Next-generation sequencing and metagenomic analysis: a universal diagnostic tool in plant virology

A novel, unbiased approach to plant viral disease diagnosis has been developed which requires no a priori knowledge of the host or pathogen. Next-generation sequencing coupled with metagenomic analysis was used to produce large quantities of cDNA sequence in a model system of tomato infected with Pepino mosaic virus . The method was then applied to a sample of Gomphrena globosa infected with an unknown pathogen originally isolated from the flowering plant Liatris spicata . This plant was found to contain a new cucumovirus, for which we suggest the name 'Gayfeather mild mottle virus'. In both cases, the full viral genome was sequenced. This method expedites the entire process of novel virus discovery, identification, viral genome sequencing and, subsequently, the development of more routine assays for new viral pathogens.     

The application of random amplified polymorphic DNA for sandfly species identification

Abstract. We have applied the recently developed Random Amplified Polymorphic DNA (RAPD) method to produce species-specific, DNA profiles for two sympatric, Venezuelan sandfly species, thought to be the vectors responsible for recent outbreaks of cutaneous and mucocutaneous leishmaniasis in the Andean State of Tachira. Moreover, within the profile, it was possible to identify a diagnostic DNA band for Lu.youngi of 0.32 kb. Results showed that the size of this diagnostic DNA band remained constant and did not vary with sex or geographical distribution.     

Does Aversive Conditioning Reduce Human—Black Bear Conflict?

ABSTRACT Aversive conditioning (AC) has the potential to temporarily reduce conflicts between humans and black bears (Ursus americanus). From 2002 to 2005, I evaluated the effectiveness of projectiles with varying impact intensities, pepper spray, and chasing on approximately 150 bears in Sequoia National Park. Aversive conditioning was successful in keeping bears that were not food-conditioned from becoming food-conditioned. For the bears that were already food-conditioned, 17 of 29 bears subjected to AC abandoned unwanted behaviors, 6 required continual treatments, and 6 were killed or relocated. Success with food-conditioned bears was highest when AC was applied soon after bears obtained human food. Aversive conditioning was less successful on yearlings than adults. Rubber slugs were slightly more effective than lower impact projectiles.     

Characterization of body louse midgut proteins recognized by resistant hosts

Abstract. The human body louse, Pediculus humanus , showed eighteen midgut proteins ranging between 12 and 117 kDa, when analysed by SDS-PAGE electrophoresis. Seven of them (12 kDa, 17 kDa, 29 kDa, 35 kDa, 40 kDa, 55 kDa and 97 kDa) were major bands based on their intensity of staining. The immunization of rabbits with a midgut extract elicited the production of protective polyclonal antibodies. These antibodies reacted strongly with all major midgut proteins as well as with 63 kDa and 117 kDa proteins when tested by the Western blot technique. The analysis of the proteins revealed that the 12 kDa, 25 kDa, 29 kDa, 35 kDa, 45 kDa, 87 kDa and 97 kDa proteins are glycosylated and none of them contained a lipid moiety. By electroelution, the proteins of 35 kDa and 63 kDa were purified. On trypsinization, the proteins of 35 kDa and 63 kDa produced four major fragments (F, F₂, F₃, and F₄) when resolved on a 18% SDS-PAGE. The Fj fragment of the 35 kDa protein reacted with me polyclonal antibodies by the immunoblot technique.     

Abundance–range size relationships of macrolepidoptera in Britain: the effects of taxonomy and life history variables

1. A positive relationship between the local abundances and regional distributions of species in an assemblage has been observed for a wide variety of taxa, but its underlying mechanisms remain poorly understood.
2. Taxonomically controlled analyses of a group of the British macrolepidoptera (moths) find a strong, abundance–range size relationship.
3. Degree of polyphagy and habitat generalism explain significant within-taxon variation in both range size and abundance, with polyphages and habitat generalists having larger ranges and greater local abundances than specialists.
4. Taxa that feed in niches considered to be associated with a high diversity of parasitoids have significantly smaller range sizes than other taxa.
5. With the possible exception of body size, none of the life history variables examined explain significant variation around the range size–abundance relationship.     

Effect of plasma components on the feeding response of the mosquito Aedes aegypti L. to adenine nucleotides

ABSTRACT. The gorging response of Aedes aegypti to the ATP dissolved in platelet-poor plasma is greater than that of ATP dissolved in 0.15 m NaCl. The plasma components NaHCO₃ and albumin account for the full effect of the potentiation. Phosphate or tris buffers do not duplicate the bicarbonate effect. In 0.15 m NaCl with bicarbonate but lacking albumin the concentrations inducing 50% feeding are 58 μM ATP, 140 μM ADP, 460 μM AMP and 1500 μM cAMP. Non-adenine nucleotides such as ITP and GTP and phytic acid, and 2,3-diphosphoglyceric acid had no activity.     

Detection of potato virus Y in the aphid Myzus persicae by enzyme-linked immunosorbent assay (ELISA)

Potato virus Y was detected by enzyme linked immunosorbent assay (ELISA) in at least 50% of groups of five Myzus persicae. The mean A₄₀₅ value for groups of viruliferous aphids was 2–3 times greater than that for virus-free ones. PVY was not detected in Aphis craccivora, A. citricola or A. gossypii, three other species which transmitted the virus to peppers, and it was detected in only a small proportion of groups of Acyrthosiphon pisum. In a series of trials, success in detection of PVY by ELISA was not correlated with the ability of other aphids from the same source plant to transmit the virus to test plants. The limitation of ELISA for quantitative assay of PVY in aphids and for epidemiological work is discussed.