Molecular detection of sugarcane striate virus and sugarcane white streak virus and their prevalence in the Miami World Collection of sugarcane and related grasses

Viruses in the genus Mastrevirus (family Geminiviridae), including those infecting sugarcane, have natural geographical ranges almost exclusively restricted to Africa and the Indian Ocean islands off the African coast. Only sugarcane white streak virus (SWSV) in Barbados and sugarcane striate virus (SStrV) in Florida and Guadeloupe are known to infect a few sugarcane varieties in the Western Hemisphere. In this study, PCR assays were developed to detect these two viruses in sugarcane. Five hundred and seventy-one DNA samples from Saccharum species and interspecific hybrids from the Miami World Collection of sugarcane and related grasses were tested for the presence of SStrV and SWSV by PCR. No variety was found infected by SWSV but SStrV was detected in 19 varieties. PCR data were confirmed by sequencing amplified fragments (248 bp). These fragments shared 93%–100% nucleotide identity with SStrV sequences from the GenBank database. SStrV isolates were distributed in six phylogenetic groups, including the four strains of the virus. Most varieties infected by SStrV originated from Asia, thus confirming a previous hypothesis stating that this virus originated from this continent. Absence of SStrV in commercial sugarcane in Florida also suggested that this virus has not been spread in this location, while infected plants have been present for several decades.     

Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Fine‐scale habitat partitioning of Chilean and Peale's dolphins and their overlap with aquaculture

1. Predictive species distribution models (SDMs) have become powerful tools to determine habitat use patterns of mobile marine predators and their spatial overlap with potentially impacting anthropogenic activities.
2. This study used SDMs to investigate fine‐scale habitat use patterns of two poorly known and broadly sympatric coastal delphinids, Chilean dolphins (Cephalorhynchus eutropia) and Peale's dolphins (Lagenorhynchus australis), and their spatial interactions with intense aquaculture farming activities in the Chiloé archipelago, southern Chile.
3. A long‐term dataset (2002–2012) of boat‐based dolphin sightings and concurrently in situ collected environmental and anthropogenic variables was analysed using binomial Generalized Additive Models to investigate ecological drivers of each species' fine‐scale distribution and to predict dolphin occurrence spatially.
4. Chilean dolphins preferred shallow (<30 m deep), turbid waters, close to shore (<500 m) and river mouths which often placed them in sheltered bays and channels used intensively by shellfish farms. Peale's dolphins were also found in shallow waters but occurred over a wider range of conditions along more open or exposed coastlines. Both species had to navigate extensive salmon and shellfish farming sites to transit between areas of important habitat.
5. Sightings and predicted occurrence maps showed a clear pattern of spatial habitat partitioning between species, which remained stable across the 11 year study period. The identification of important habitat for Chilean dolphins warrants the consideration of spatially explicit conservation measures to limit the potential effects of overlapping salmon and shellfish farming.
6. The observed differences in ecological plasticity of the two sympatric species should be considered when evaluating and mitigating the effects of environmental change and ongoing anthropogenic pressures on their nearshore habitat. The estimated species–environment relationships could also be used to predict where dolphin habitat and anthropogenic activities are most likely to overlap in other parts of the species' ranges.

     

Zum Auftreten und zur Bekämpfung von Erdbeerschädlingen in Norddeutschland

Zusammenfassung Auf Grund seiner ausgezeichneten Tiefenwirkung und einer noch ausreichenden Dauerwirkung hat sich der Oleo-Phosphorsäureester in Versuchen gegen die Erdbeermilbe und in einem praktischen Einsaß gegen einen Erdbeerwickler als brauchbares Akarizid bzw. Insektizid erwiesen. Er wird daher auf Grund der weiteren guten Erfahrungen mit dieser Wirkstoffzubereitung im Kern- und Steinobstbau auch gegen andere wichtige Erdbeerschädlinge, insbesondere gegen Rote Spinne und Erdbeerblattlaus als wichtigen Virusüberträger, empfohlen. Der Oleo-Phosphorsäureester wird zu einer Art Universalmittel, wenn durch ihn auch noch Erdbeerstengel- und-blütenstecher erfaßt werden, vielleicht sogar auch parasitische Nematoden (Stengel- und Blattä lchen). Gegen freilebende (ektoparasitische) Nematoden werden vorerst weiterhin nur die Spezialmittel zur Bodenentseuchung in Betracht kommen.Eine einmalige Sprißung, etwa zehn Tage vor der Erdbeerblüte, mit Folidol-Öl 0,5%, 10001 je ha, und eine weitere Sprißung bald nach der Ernte mit 2000 I je ha werden daher im allgemeinen als ausreichend für die Bekämpfung tierischer Schädlinge im Erdbeeranbau betrachtet. Nur gegen die Blatt- und Stengelälchen wird noch eine zusäßliche Behandlung bei Blattaustrieb im Frühjahr notwendig sein.Hygienische Bedenken bestehen zunächst bei dem Einsaß des Oleo-Phosphorsäureesters vor der Erdbeerblüte nicht; in umfangreichen Testen konnte keine merkliche Geschmacksbeeinflussung durch den Wirkstoff festgestellt werden. Auch die Karenzzeit (etwa 14 Tage) dürfte bei rechtzeitigem Einsaß vor der Blüte mit mindestens 6 Wochen bis zur Ernte reichlich bemessen sein. Nachbarkulturen, die früh geerntet werden, z. B. Frühgemüse, dürfen vom Sprißstrahl oder von Abtrift selbstverständlich nicht betroffen werden.Thiodan wird als Mittel gegen die Erdbeermilbe und andere Erdbeerschädlinge kaum in Betracht kommen, da es bei einer Vorblütenspritzung eine schon merkliche Geschmacksbeeinflussung der Erdbeerfrüchte hinterlä ßt.     

Depletion of non‐exchangeable NH$ _4^+ $ at the root–soil and hyphae–soil interface of VA mycorrhizal maize (Zea mays) and parsley (Petroselinum sativum)

Mobilization of non‐exchangeable ammonium (NH ) by hyphae of the vesicular‐arbuscular mycorrhizal (VAM) fungus Glumus mosseae was studied under controlled experimental conditions. Maize (Zea mays) and parsley (Petroselinum sativum) were grown either alone or in symbiosis with Glomus mosseae in containers with separated compartments for roots and hyphal growth. In one experiment, ¹⁵NH was added to the soil to differentiate between the native non‐exchangeable NH and the non‐exchangeable NH derived from N fertilization. Non‐exchangeable NH was mobilized by plant growth. Plant dry weight and N uptake, however, were not significantly influenced by mycorrhizal colonization of the roots. The influence of root infection with mycorrhizal fungus on the mobilization of non‐exchangeable NH was negligible. In the hyphal compartment, hyphal uptake of N resulted in a decrease of NH in the soil solution and of exchangeable NH . However, the NH concentration was still too high to permit the release of non‐exchangeable NH . The results demonstrate that, in contrast to roots, hyphae of VAM fungi are not able to form a non‐exchangeable‐NH depletion zone in the adjacent soil. However, under conditions of a more substantial depletion of the exchangeable NH in the mycorrhizal sphere (e.g., with longer growth), an effect of mycorrhiza on the non‐exchangeable NH might be found.     

Sulfur in soils

Sulfur (S) deficiency of crops, which has been reported with increasing frequency over the past two decades on a worldwide scale, is a factor that reduces yield and affects the quality of harvested products. Especially in Western European countries, incidence of S deficiency has increasingly been reported in Brassicaceae. For this reason, more attention should be paid to the optimization of S‐fertilizer application, in order to cover plant S requirements whilst minimizing environmental impacts. In soils, S exists in inorganic and organic forms. While sulfate (SO ), which is a direct S source for plants, contributes up to 5% of total soil S, generally more than 95% of soil S are organically bound. Organic S is divided into sulfate ester and carbon‐bonded S. Although not directly plant‐available, organically bound S may potentially contribute to the S supply of plants, especially in deficiency situations. Sulfur turnover involves both biochemical and biological mineralization. Biochemical mineralization, which is the release of SO from the ester sulfate pool through enzymatic hydrolysis, is controlled by S supply, while the biological mineralization is driven by the microbial need for organic C to provide energy.     

Der Bulk-Stomata-Widerstand von Ackerbohnen und seine Abhängigkeit vom Bodenwassergehalt

Bulk stomatal resistance of Vicia faba and its dependence on soil water content Bulk stomatal resistance of beans (Vicia faba L.,) as a function of soil water content was estimated for 41 days by comparison of potential transpiration, determined from meteorological data, and actual transpiration, determined from soil water losses. The resistance of beans is about 46 s/m if soil water content is above 70 % of available field capacity and increases strongly in dryer situations. This behaviour can be described by an exponential function.     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

Fate of Cry1Ab protein in agricultural systems under slurry management of cows fed genetically modified maize (Zea mays L.) MON810: a quantitative assessment

The objective of the study was to track the fate of recombinant Cry1Ab protein in a liquid manure field trial when feeding GM maize MON810 to dairy cows. A validated ELISA was applied for quantification of Cry1Ab in the agricultural chain from GM maize plants, feed, liquid manure and soil to crops grown on manured fields. Starting with 23.7 μg of Cry1Ab g(-1) dry weight GM maize material, a rapid decline of Cry1Ab levels was observed as 2.6% and 0.9% of Cry1Ab from the GM plant were detected in feed and liquid manure, respectively. Half of this residual Cry1Ab persisted during slurry storage for 25 weeks. After application to experimental fields, final degradation of Cry1Ab to below detectable levels in soil was reported. Cry1Ab exhibited a higher rate of degradation compared to total protein in the agricultural processes. Immunoblotting revealed a degradation of the 65 kDa Cry1Ab into immunoreactive fragments of lower size in all analyzed materials.