No transmission of Potato spindle tuber viroid shown in experiments with thrips (Frankliniella occidentalis, Thrips tabaci), honey bees (Apis mellifera) and bumblebees (Bombus terrestris)

Experiments were carried out to investigate whether Potato spindle tuber viroid (PSTVd) can be transmitted intra- and inter-species from infected Solanum jasminoides to non-infected S. jasminoides and S. esculentum and from infected Brugmansia sp. to S. esculentum by Frankliniella occidentalis and Thrips tabaci by leaf sucking. The F. occidentalis experiments also included feeding on pollen prior to feeding on PSTVd-infected leaf. No thrips-mediated transmission of PSTVd was recorded. The possibility of PSTVd transmission by Apis mellifera and Bombus terrestris during their feeding/pollinating activities within ornamentals and from ornamentals to S. esculentum was included, and no bee-mediated transmission was recorded.     

Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR

Loose smut fungi of barley and wheat (Ustilago nuda and U. tritici, respectively) colonize the plant without causing obvious disease symptoms before heading. The availability of diagnostic methods to detect and follow the growth of these pathogens in the plant would therefore be highly advantageous for both resistance breeding and the development of effective seed treatments. Using seed lots of barley and wheat highly infected with loose smut, we studied the early establishment of the loose smut pathogens in the plant by fluorescence microscopy. In hand-cut sections stained with the fluorochrome Blankophor?, fungal hyphae were observed to invade the shoot apical meristem and leaf primordia during the first days after the onset of germination. At the first node stage the ear and leaf primordia were generally extensively colonized. Hyphae of U. nuda were also regularly observed in high density in the nodes. A protocol was developed for the specific amplification of U. nuda and U. tritici DNA extracted from infected plant tissue. PCR screening of U nuda in seedlings from infected and healthy seed lots was compared to ELISA, microscopy and ultimately head infection of mature plants derived from tillers of the tested seedlings. The results indicated that a prediction of loose smut infection by real-time PCR is possible at the second leaf stage, and that the assay is equally suited for use with spring and winter varieties of barley and wheat.     

Structural and physiological changes associated with the skin spot disorder in apple

Skin spot is an important physiological disorder of ‘Elstar’ apples (Malus × domestica Borkh.) that occurs after fruit have been removed from controlled atmosphere storage. Skin spots are irregular patches of small, round, brown blemishes. Cross-sections reveal a browning of protoplasts (coagulated) and of cell walls that extends into the hypodermis. Skin spots are always associated with linear, gaping and non-gaping microcracks in the cuticle. Staining of apple skin with calcofluor white usually results in white fluorescence of cell walls but, within a skin spot, the white fluorescence is weak or absent. Cell walls within, and in the immediate vicinity of skin spots stain with phloroglucin/HCl indicating the presence of lignin. The area of skin affected by skin spots was positively and linearly correlated with the area of the non-blush fruit surface infiltrated by acridine orange. In general, skin spots were limited to the non-blush fruit surface and occurred more frequently near the stem-end than the calyx region of the fruit. Skin spot areas were correlated with a 2.5-fold increase in water vapour permeability compared with unaffected areas (23.8 ± 4.0 m s⁻¹ with skin spots, 9.6 ± 2.1 × 10⁻⁵ m s⁻¹ without skin spots). Strips of the fruit skin from regions with skin spots had an increased maximum force and modulus of elasticity. Dipping fruit in ascorbic acid (0.1 or 0.3 mM for 10 min) before storage decreased the area affected by skin spots. There was no effect of dipping in ethanol/water (70%, v/v, 15 min) or in solutions of captan (1.5 g L⁻¹, 10 min) or trifloxystrobin (0.1 g L⁻¹, 10 min). In contrast, prestorage treatment with 1-methylcyclopropene (630 nL L⁻¹ for 24 h) or poststorage incubation in H₂O₂ (10% for 2, 6, 10 and 16 h) increased skin spots. Our data are consistent with a typical cell response to an oxidative burst that seems to be focussed on particular regions of the ‘Elstar’ fruit surface by concentrations of cuticular microcracks, and that is possibly caused by reoxygenation injury upon removal from CA storage.     

Cavity cooling below the recoil limit

Conventional laser cooling relies on repeated electronic excitations by near-resonant light, which constrains its area of application to a selected number of atomic species prepared at moderate particle densities. Optical cavities with sufficiently large Purcell factors allow for laser cooling schemes, avoiding these limitations. Here, we report on an atom-cavity system, combining a Purcell factor above 40 with a cavity bandwidth below the recoil frequency associated with the kinetic energy transfer in a single photon scattering event. This lets us access a yet-unexplored regime of atom-cavity interactions, in which the atomic motion can be manipulated by targeted dissipation with sub-recoil resolution. We demonstrate cavity-induced heating of a Bose-Einstein condensate and subsequent cooling at particle densities and temperatures incompatible with conventional laser cooling.     

Asymmetric segregation of polarized antigen on B cell division shapes presentation capacity

During the activation of humoral immune responses, B cells acquire antigen for subsequent presentation to cognate T cells. Here we show that after mouse B cells accumulate antigen, it is maintained in a polarized distribution for extended periods in vivo. Using high-throughput imaging flow cytometry, we observed that this polarization is preserved during B cell division, promoting asymmetric antigen segregation among progeny. Antigen inheritance correlates with the ability of progeny to activate T cells: Daughter cells receiving larger antigen stores exhibit a prolonged capacity to present antigen, which renders them more effective in competing for T cell help. The generation of progeny with differential capacities for antigen presentation may have implications for somatic hypermutation and class switching during affinity maturation and as B cells commit to effector cell fates.     

An unusual outbreak of histomonosis in a commercial turkey flock

In the past histomonosis was very well controlled with Dimetridazole as a treatment and/or Nifursol as feed additive. In the European Union both products were banned in 1995 and 2003, respectively. This was followed by the re-emergence of the disease in the recent years. In the present case a farm with two houses was affected by the disease. In each house 2620 hens and 2620 toms were kept, separated by wire mesh. At the 53rd day of age the toms in house 1 showed general clinical symptoms, accompanied by a slightly raised mortality, which sharply increased in the following days. At necropsy all dead birds showed lesions typical for histomonosis in caeca and liver. Histomonosis was diagnosed by histopathology and PCR. Within five days cumulative mortality was 25.1%.The hens kept at the same house didn't show any symptoms. At day 57 two toms, which were kept in house 2, died and showed similar symptoms and lesions. Within the next three days 48 more birds died. Again the hens in house 2 showed neither clinical signs nor mortality. Treatment trials using herbal products and a change of litter directly after the onset of clinical signs did not reduce the mortality. On day 62 the toms of both houses were euthanized by CO2 in closed containers. The hens were kept until they were slaughtered in week 16 and did not show any evidence of histomonosis.     

Comparative spatiotemporal analysis of the intrathecal immune response in natural listeric rhombencephalitis of cattle and small ruminants

This study examined the spatiotemporal immune response in listeric rhombencephalitis of ruminants in situ. Our data support the view that astrocytes facilitate the containment of infectious lesions. Results on the natural disease recapitulate observations in experimental rodent models and suggest that the mounted adaptive lymphocytic response of ruminants is effective in eliminating Listeria monocytogenes (LM). However, our data indicate earlier participation of the adaptive immune response, a stronger B lymphocyte contribution and a more protracted macrophage infiltration in the natural disease than it has been deduced from experimental models. Therefore, such models should be complemented by studies in natural host systems. Various macrophage and microglia subsets are involved in listeric rhombencephalitis and their differential contribution may account for species differences in clinical course and outcome of infection as might species differences in the B-cell response. Future functional ex vivo and in vitro studies are necessary to further investigate the findings obtained in the present study.