Lethal and sublethal responses of <Emphasis Type="Italic">Sitophilus zeamais</Emphasis> populations to essential oils

The present study aimed to assess the lethal and sublethal responses of five populations of S. zeamais to the essential oils of Ocimum basilicum and Piper hispidinervum, through toxicity, locomotor behavior (flight and walking activities) and physiological (respiration rate and body mass) evaluations. The populations of S. zeamais were obtained from Machado—MG, Paracatu—MG, Piracicaba—SP, Recife—PE and Tunápolis—SC. To estimate the LC₅₀ of each oil for each population, fumigation tests were performed. The population from Recife exhibited the lowest instantaneous rate of increase (ri) as well the lowest consumption of grain mass. However, Piracicaba was considered the susceptibility pattern, presenting the lowest LC₅₀ for both essential oils, no statistical difference was observed among populations or treatments in the flight activity bioassays. The essential oils effect on walking activity and respiration rates varied among populations. The population from Recife presented the highest walked distance and walk speed, with the lowest resting time and number of stops, as well as the lowest respiration rate and body mass. The present study highlights the importance of the study on population responses, as the five populations of S. zeamais exhibited different patterns of toxic and behavioral effects when subjected to the essential oils of P. hispidinervum and O. basilicum.     

Detection of Mycobacterium paratuberculosis antigen with colloidal immunogold in naturally infected sheep.

The anti-Mycobacterium paratuberculosis polyclonal serum is proved useful for labelling Mycobacterium paratuberculosis in glutaraldehyde-osmium-fixed and epon-embedded intestinal samples from sheep with clinical symptoms of paratuberculosis. M. paratuberculosis marked with antibody-coated colloidal gold stain was seen in macrophages, epithelioid cells, giant cells and neutrophils throughout intestinal mucosa. In large macrophages with a low lysosomal content, a great number of intact mycobacteria was seen within phagosomes. In macrophages with average lysosomal content, very few intact mycobacteria or mycobacterial debris were present and lysosome-phagosome fusions were observed. Mycobacteria within neutrophils were scanty. These results show the usefulness of colloidal immunogold techniques for studies of the pathogenesis of paratuberculosis.     

Mapping of quantitative trait loci affecting organ weights and blood variables in a broiler layer cross

1. A genome scan was performed to locate genomic regions associated with traits that are known to vary in birds (most commonly broilers) suffering from heart, lung or muscular dysfunction and for weight of the dressed carcass and some internal organs. 2. The F2 population studied was derived from a cross between a broiler and a layer line and consisted of over 460 birds that were genotyped for 101 markers. 3. There was strong support for segregation of quantitative trait loci (QTL) for carcass and organ weights and blood variables. We identified 11 genome-wide significant QTL (most of them for dressed carcass weight) and several genome-wide suggestive QTL. 4. The results point to some genome regions that may be associated with health-related traits and merit further study, with the final aim of identifying linked genetic markers that could be used in commercial breeding programmes to decrease the incidence of muscular and metabolic disorders in broiler populations.     

Somatic embryogenesis efficiently eliminates viroid infections from grapevines

Indirect somatic embryogenesis is effective at eliminating the most important viruses affecting grapevines. Accordingly, this technique was tested as a method for eradicating two widespread viroids, Grapevine yellow speckle viroid 1 (GYSVd-1) and Hop stunt viroid (HSVd), from four grapevine cultivars. Both viroids were detected by RT-PCR in grapevine floral explants used for initiating embryogenic cultures, as well as in undifferentiated cells of embryogenic and non-embryogenic calli from anthers and ovaries. In contrast, somatic embryos differentiated from these infected calli were viroid-free, and viroids were not detected in embryo-derived plantlets even 3 years after their transfer to greenhouse conditions. A wider spatial distribution of HSVd than GYSVd-1 within proliferating calli was revealed by in situ hybridization, whereas no hybridization signal was detected in the somatic embryos. In addition, GYSVd-1 and HSVd were localised in the nucleus of infected cells, conclusively showing the nuclear accumulation of representative members of Apscaviroid and Hostuviroid genera, which has been only an assumption so far. Somatic embryogenesis was compared to in vitro thermotherapy, a technique routinely used for virus eradication. After thermotherapy, HSVd and GYSVd-1 were detected in all in vitro plantlets of the cultivar Roussan, and in all lines analysed after 3 years of culture in greenhouse. The high efficiency with which somatic embryogenesis may eliminate viroids and viruses from several infected grapevine cultivars, should allow the availability of virus- and viroid-free material, which would be useful not only for sanitary selection but also for basic research on plant-virus and plant-viroid interactions in grapevine.     

Detection of Citrus Leaf Blotch Virus Using Digoxigenin-Labeled cDNA Probes and RT–PCR

Citrus leaf blotch virus (CLBV) was detected by dot-blot hybridization (DBH), and tissue print hybridization (TPH) and by one-step RT–PCR in citrus plants growing both in the greenhouse and in the field. DBH with digoxigenin-labeled cDNA probes allowed CLBV detection in dsRNA-rich and total RNA preparations equivalent to 5 and 0.1mg of infected tissue, respectively. DBH gave intense signals with RNA extracts from young bark, tender shoots and young leaves, whereas the best hybridization signals with TPH were obtained using tender shoots and young leaf petioles. One-step RT–PCR was 10-fold more sensitive than DBH and amplification was obtained with all infected tissues. CLBV was readily detected in young leaves of infected Eureka lemon, Marsh grapefruit, Nules clementine, Navelina orange and Nagami kumquat in the greenhouse, using either hybridization or RT–PCR, but not in leaves of Pineapple sweet orange. Detection in field trees was less consistent and was only achieved by RT–PCR and DBH. CLBV was detected by DBH and RT–PCR in different citrus varieties from several geographic areas showing bud union crease on trifoliate rootstocks, but not in neighbor trees with the same symptoms or in other varieties showing bud union crease on those rootstocks. Failure to detect CLBV in trees with bud union crease could be due to low virus titer or uneven distribution within the plant. Alternatively, a different agent could be involved in causing bud union crease.     

Mating disruption of California red scale,Aonidiella aurantii Maskell (Homoptera: Diaspididae), using biodegradable mesoporous pheromone dispensers

BACKGROUND: The control of California red scale, Aonidiella aurantii (Maskell), has encountered many difficulties, which has raised interest in alternative control methods. Up to now, the A. aurantii sex pheromone has been used only for monitoring. In a previous work the authors described a biodegradable mesoporous pheromone dispenser for mating disruption. To verify the efficacy of these dispensers, three field trials were conducted, and the results are shown in this paper. RESULTS: The study of the release profile of these dispensers revealed a mean pheromone emission value of 269 µg day^?1 and levels of residual pheromone of 10% at the end of 250 days. During the second flight, an A. aurantii male catch reduction of 98% was achieved in the mating disruption plot of trial 1, 93.5% in trial 2 and 76.7% in trial 3. During the third flight, reductions were 94.1, 82.9 and 68.1% in trials 1, 2 and 3 respectively. Considering damaged fruit with more than five scales, reductions of about 80 and 60% were obtained in the mating disruption plots of trials 2 and 3, respectively, compared with an untreated plot, and a reduction of about 70% in trial 1 compared with an oil‐treated plot. CONCLUSION: Mating disruption has been found to be an efficient technique to control this pest, working equally well to a correctly sprayed oil treatment. Further studies are needed to improve the determination of the time of dispenser application and evaluate the effects of the pheromone on natural enemies. Copyright © 2010 Society of Chemical Industry     

Association of citrus psorosis B symptoms with a sequence variant of the Citrus psorosis virus RNA 2

Citrus psorosis virus (CPsV), the type species of genus Ophiovirus, is the presumed causal agent of a bark scaling disease in citrus plants. CPsV virions are kinked filaments composed of three negative‐strand RNA molecules and a ～48‐kDa coat protein. The virus induces two different syndromes: psorosis A (PsA), characterized by limited bark scaling lesions in the trunk and main limbs, and a more aggressive form of the disease called psorosis B (PsB) with rampant bark lesions affecting even thin branches and chlorotic blotches in old leaves. In the greenhouse, the PsA and PsB syndromes can be induced by graft inoculating healthy citrus seedlings with non‐lesion or with lesion bark inoculum from PsA‐affected field trees. PsA‐ and PsB‐inducing CPsV sub‐isolates obtained by this procedure from the same tree showed identical single‐strand conformation polymorphism (SSCP) profiles in homologous segments of the RNAs 1 and 3, whereas segments of the RNA 2 enabled discrimination between PsA‐ and PsB‐associated sequence variants. SSCP analysis of the RNA 2 population present in different tissues of psorosis‐infected plants showed that: (i) PsA‐inducing isolates contain PsB‐associated sequence variants at low frequency, (ii) the PsB‐associated sequence variant is predominant in blistered twigs and gummy pustules affecting old leaves, characteristic of PsB isolates, and (iii) the PsB‐associated sequence variant accumulates preferentially in bark lesions of the trunk and limbs. SSCP analysis of the RNA 2 population also enabled monitoring of interference between PsA‐ and PsB‐associated variants in plants co‐inoculated with both psorosis types.     

From Linderiella baetica to gambilusa: Involving children in conservation by giving a new species a common name

1. The knowledge and awareness that the general public has about aquatic invertebrates is often poor, even when they are highly threatened. The frequent lack of popular names and the unattractiveness of the scientific ones may be among the factors hindering the knowledge of these organisms and, consequently, the awareness of their conservation status and the willingness to conserve them.
2. Linderiella baetica (Crustacea, Branchiopoda, Anostraca) was discovered in 2007 in southern Spain and is considered a critically endangered species. However, being a small invertebrate inhabiting generally unappreciated temporary ponds, this species was completely unknown by the general public, even in the area in which it occurs.
3. To increase the knowledge and awareness of L. baetica, 1,347 children from five schools within its distribution area participated in a contest to select a common name for this newly described taxon. The winning name, gambilusa (Andalusian shrimp), will be used in the future by the Spanish Ministry of Environment and hopefully by the general public to identify the species.
4. The contest was found to have increased the knowledge about the target species in a sustained manner, as 1 year later, 93% of the children recognized gambilusa as a crustacean, 75% remembered its typical habitat, and 89% broadly identified its geographical distribution.
5. This kind of name-choosing contest is applicable for different regions and taxa to raise awareness on the conservation of threatened but poorly known biota.

     

In vitro rumen methane output of perennial ryegrass varieties and perennial grass species harvested throughout the growing season

The selection and feeding of perennial ryegrass (Lolium perenne L.) varieties (PRV) or perennial grass species (PGS) may affect enteric methane (CH₄) output because of changes in the fermentation dynamics in the rumen as a result of differences in herbage chemical composition. The objective of this study was to determine the effects of PRV and PGS harvested throughout the growing season on herbage chemical composition, and in vitro rumen fermentation variables and CH₄ output per unit of feed using a batch culture technique. Seven PRV (Experiment 1: Alto, Arrow, Bealey, Dunluce, Greengold, Malone, Tyrella) and six perennial grasses [Experiment 2: perennial ryegrass (Navan), perennial ryegrass (Portstewart), cocksfoot, meadow fescue, tall fescue, timothy; defined as PGS], managed under a simulated grazing regime, were incubated for 24 h with buffered rumen fluid in two separate experiments. The CH₄ output per unit of feed dry‐matter (DM) incubated was not affected (P > 0·05) by PRV (range of mean values across PRV of 23·9–25·3 (SEM 0·41) mL g^?1 DM) or by PGS (25·6–26·6 (SEM 0·37) mL g^?1 DM). The CH₄ output per unit feed DM disappearing during the in vitro rumen incubation was not affected by PRV (33·9–35·1 (SEM 0·70) mL g^?1 DM), and although there was an overall PGS effect (P < 0·05; 37·2–40·3 (SEM 0·71) mL g^?1 DM), none of the paired contrasts between PGS were significant when analysed using Tukey adjusted comparisons. This outcome reflected either small‐scale or a lack of treatment effects on individual herbage chemical composition (e.g. 454–483 g NDF kg^?1 DM, 215–224 g CP kg^?1 DM and 94–122 g water‐soluble carbohydrate (WSC) kg^?1 DM across PRV; 452–506 g NDF kg^?1 DM, 208–243 g CP kg^?1 DM and 73–131 g WSC kg^?1 DM across PGS) and in vitro rumen fermentation variables. Hence, these results provide no encouragement that choices among the grasses examined, produced within the management regimes operated, would reduce enteric CH₄ output per unit of feed in vivo. However, the technique utilized did not take account of animal × PRV or PGS interactions, such as potential differences in intake between animals, that may occur under farm conditions.