Detection of hepatitis E virus shedding in feces of pigs at different stages of production using reverse transcription-polymerase chain reaction.

The aim of this study was to determine at which production stages hepatitis E virus (HEV) is shed by the highest number of pigs and to estimate the relative risk associated with each stage. For this purpose, 146 fecal samples of pigs from 21 farms were studied. In addition, 1 sample from the manure ditch and another sample of drinking water, collected directly from the trough located in the pen, were taken from 16 farms. HEV RNA was detected in fecal samples from 34 pigs (23.29%). The production stages in which most pigs excreted HEV were weaners (41.7%) and pigs in the first month of feeding (60%). The results of the statistical analysis showed that the principal significant risk stage in HEV shedding was the first month of feeding (odds ratio [OR] 19.5, 95% CI 3.59-106.07, P = 0.001) followed by the weaners stage (OR 9.3, 95% CI .78-48.42, P = 0.008). In 8 out of 16 farms tested (50%) HEV RNA was detected in raw manure and in the water trough of only 1. Detection of HEV in manure ditches raises the concern of how to deal with manure of swine origin, because it is used as soil fertilizer.     

Comparison ofPotato Virus Y andPlum Pox Virus transmission by two aphid species in relation to their probing behavior

Two different aphid species,Myzus persicae (Sulzer) andHyalopterus pruni (Geoffroy) (Homoptera: Aphididae), were used to analyze their ability to transmit two different potyviruses,Potato virus Y (PVY) andPlum pox virus (PPV), to pepper (Capsicum annuum) andNicotiana benthamiana plants, respectively. In parallel experiments,M. persicae consistently transmitted both viruses with high efficiency, whereasH. pruni always failed to transmit either virus. This is in contrast to previous reports describingH. pruni as a vector of these potyviruses. Different aphid probing behavior among individual aphids of each species was obtained in electrical penetration graph (EPG) experiments performed on pepper plants. This suggested thatH. pruni did not transmit these potyviruses due to behavioral differences during probing that impeded virus acquisition and/or inoculation. It was found thatM. persicae usually makes its first probe within the first 2 min, whereasH. pruni individuals remained for more than 10 min on the plant before starting to probe. Furthermore,M. persicae individuals displayed their first intracellular puncture during the first minute of probing whereasH. pruni needed ∼ 15 min to penetrate the cell plasmalemma with their stylets. In addition, intracellular stylet punctures occurred very frequently forM. persicae but was a rare event, never exceeding a single one, forH. pruni. The relevance of these findings for the epidemiological spread of potyviruses by different aphid species is discussed. http://www.phytoparasitica.org posting May 14, 2006.     

Genetic variation of Prunus avium in susceptibility to cherry leaf spot (Blumeriella jaapii) in spatially heterogeneous infected seed orchards

Cherry leaf spot (Blumeriella jaapii (Rehm) Arx.) is a very serious disease of wild cherry (Prunus avium L.), which produces premature leaf defoliation and vigor decrease. In two clonal seed orchards of P. avium naturally infected by B. jaapii, spatial heterogeneity and autocorrelation of neighbor damage caused by cherry leaf spot impeded proper analysis of the fungus incidence. The iterative spatial analysis (ISA), based on variography and kriging, was successfully used to eliminate the effect of this spatial heterogeneity in analysis of genetic variation in susceptibility to B. jaapii. Significant differences among P. avium clones were found, with moderate to high broad-sense heritability estimates. Genetic by environment interactions, although significant, were not quantitatively important. A strong relationship between leaf spot susceptibility and bud burst was found. However, other factors must be affecting the genetic variation in leaf spot susceptibility, as differences among clones remained highly significant when considering the bud burst as a covariate in the genetic model.     

Effects of nitrogen fertilization and temperature on frost hardiness of Aleppo pine (Pinus halepensis Mill.) seedlings assessed by chlorophyll fluorescence

In a 14-week study, 1-year-old Aleppo pine seedlings were grownin two growth chambers. Seedlings were artificially hardenedby decreasing photoperiod and temperature. In each chamber halfof the seedlings were fertilized with nitrogen (8.4 mg seedling^–1).In order to determine the relative importance of the hardeningenvironment versus fertilization, each chamber was programmedto decrease night temperatures down to a low of 8 or 4°C.Chlorophyll fluorescence and frost hardiness was measured fivetimes during the experiment. A sample of seedlings from eachtreatment was exposed to an artificial frost at –5°Cand the freezing effects were assessed by measurements of chlorophyllfluorescence and visual evaluation of needle damage. Seedlingsincreased their frost hardiness during the experiment in allthe treatments but the ratio of variable to maximal chlorophyllfluorescence (F_v/F_m) measured before freezing did not vary duringthe experiment. This indicates that Aleppo pine maintains itsphotosynthetic ability during hardening in contrast to otherconiferous species from colder climates. The effect of nitrogenfertilization on frost hardiness was small in comparison withchamber effect. Nitrogen fertilization slightly delayed theacquisition of hardening in the coldest chamber. Seedlings inthe warmest chamber did not become fully resistant to –5°C,but in the coldest chamber, where night temperature reached4°C, all the seedlings were resistant to the frost. Severedamage caused by frost could be related to a rapid rise of minimalfluorescence (F₀) but the best index of damage was the dropof F_v/F_m after freezing.     

Changes in understory species diversity during theNothofagus pumilio forest management cycle

Human commercial activities must be compatible with sustainable management to achieve species conservation in the natural environment. After harvesting, diversity and abundance of understory species differ substantially from the pre-harvest situation. Have knowledge of the changes that logging produces in all components of the system allows us to develop management alternatives with reduced impacts. The objective of this work was to evaluate changes in density, biomass and diversity of understory species along a forest shelterwood cut system management cycle in aNothofagus pumilio forest. Six forest management situations (postharvesting stands and different phases of forest development) were sampled in a pureN. pumilio forest of Tierra del Fuego (Argentina) of middle-high site class (fromclass I toIII). Species number, density and biomass were modified substantially after a shelterwood cut. These were being greatest after cutting and falling substantially when the forest canopy closes during theinitial growth phase, returning to values similar to this virgin forest when themature phase was reached. The impact of the forest management on understory species diversity is analyzed and discussed. The silvicultural practices applied toN. pumilio forest in Tierra del Fuego permit the introduction of alien flora to the system, and produce great changes during the forest management cycle. To Celina Matteri, Itatí Ferraro, and Rosa Kofalt for their aid in taxonomical identification. To Juan Manuel Cellini, Ricardo Vukasovic and Gregorio Martínez Pastur for their participation in the field work; and to the sawmillLos Castores, Universidad Nacional de la Patagonia Austral, Centro Austral de Investigaciones Científicas and Instituto Nacional de Tecnología Agropecuaria for the institutional and financial support.     

Organogenic responses of Pinus pinea cotyledons to hormonal treatments: BA metabolism and cytokinin content

Isolated cotyledons from mature Pinus pinea L. embryos were cultured in vitro in a factorial combination of 4.4, 10 and 44.4 microM N6-benzyladenine (BA) for 2, 4, 8, 16 and 35 days to optimize shoot regeneration. Incubation of explants in 44.4 microM BA for 4 days, in place of the standard incubation in 4.4 microM BA for 35 days, reduced the entire culture period to 4 weeks. Shortening the culture period had no significant effect on the caulogenic response or the number of buds formed per cotyledon. To establish the relationship between key moments in the caulogenic process induced by 4.4 microM BA and the endogenous concentrations of the active forms of BA and other isoprenoid-type cytokinins (CKs), we examined uptake, metabolism and amount of BA, as well as the amounts of zeatin, dihydrozeatin and their ribosides in P. pinea cotyledons after 1, 2, 6, 12 and 24 h, and 2, 4, 8, 16 and 35 days of exposure to 8-[14C]BA. Uptake and release of BA were associated with water movement between explants and the medium during the first 8 days of culture. The interconvertible forms of BA were the main metabolites formed in the tissues. Inactivation of BA as a result of conjugation or oxidation was insignificant. The endogenous concentration of BA + N6-benzyladenosine was 20-fold higher than the exogenously applied BA during the competence acquisition phase (Days 0-3). The concentration of isoprenoid-type CKs also increased 16-fold and then decreased during this time. Induction of shoot buds (Days 4-8) was characterized by a second peak of BA uptake by explants that triggered the synthesis of N6-benzyladenosine-5 -monophosphate and by the maintenance of isoprenoid-type CKs. Reestablishment of CK homeostasis marked the shift from the induction phase to the shoot development phase in this organogenic process (Days 8-12).     

Effects of various densities of Ophiostoma ips inoculations on Pinus sylvestris in north‐western Spain

The aim was to determine the inoculation density above which Scots pine (Pinus sylvestris) is overcome by the blue‐stain fungus Ophiostoma ips that is associated with the bark beetle Ips sexdentatus. In north‐western Spain, stems of 16 Scots pines were inoculated at various densities (0, 400, 800 or 1600 inoculi/m²) along circumferential 100 or 150 cm wide inoculation belts. Each inoculum consisted of a 5 mm diameter cylinder of malt extract agar colonized by the fungus. Three months later, all trees were harvested and trunk resinosis and foliage colour were visually assessed. The percentage of healthy, desiccated, resin soaked, and blue‐stained sapwood, as well as growth productivity indices, were calculated from stem disks cut from within the inoculated zone of each tree. Sapwood‐specific hydraulic conductivity (Ks) of each tree was measured in the middle of the inoculated zone. All parameters of tree vigour changed dramatically to the worse when inoculation densities were above 400 inoculi/m², and foliage changed from green to yellow‐green or yellow when an inoculation density of 800 instead of 400 was used. The percentage loss of sapwood‐specific conductivity (PLC) increased from 30 to 90% and the percentage of healthy, conductive sapwood dropped from 85 to 35% at 800 inoculi/m². No effect of the width of the inoculation belt was observed, and there was no relationship between tree productivity indices and the level of resistance. A non‐linear negative relationship was found between PLC and the percentage of healthy sapwood. It is concluded that tree resistance was overcome and that trees were going to die when the inoculation density was ≥800 inoculi/m².