Structure and temporal shifts in virulence of Pseudoperonospora cubensis populations in the Czech Republic

The structure and temporal dynamics of the virulence of Pseudoperonospora cubensis (causal agent of cucurbit downy mildew) were studied in pathogen populations in the Czech Republic from 2001 to 2010. A total of 398 P. cubensis isolates collected from Cucumis (Cm.) sativus, Cm. melo, Cucurbita (Cr.) maxima, Cr. pepo, Cr. moschata and Citrullus lanatus were analysed for variation in virulence (pathotypes). Virulence was evaluated on a differential set of 12 genotypes of cucurbitaceous plants. All isolates of P. cubensis were characterized by their level of virulence (classified according the number of virulence factors, VF; low VF = 1–4, medium VF = 5–8, high VF = 9–12): high (75%), medium (24%) and low (1%). The structure and dynamics of virulence in the pathogen populations were expressed by pathotypes using tetrad numerical codes and a total of 67 different pathotypes of P. cubensis were determined. The most susceptible group of differentials was Cucumis spp., while the lowest frequency of virulence was recorded on Cr. pepo ssp. pepo, Ci. lanatus and Luffa cylindrica. A high proportion (c. 90%) of isolates was able to infect cucurbit species Benincasa hispida and Lagenaria siceraria, which are not commonly cultivated in the Czech Republic or elsewhere in central Europe. In the recent pathogen populations (2008–2010) there was prevailing frequency (70–100%) of isolates with high numbers (9–12) of virulence factors. ‘Super pathotype’ 15.15.15 was often observed in the study within the pathogen populations and was one of the four most frequently recorded pathotypes. Pseudoperonospora cubensis populations shifted to a higher virulence over time. From 2009 the pathogen population changed dramatically and new pathotypes appeared able to establish natural and serious infection of Cucurbita spp. and Ci. lanatus, which was not observed in 2001–2008. Generally, virulence structure and dynamics of P. cubensis populations are extremely variable in the Czech Republic.     

The highly toxic Aconitum carmichaelii Debeaux as a root vegetable in the Qinling Mountains (Shaanxi, China)

Aconitum spp. are highly toxic due to the presence of aconitine-type alkaloids. Aconitum carmichaelii Debeaux is one of the most important and most studied Chinese medicines. However the scientific literature contains only anecdotal references to the use of A. carmichaelii as a food plant. The aim of the study was to document the traditional alimentary use of the species. Fifty people from three adjacent villages on Mount Taibai of the Qinling range (Shaanxi) were interviewed about the local use of the Aconitum. Most of them eat it on a regular basis every winter. The plant is widely cultivated in the area for food and nearly every family grows it. The tubers are cooked a few times during winter. Usually around 5 kg are prepared at one time and boiled, adding water four times, each time for at least 2 h, over a high flame. People usually eat one bowl a day (ca. 200 g). Respondents claim they eat them for two reasons: they heat the body in winter (they are “hot by nature”) and they are nourishing like other staples.     

Cryopreservation of ram semen: Manual versus programmable freezing and different lengths of equilibration

The aim of our study was to examine effects of the length of semen equilibration as well as two freezing techniques on ram sperm post-thaw quality. The ejaculates of Wallachian sheep rams (n = 12) were collected by an electro-ejaculation, equilibrated in a Triladyl® (0, 2, 4, 6, and 8 h) containing glycerol and egg yolk and frozen by programmable freezing (PF) or manual freezing (MF). After thawing, sperm samples were subjected to the motility (computer-assisted sperm analysis [CASA]), viability (SYBR-14/PI), and fertilizing ability (FA) (in vitro penetration/fertilization test on bovine oocytes) assays. It was found that the equilibration of 6 h (E-6) ensured higher post-thaw sperm motility and progressive movement compared with other lengths tested, irrespective of a freezing technique. The E-6 sperm viability did not differ between PF and MF but was lower (P < 0.05) than control. Sperm FA (E-6) was similar in PF (60.44%) and MF (62%) but slightly lower than in fresh (72.8%). Our data demonstrate that the use of MF was comparable with PF, which can be applied in the field conditions without need in a piece of cost-expensive equipment, which can greatly benefit the gene bank of animal genetic resources.     

Effects of granivorous rodents on direct seeding of oak and beech in relation to site preparation and sowing date

Planting bare-rooted seedlings is a rather expensive method to regenerate oak (Quercus robur) and beech (Fagus sylvatica). Direct seeding could reduce artificial regeneration costs substantially, but the method currently suffers from a number of problems, one of which is seed consumption by rodents. In the present study clear-cut areas in southern Sweden were prepared using one of four different mechanical site preparation methods – disc trenching, patch scarification, topsoil removal or mounding – then manually sown with acorns and beech nuts. Acorns were sown in May and July 2006, and both acorns and beech nuts were sown in May 2007. Three years following sowing, best seedling establishment from viable acorns, 60–70%, was achieved after sowing in mounds in May, and the subsequent survival reached 90–100%. After the first growing season for beech, both seedling establishment and subsequent survival was lower than for oak, which coincides with a higher number of lost beech nuts at the excavation to investigate the fate of seeds. Granivorous rodents (Apodemus flavicollis, A. sylvaticus, Mus musculus, and Myodes glareolus) were live-trapped more frequently in the vicinity of slash piles compared to other micro-site features, such as uprooted stumps, snags and stones. In 2006, no connection was found between number of rodents captured and proportion of lost seeds, while in 2007 the two were in good accordance. No clear relationship was found between number of captured rodents, vegetation cover class and mechanical site preparation method. More rodents were captured in 2007 than in 2006, and there was an increase in captures from spring until late summer/autumn for both years, which was mirrored in the seedling establishment result from the three different times of sowing. Based on experiences from the present study the recommendation for successful direct seeding of oak on clear-cuts is a combination of mounding site preparation, removal of slash and sowing in May rather than July. However, the results indicate that successful direct seeding of beech is more difficult to achieve.     

Porcine interleukin 2: parameters of production and biochemical characterization

Interleukin 2 (IL2) or T cell growth factor (TCGF) has been characterized in a number of species but not in porcines. Porcine IL2 was detected in supernates (SN) of cultures of pig lymphocytes by: 1) the stimulation of the IL2-sensitive murine T cell line, CT6; 2) a costimulator assay involving porcine thymocytes; and 3) by the in vitro maintenance of antigen or mitogen-induced porcine lymphoblastoid cells. Porcine IL2 production by pig lymphocytes was induced by the mitogens Concanavalin A (Con A) Phytohemagglutiniin (PHA), and Pokeweed mitogen (PWM), but not by lipopolysaccharide (LPS). IL2 activity was demonstrated in the SN of mitogen-stimulated lymphocyte cultures as early as 24 hr after initiation of culture, reached peak levels at 48 hr, and decreased by 72 hr. Mitogens induced IL2 secretion by pig peripheral blood mononuclear cells, lymph node cells, and spleen cells, but not thymus cells. The cells responsible for IL2 production are presumptive T cells because: 1) they are nylon wool non-adherent; and 2) are non-surface-Ig bearing. In contrast, SN from cultures of surface Ig-positive cells had minimal IL2 activity. Porcine IL2 resembles rat and human IL2 in that it has an apparent molecular weight of approximately 15,000, and does not bind to DEAE-cellulose (DE-52) ion exchange columns equilibrated in 0.05 M sodium phosphate buffer (pH 7.6).     

Chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence,under half of the adaptive growth-irradiance,for high-throughput sensing of leaf-water deficit in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> accessions

Background

Non-invasive and high-throughput monitoring of drought in plants from its initiation to visible symptoms is essential to quest drought tolerant varieties. Among the existing methods, chlorophyll a fluorescence (ChlF) imaging has the potential to probe systematic changes in photosynthetic reactions; however, prerequisite of dark-adaptation limits its use for high-throughput screening.

Results

To improve the throughput monitoring of plants, we have exploited their light-adaptive strategy, and investigated possibilities of measuring ChlF transients under low ambient irradiance. We found that the ChlF transients and associated parameters of two contrasting Arabidopsis thaliana accessions, Rsch and Co, give almost similar information, when measured either after ~20 min dark-adaptation or in the presence of half of the adaptive growth-irradiance. The fluorescence parameters, effective quantum yield of PSII photochemistry (Φ_PSII) and fluorescence decrease ratio (R _FD) resulting from this approach enabled us to differentiate accessions that is often not possible by well-established dark-adapted fluorescence parameter maximum quantum efficiency of PSII photochemistry (F _V/F _M). Further, we screened ChlF transients in rosettes of well-watered and drought-stressed six A. thaliana accessions, under half of the adaptive growth-irradiance, without any prior dark-adaptation. Relative water content (RWC) in leaves was also assayed and compared to the ChlF parameters. As expected, the RWC was significantly different in drought-stressed from that in well-watered plants in all the six investigated accessions on day-10 of induced drought; the maximum reduction in the RWC was obtained for Rsch (16%), whereas the minimum reduction was for Co (~7%). Drought induced changes were reflected in several features of ChlF transients; combinatorial images obtained from pattern recognition algorithms, trained on pixels of image sequence, improved the contrast among drought-stressed accessions, and the derived images were well-correlated with their RWC.

Conclusions

We demonstrate here that ChlF transients and associated parameters measured even in the presence of low ambient irradiance preserved its features comparable to that of measured after dark-adaptation and discriminated the accessions having differential geographical origin; further, in combination with combinatorial image analysis tools, these data may be readily employed for early sensing and mapping effects of drought on plant’s physiology via easy and fully non-invasive means.

     

Modeling population connectivity by ocean currents,a graph-theoretic approach for marine conservation

The dispersal of individuals among marine populations is of great importance to metapopulation dynamics, population persistence, and species expansion. Understanding this connectivity between distant populations is key to their effective conservation and management. For many marine species, population connectivity is determined largely by ocean currents transporting larvae and juveniles between distant patches of suitable habitat. Recent work has focused on the biophysics of marine larval dispersal and its importance to population dynamics, although few studies have evaluated the spatial and temporal patterns of this potential dispersal. Here, we show how an Eulerian advection–diffusion approach can be used to model the dispersal of coral larvae between reefs throughout the Tropical Pacific. We illustrate how this connectivity can be analyzed using graph theory—an effective approach for exploring patterns in spatial connections, as well as for determining the importance of each site and pathway to local and regional connectivity. Results indicate that the scale (average distance) of dispersal in the Pacific is on the order of 50–150 km, consistent with recent studies in the Caribbean (Cowen, et al. 2006). Patterns in the dispersal graphs highlight pathways for larval dispersal along major ocean currents and through island chains. A series of critical island ‘stepping stones’ are discovered providing potential pathways across the equatorial currents and connecting distant island groups. Patterns in these dispersal graphs highlight possible pathways for species expansions, reveal connected upstream/downstream populations, and suggest areas that might be prioritized for marine conservation efforts.     

泽泻蕨的系统位置——来自rbcL序列的分析

泽泻蕨的系统位置一直存有争议。该文根据叶绿体基因rbcL序列,用最大似然法构建它的系统发育关系。结果表明:①4种美洲典型的泽泻蕨属成员聚为一个很强支持的支,并与泽泻蕨形成姊妹群的关系,说明将泽泻蕨从泽泻蕨属中独立出来,放入拟泽泻蕨属更为合适。②泽泻蕨被聚在碎米蕨群分支,得到强烈支持。它与银粉背蕨和黑心蕨的关系比与拟金毛裸蕨属的关系更为近缘,不支持秦仁昌将泽泻蕨置于裸子蕨科的分类处理。③同意Panigrahi的观点,泽泻蕨的正确名称应为Parahemionitis cordata (Hooker&Greville) Fraser-Jenkins。     

Evaluation of the ability of colistin,amoxicillin (components of Potencil®), and fluoroquinolones to attenuate bacterial endotoxin‐ and Shiga exotoxin‐mediated cytotoxicity—In vitro studies

Escherichia coli is one of the major pathogens in humans and animals causing localized and systemic infections, which often lead to acute inflammation, watery diarrhea, and hemorrhagic colitis. Bacterial lipopolysaccharide (LPS) and Shiga exotoxins (Stx) are mostly responsible for such clinical signs. Therefore, highly effective treatment of E. coli infections should include both eradication of bacteria and neutralization of their toxins. Here, for the first time, we compared the in vitro ability of common antibiotics to decrease LPS‐ and Stx‐mediated cytotoxicity: colistin, amoxicillin (used separately or combined), enrofloxacin, and its metabolite ciprofloxacin. Three experimental scenarios were realized as follows: (a) the direct effect of antibiotics on endotoxin, (b) the effect of antibiotic treatment on LPS‐mediated cytotoxicity in an experiment mimicking “natural infection,” (c) the effect of antibiotics to decrease Stx2e‐mediated cytotoxicity. Two cell lines, A549 and Vero cells, were used to perform cytotoxic assays with the methyl tetrazolium (MTT) and lactate dehydrogenase leakage (LDH) methods, respectively. Colistin and amoxicillin, especially used in combination, were able to attenuate LPS toxic effect, which was reflected by increase in A549 cell viability. In comparison with other antibiotics, the combination of colistin and amoxicillin exhibited the highest boster or additive effect in protecting cells against LPS‐ and Stx2e‐induced toxicity. In summary, in comparison with fluoroquinolones, the combination of colistin and amoxicillin at concentrations similar to those achieved in plasma of treated animals exhibited the highest ability to attenuate LPS‐ and Stx2e‐mediated cytotoxicity.     

AIDS, drought, and child malnutrition in southern Africa

OBJECTIVE: To investigate trends in child malnutrition in six countries in southern Africa, in relation to the HIV epidemic and drought in crop years 2001/2 and 2002/3. DESIGN: Epidemiological analysis of sub-national and national surveys with related data.Setting: Data from Lesotho, Malawi, Mozambique, Swaziland, Zambia and Zimbabwe, compiled and analysed under UNICEF auspices. SUBJECTS: Secondary data: children 0-5 years for weight-for-age; HIV prevalence data from various sources especially antenatal clinic surveillance. RESULTS: Child nutritional status as measured by prevalence of underweight deteriorated from 2001 onwards in all countries except Lesotho, with very substantial increases in some provinces/districts (e.g. from 5 to 20% in Maputo (Mozambique, 1997-2002), 17 to 32% in Copperbelt (Zambia, 1999-2001/2) and 11 to 26% in Midlands province (Zimbabwe, 1999-2002)). Greater deterioration in underweight occurred in better-off areas. Areas with higher HIV/AIDS prevalences had (so far) lower malnutrition rates (and infant mortality rates), presumably because more modern areas--with greater reliance on trade and wage employment--have more HIV/AIDS. Areas with higher HIV/AIDS showed more deterioration in child nutrition. A significant area-level interaction was found of HIV/AIDS with the drought period, associated with particularly rapid deterioration in nutritional status. CONCLUSIONS: First, the most vulnerable may be households in more modern areas, nearer towns, to whom resources need to be directed. Second, the causes of this vulnerability need to be investigated. Third, HIV/AIDS amplifies the effect of drought on nutrition, so rapid and effective response will be crucial if drought strikes again. Fourth, expanded nutritional surveillance is now needed to monitor and respond to deteriorating trends. Finally, with or without drought, new means are needed of bringing help, comfort and assistance to the child population.