Molecular markers for detection of pathogenic Escherichia coli strains belonging to serogroups O 138 and O 139

Escherichia coli strains belonging to O-serogroup 138 and 139 are important as disease agents in pigs causing post-weaning diarrhea and edema disease. Several types of shiga toxin-producing O 138 and O 139 strains were isolated from diarrheic humans and from cattle and food of bovine origin. Serotyping is the current method for detection of O 138 and O 139 strains but its applicability can be limited due to the presence of capsules and capsular-like bacterial surface antigens and in the case of rough LPS. To overcome these difficulties for diagnosis, we have developed a specific PCR method suitable for detection of different types of O 138 and O 139 strains. The O-antigen gene clusters of E. coli O 138 and O 139 type strains were sequenced, and the genes were identified on the basis of homology. By screening against 186 E. coli and Shigella type strains, two genes specific to each of E. coli O 138 and O 139 were identified, respectively, and were tested on 15 clinical and environmental isolates of those two serogroups in a double-blind test. The sensitivity of the PCR assays was determined, and the detection limits were 2 pg per mul of chromosomal DNA and 2 CFU per 10 g of water or pork samples. PCR-based detection of O-antigen specific genes of E. coli O 138 and O 139 was shown to be accurate, highly sensitive and rapid, and is suggested as a new diagnostic tool for investigations of infections and outbreaks with these strains in animals and humans and for control of food.     

Functional risk assessment of handling of sheep housing and pasture areas at a scrapie-infested farm in Mecklenburg-West Pomerania

In the wake of a scrapie monitoring programme 2002 in Mecklenburg-Westpomerania a total of 10 positive cases were identified after culling at a suffolk-farm with about 1000 animals. Scientific based recommendations are difficult to judge as longitudinal studies and subsequent quantitative risk factors regarding the epidemiology of scrapie are either not available or conflicting. Given this background a risk assessment was developed that is based on the probability of validated risk factors considering special husbandry and circumstances at the very farm.     

Comparative analysis of neonicotinoid binding to insect membranes: I. A structure-activity study of the mode of [3H]imidacloprid displacement in Myzus persicae and Aphis craccivora

Neonicotinoids bind selectively to insect nicotinic acetylcholine receptors with nanomolar affinity to act as potent insecticides. While the members of the neonicotinoid class have many structural features in common, it is not known whether they also share the same mode of binding to the target receptor. Previous competition studies with [3H]imidacloprid, the first commercialised neonicotinoid, indicated that thiamethoxam, representing a novel structural sub-class, may bind in a different way from that of other neonicotinoids. In the present work we analysed the mode of [3H]imidacloprid displacement by established neonicotinoids and newly synthesized analogues in the aphids Myzus persicae Sulzer and Aphis craccivora Koch. We found two classes of neonicotinoids with distinct modes of interference with [3H]imidacloprid, described as direct competitive inhibition and non-competitive inhibition, respectively. Competitive neonicotinoids were acetamiprid, nitenpyram, thiacloprid, clothianidin and nithiazine, whereas thiamethoxam and the N-methyl analogues of imidacloprid and clothianidin showed non-competitive inhibition. The chloropyridine or chlorothiazole heterocycles, the polar pharmacophore parts, such as nitroimino, cyanoimino and nitromethylene, and the cyclic or acyclic structure of the pharmacophore were not relevant for the mode of inhibition. Consensus structural features of the neonicotinoids were defined for the two mechanisms of interaction with [3H]imidacloprid binding. Furthermore, two sub-classes of non-competitive inhibitors can be discriminated on the basis of their Hill coefficients for imidacloprid displacement. We conclude from the present data that the direct competitors share the binding site with imidacloprid, whereas non-competitive compounds, like thiamethoxam, bind to a different site or in a different mode.     

Comparative analysis of neonicotinoid binding to insect membranes: II. An unusual high affinity site for [3H]thiamethoxam in Myzus persicae and Aphis craccivora

Neonicotinoids represent a class of insect-selective ligands of nicotinic acetylcholine receptors. Imidacloprid, the first commercially used neonicotinoid insecticide, has been studied on neuronal preparations from many insects to date. Here we report first intrinsic binding data of thiamethoxam, using membranes from Myzus persicae Sulzer and Aphis craccivora Koch. In both aphids, specific binding of [3H]thiamethoxam was sensitive to temperature, while the absolute level of non-specific binding was not affected. In M persicae, binding capacity (Bmax) for [3H]thiamethoxam was ca 450 fmol mg(-1) of protein at 22 degrees C and ca 700 fmol mg(-1) of protein at 2 degrees C. The negative effect of increased temperature was reversible and hence not due to some destructive process. The affinity for [3H]thiamethoxam was less affected by temperature: Kd was ca 11 nM at 2 degrees C and ca 15 nM at 22 degrees C. The membranes also lost binding sites for [3H]thiamethoxam during prolonged storage at room temperature, and upon freezing and thawing. In A craccivora, [3H]thiamethoxam was bound with a capacity of ca 1000 fmol mg(-1) protein and an affinity of ca 90 nM, as measured at 2 degrees C. Overall, the in vitro temperature sensitivity of [3H]thiamethoxam binding was in obvious contrast to the behaviour of [3H]imidacloprid studied in parallel. Moreover, the binding of [3H]thiamethoxam was inhibited by imidacloprid in a non-competitive mode, as shown with M persicae. In our view, these differences demonstrate that thiamethoxam and imidacloprid, which represent different structural sub-classes of neonicotinoids, do not share the same binding site or mode. This holds also for other neonicotinoids, as we report in a companion article.     

Pymetrozine,a Fast-Acting and Selective Inhibitor of Aphid Feeding. In-situ Studies with Electronic Monitoring of Feeding Behaviour

Pymetrozine, a pyridine azomethine compound, represents a novel insecticide with a selective activity against homopteran insects. It acts in a unique way: aphids are not knocked down on contact but seem to die of starvation. This implies an effect of pymetrozine on feeding behaviour. The aim of the present work was to elucidate how and at which step pymetrozine interferes with the complex mechanisms underlying phloem feeding. The effect of pymetrozine, applied in various ways, on different phases of stylet penetration and feeding activity of individual aphids was studied using the Electrical Penetration Graph technique (EPG). Initial choice experiments indicated that pymetrozine does not have a deterrent or antifeedant action. Topical application (150 ng pymetrozine mg^-1 fresh weight) inhibited stylet insertion into the plant. When injected, less than 30 ng mg^-1 was sufficient to produce the same effect. When pymetrozine was systemically applied via plant spraying or root uptake, aphids started feeding normally. After some time, however, they withdrew their stylets from the phloem and walked around with unaffected locomotion. At low doses aphids eventually recovered and resumed feeding. High doses, however, irreversibly disrupted feeding and prevented stylet reinsertion. Aphid motility was not affected up to an estimated haemolymph concentration of 1 mM pymetrozine. Aphids which eventually stopped feeding on pymetrozine-treated plants showed EPGs with distorted salivation/ingestion patterns. It is concluded that pymetrozine does not have a general toxic effect on aphids but selectively interferes with the nervous regulation of feeding behaviour which consequently results in death due to starvation after a few days. © 1997 SCI.     

Control and measurement of three-qubit entangled states

We report the deterministic creation of maximally entangled three-qubit states-specifically the Greenberger-Horne-Zeilinger (GHZ) state and the W state-with a trapped-ion quantum computer. We read out one of the qubits selectively and show how GHZ and W states are affected by this local measurement. Additionally, we demonstrate conditional operations controlled by the results from reading out one qubit. Tripartite entanglement is deterministically transformed into bipartite entanglement by local operations only. These operations are the measurement of one qubit of a GHZ state in a rotated basis and, conditioned on this measurement result, the application of single-qubit rotations.     

Structure of the light-driven chloride pump halorhodopsin at 1.8 A resolution

Halorhodopsin, an archaeal rhodopsin ubiquitous in Haloarchaea, uses light energy to pump chloride through biological membranes. Halorhodopsin crystals were grown in a cubic lipidic phase, which allowed the x-ray structure determination of this anion pump at 1.8 angstrom resolution. Halorhodopsin assembles to trimers around a central patch consisting of palmitic acid. Next to the protonated Schiff base between Lys(242) and the isomerizable retinal chromophore, a single chloride ion occupies the transport site. Energetic calculations on chloride binding reveal a combination of ion-ion and ion-dipole interactions for stabilizing the anion 18 angstroms below the membrane surface. Ion dragging across the protonated Schiff base explains why chloride and proton translocation modes are mechanistically equivalent in archaeal rhodopsins.     

On the Isotopic Composition of Pb in Cloud Waters in Central Germany. A Source Discrimination Study

Fifty seven (57) cloud water samples collected in 1 hr intervals during 9cloud events in 1997 at the summit of the Brocken (altitude 1142 m) in Central Germany were analyzed for their Pb concentrations and isotopic compositions. For comparison, filterdusts of such possible emitters as power plants, cement factories, waste incinerators, cars and others were also studied.Profiles over many hours reveal changes in composition correlatedwith changes in the origin of the air masses. Five main groups ofpossible sources of the Pb can be distinguished: most importantare (1) the European standard pollution ESP dominating the continent from Germany over France to Scandinavia and (2) industrial Pb (and soot from car exhausts) followed by (3) urban waste incinerators as well as (4) perhaps ores from Norilsk/Siberia, and, somewhat uncertain, (5) ores from PaleozoicEuropean deposits. Local and daily varying anthropogenic emissions of the sources (2–5) cause scatter of the points around the ESP correlation line. In soils all these variationsand individual signatures are levelled out so that specific sources of pollution cannot be identified anymore. Geogenic Pbfrom rocks and soils is the same as that from power plants and cement factories. It is one of the two nominal end components making up the mixture of the ESP. The less radiogenic end component of this mix which must contain Pb from very old deposits is somewhat elusive: none of the large Archaic or Proterozoic deposits could be identified. Most probably numerous sources contribute to this Pb. Pb from leaded gasoline seems to be absent or is hidden under the five components dominating the composition of the Pb in the European environment. This probably reflects phasing out of such gasoline in most European countries. The Pb in the clouds may contain a small component of old Proterozoic or even Archaic Pb, but it can also be modelled with Paleozoic Pb.     

The Isotopic Composition of Anthropogenic Pb in Soil Profiles of Northern Germany: Evidence for Pollutant Pb from a Continent-wide Mixing System

Seventy-five samples of six soil profiles from forests on different substrates plus one next to a motorway were analyzed for the variation of Pb concentration and isotopic composition with depth. A substantial fraction of the anthropogenic Pb (peak concentrations between 74 and 300 ppm) is still stored in the organic top soil, and seldom penetrates deeper than 20–30 cm. In (²⁰⁸Pb/²⁰⁶Pb) vs. (²⁰⁷Pb/²⁰⁶Pb)-diagrams the Pb in the uppermost layers of the profiles plots on an excellent correlation line (R = 0.99) indicating essentially a two-component mixing system. Values for the dust on Scottish plants, Norwegian and Swiss bogs as well as for aerosols collected over the North Atlantic (data from the literature) plot on the same correlation line showing that the Pb was deposited from a continent-wide mixing system with two apparent end members: The radiogenic one is similar to geogenic Pb and could consist of contributions from certain European ore deposits and from different soil components (e.g. silicates and Fe-oxides), power plants and cement factories. The less radiogenic component must contain or consist of Pb from Proterozoic or even Archaic ore deposits. However, the sources of this Pb remain somewhat elusive. In any case, this isotopic pattern is so consistent all over Europe that the correlation line may serve as a line of reference for quasi normal or European Standard Lead Pollution (ESLP). The gasoline derived Pb in the soils is swamped by Pb of other provenances.