovine viral diarrhea virus in beef derived from persistently infected cattle

Bovine viral diarrhea virus (BVDV) is a pestivirus that is enzootic in most cattle populations throughout the world. This virus is present throughout the body of persistently infected (PI) cattle. Previous research has not assessed the cooking temperature at which BVDV in meat from PI cattle can be inactivated. Therefore, muscle tissue from 6 PI cattle was harvested, refrigerated, frozen, and heated to various internal temperatures. The concentration of virus present was determined by virus isolation. Average cell culture infective doses (50% endpoint; CCID(50)) of BVDV per gram of frozen, uncooked meat from PI cattle were 10(5.85) CCID(50)/g of whole cuts and 10(6.02) CCID(50)/g of ground meat. The virus in whole and ground meat was consistently inactivated when cooked to temperatures greater than or equal to 75°C. A second objective of this research was to thoroughly reassess if Vero cells were permissive to BVDV infection in our laboratory to provide further indication of whether primates, including humans, might be susceptible to BVDV. Vero cells were not permissive to infection with any of 43 different strains of BVDV that readily replicated in Madin Darby bovine kidney cells. In conclusion, this bovine pathogen, which is not considered to be a human pathogen, can be inactivated by cooking ground or whole cuts of meat to 75°C or higher. Care should be taken to ensure that susceptible hosts such as pigs are not fed improperly cooked meat, meat by-products, or waste food originating from PI cattle.