Hairpin loop structure of African swine fever virus DNA.

The ends of African swine fever virus genome are formed by a 37 nucleotide-long hairpin loop composed, almost entirely, of incompletely paired A and T residues. The loops at each DNA end were present in two equimolar forms that, when compared in opposite polarities, were inverted and complementary (flip-flop), as in the case of poxvirus DNA. The hairpin loops of African swine fever and vaccinia virus DNAs had no homology, but both DNAs had a 16 nucleotide-long sequence, close to the hairpin loops, with an homology of about 80%. An analysis of African swine fever virus replicating DNA showed head-to-head and tail-to-tail linked molecules that may be replicative intermediates.     

Assembly of Bacillus subtilis phage phi29. 1. Mutants in the cistrons coding for the structural proteins.

The effect of mutations in the cistrons coding for the phage structural proteins has been studied by analyzing the phage-related structures accumulated after restrictive infection. Infection with susmutants in cistron 8, lacking both the major head and the fiber protein, does not produce any phage-related structure, suggesting a single route for the assembly of phage phi29; infection with ts mutants in this cistron produces isometric particles. Mutants is cistron 9, coding for the tail protein, TP1, produce DNA-free prolate heads with an internal core; these particles are abortive and contain the head proteins HPO, HP1 and HP3, the upper collar protein NP2 and the nonstructural proteins p7, p15 and p16. Mutants in cistron 10, coding for the upper collar protein, NP2, produce DNA-free isometric heads also with an internal core; they contain the head proteins and the nonstructural protein p7, suggesting that this protein forms the internal core. Mutants in cistrons 11 and 12, coding for the lower collar protein, NP3, and the neck appendages, NP1, respectively, give rise to the formation of DNA-containing normal capsids and DNA-free prolate particles, more rounded at the corners than the normal capsids and with an internal core; the DNA-containing 11-particles are formed by the head proteins and the upper collar protein; the DNA-free 11-particles contain, besides these proteins, the nonstructural protein p7 and a small amount of proteins p15 and 16. The DNA-containing 12-particles have all the normal phage structural proteins except the neck appendages, formed by protein NP1; the DNA-free particles are similar to the DNA-free 11-particles. After restricitive infection mutant sus14(1241) has a delayed lysis phenotype and produces a phage burst higher than normal, after artificial lysis. It produces DNA-containing particles, identical to wild-type phage, which have all the normal phage structural proteins, and DNA-free prolate particles, more rounded at the corners than the final phage particles and with an internal core; the last particles contain the same proteins as the DNA-free 11 or 12-particles. These particles could represent a prohead state, ready for DNA encapsulation. None of the DNA-containing particles have the nonstructural proteins p7, p15 or p16, suggesting that these proteins are released from the proheads upon DNA encapsulation.     

Toxicity of Fipronil to the Predatory Lacewing Chrysoperla carnea (Neuroptera: Chrysopidae)

All the developmental stages of Chrysoperla carnea (Stephens) were treated with fipronil using different modes of exposure under laboratory conditions. Eggs were dipped in an aqueous range of concentrations and no effects were recorded, except at the highest concentration. Pupae treated topically on the silk cocoon moulted to healthy adults, without any deleterious effects on their reproduction. In contrast, larvae and adults were killed by the compound, irrespective of the mode of treatment, even at rates below the maximum field recommended rate in Spain (30 g c.p./ha). Sublethal concentrations of fipronil did not affect the fecundity or fertility of survivors. We conclude that fipronil is very toxic under laboratory conditions to this predatory lacewing.     

The African swine fever virus prenyltransferase is an integral membrane trans-geranylgeranyl-diphosphate synthase.

In a previous study, it was shown that the protein encoded by the gene B318L of African swine fever virus (ASFV) is a trans-prenyltransferase that catalyzes in vitro the condensation of farnesyl diphosphate and isopentenyl diphosphate to synthesize geranylgeranyl diphosphate and longer chain prenyl diphosphates (Alejo, A., Yá?ez, R. J., Rodríguez, J. M., Vi?uela, E., and Salas, M. L. (1997) J. Biol. Chem. 272, 9417-9423). To investigate the in vivo function of the viral enzyme, we have determined, in this work, its subcellular localization and activity in cell extracts. Two systems were used in these studies: cells infected with ASFV and cells infected with a recombinant pseudo-Sindbis virus carrying the complete B318L gene. In this latter system, the trans-prenyltransferase was found to colocalize with the endoplasmic reticulum marker protein-disulfide isomerase, whereas in cells infected with ASFV, the viral enzyme was present in cytoplasmic viral assembly sites, associated with precursor viral membranes derived from the endoplasmic reticulum. In addition, after subcellular fractionation, the viral enzyme partitioned into the membrane fraction. Extraction of membrane proteins with alkaline carbonate and Triton X-114 indicated that the ASFV enzyme behaved as an integral membrane protein. The membrane enzyme synthesized predominantly all-trans-geranylgeranyl diphosphate from farnesyl diphosphate and isopentenyl diphosphate. These results indicate that the viral B318L protein is a trans-geranylgeranyl-diphosphate synthase, being the only enzyme of this type that is known to have a membrane localization.     

Comparison of the A-T rich regions and the Bacillus subtilis RNA polymerase binding sites in phage phi 29 DNA.

By using a modification of the BAC spreading method for mounting the DNA for electron microscopy, partial denaturation maps of protein-free phi 29 DNA and of phi 29 DNA containing protein p3 were obtained. In phi 29 p3-DNA1 the protein does not seem to influence the melting of the ends of the molecules. The comparison of the partial denaturation map and the B. subtilis RNA polymerase binding sites indicates that five of the seven early promoters (A1, A2, A3, B2 and C2) are located in A-T rich DNA regions whereas the other two early promoters (B1 and C1) are located in less A-T rich sites.     

Effect of Game Management on Wild Red-Legged Partridge Abundance

The reduction of game and fish populations has increased investment in management practices. Hunting and fishing managers use several tools to maximize harvest. Managers need to know the impact their management has on wild populations. This issue is especially important to improve management efficacy and biodiversity conservation. We used questionnaires and field bird surveys in 48 hunting estates to assess whether red-legged partridge Alectoris rufa young/adult ratio and summer abundance were related to the intensity of management (provision of supplementary food and water, predator control and releases of farm-bred partridges), harvest intensity or habitat in Central Spain. We hypothesized that partridge abundance would be higher where management practices were applied more intensively. Variation in young/adult ratio among estates was best explained by habitat, year and some management practices. Density of feeders and water points had a positive relationship with this ratio, while the density of partridges released and magpies controlled were negatively related to it. The variables with greatest relative importance were feeders, releases and year. Variations in post-breeding red-legged partridge abundance among estates were best explained by habitat, year, the same management variables that influenced young/adult ratio, and harvest intensity. Harvest intensity was negatively related to partridge abundance. The other management variables had the same type of relationship with abundance as with young/adult ratio, except magpie control. Variables with greatest relative importance were habitat, feeders, water points, releases and harvest intensity. Our study suggests that management had an overall important effect on post-breeding partridge abundance. However, this effect varied among tools, as some had the desired effect (increase in partridge abundance), whereas others did not or even had a negative relationship (such as release of farm-reared birds) and can be thus considered inefficient or even detrimental. We advise reconsidering their use from both ecological and economical points of view.     

Toxicity and pharmacokinetics of insect growth regulators and other novel insecticides on pupae of Hyposoter didmator (Hymenoptera: Ichneumonidae), a parasitoid of early larval instars of lepidopteran pests

Susceptibility of the lepidopteran parasitoid Hyposoter didymator (Thunberg) to seven modern insecticides, azadirachtin, diflubenzuron, halofenozide, methoxyfenozide, pyriproxyfen, tebufenozide, and spinosad, was tested in the laboratory. Pupae were exposed to different doses of each compound by direct topical application. At the field recommended doses, methoxyfenozide and tebufenozide had no effect on H. didymator. Halofenozide had a low effect on both adult emergence and adult survival but the progeny size and parasitism capacity were not affected. Diflubenzuron was moderately toxic to the parasitoid, while azadirachtin, pyriproxyfen and spinosad were very toxic, affecting all its life parameters. In the pyriproxyfen and spinosad treatments, no progeny was obtained. As a second approach of this study, we determined the rate of penetration through the pupal cocoon and absorption in the parasitoid body as pharmacokinetic parameters important for toxicity. Most of the radioactivity was retained in the silken cocoon, indicating a low accumulation in the parasitoid body. Among all compounds tested, diflubenzuron exhibited the highest absorption in the parasitoid body, followed by pyriproxyfen. For halofenozide, methoxyfenozide and tebufenozide, low absorption (<2%) was found. In addition, we tested for the presence of molting hormone receptors in Hyposoter tissues using a monoclonal antibody 9B9. Our data suggest that the use of diflubenzuron azadirachtin, pyriproxyfen, halofenozide, and spinosad in combination with H. didymator in integrated pest management (IPM) programs should be carefully evaluated. Methoxyfenozide and tebufenozide could be considered safe for this parasitoid.     

The Quest for a Suitable Host: Size Distributions of Host Trees and Secondary Hemiepiphytes Search Strategy

As climbing plants lack the capacity to hold themselves upright, they need to encounter a suitable host. Vines, lianas, and secondary hemiepiphytes need, therefore, an effective searching strategy. Various hypotheses have been put forward on searching strategies, including ‘skototropism’—growth toward darkness—and random searching. We studied host searching strategies of three secondary hemiepiphyte species belonging to the genus Heteropsis. We recorded information on the diameter distribution of host and evaluated whether these hosts were ‘suitable’, i.e., sufficiently tall for Heteropsis individuals to reach reproductive size. The diameter distribution of host trees bearing Heteropsis seedlings was similar to that of the trees in our study plots. Also, we found that 72–81 percent of the Heteropsis seedlings were present on unsuitable hosts (seedlings, saplings, herbs). These results suggest that Heteropsis seedlings search hosts in a random manner and not by skototropism. We found quite a distinct pattern for adult Heteropsis individuals, which predominantly occur on host trees bigger than 10 cm dbh. Host diameter distribution for Heteropsis adults differed significantly from that of the entire tree community. This difference suggests that Heteropsis individuals may change hosts if they are initially present on nonsuitable hosts. We observed that Heteropsis seedlings and juveniles on unsuitable hosts often produced vegetative shoots that searched for another host. In many cases, such shoots did not find a suitable host. For Heteropsis, our results suggest that host tree searching is a long‐term trial and error process that is governed by a random searching strategy. Abstract in Spanish is available in the online version of this article.     

Optimization of the enzymatic synthesis of structured triacylglycerols rich in docosahexaenoic acid at sn-2 position by acidolysis of Aurantiochytrium limacinum SR21 oil and caprylic acid using response surface methodology

Journal of Applied Phycology - The thraustochytrid Aurantiochytrium limacinum SR21 is a promising source of docosahexaenoic acid (DHA) for human consumption as dietary supplement, replacing...