A simplified procedure for terminal restriction fragment length polymorphism analysis of the soil bacterial community to study the effects of pesticides on the soil microflora using 4,6-dinitroorthocresol as a test case

We have studied the structural effects of application to the soil of a potentially detrimental herbicide, 4,6-dinitroorthocresol (DNOC) by analysing amplified ribosomal DNA restriction analysis (ARDRA) and terminal restriction fragment length polymorphism (T-RFLP) signatures of 16S rDNA fragments of culturable bacterial communities isolated from diluted soil suspensions. This approach has the potential to reveal changes induced by stressing the soil microflora with DNOC. This paper shows that, whereas only few changes of the ARDRA and T-RFLP profiles result from ageing of the soil, treatment of the soil with DNOC induces major modifications of these profiles. Therefore, for the practical purpose of pesticide registration, ARDRA and T-RFLP analysis performed on the dominant culturable fraction of the soil bacteria, implemented using conventional gel electrophoresis, offers the means of a routine, simple and meaningful test for detecting some of the changes affecting the structure of the soil microflora in response to pesticide application.     

Closed-shell molecules that ionize more readily than cesium

We report a class of molecules with extremely low ionization enthalpies, one member of which has been determined to have a gas-phase ionization energy (onset, 3.51 electron volts) lower than that of the cesium atom (which has the lowest gas-phase ionization energy of the elements) or of any other known closed-shell molecule or neutral transient species reported. The molecules are dimetal complexes with the general formula M2(hpp)4 (where M is Cr, Mo, or W, and hpp is the anion of 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]pyrimidine), structurally characterized in the solid state, spectroscopically characterized in the gas phase, and modeled with theoretical computations. The low-energy ionization of each molecule corresponds to the removal of an electron from the delta bonding orbital of the quadruple metal-metal bond, and a strong interaction of this orbital with a filled orbital on the hpp ligands largely accounts for the low ionization energies.     

The proteasome of Mycobacterium tuberculosis is required for resistance to nitric oxide

The production of nitric oxide and other reactive nitrogen intermediates (RNI) by macrophages helps to control infection by Mycobacterium tuberculosis (Mtb). However, the protection is imperfect and infection persists. To identify genes that Mtb requires to resist RNI, we screened 10,100 Mtb transposon mutants for hypersusceptibility to acidified nitrite. We found 12 mutants with insertions in seven genes representing six pathways, including the repair of DNA (uvrB) and the synthesis of a flavin cofactor (fbiC). Five mutants had insertions in proteasome-associated genes. An Mtb mutant deficient in a presumptive proteasomal adenosine triphosphatase was attenuated in mice, and exposure to proteasomal protease inhibitors markedly sensitized wild-type Mtb to RNI. Thus, the mycobacterial proteasome serves as a defense against oxidative or nitrosative stress.     

Essential role of Fkbp6 in male fertility and homologous chromosome pairing in meiosis

Meiosis is a critical stage of gametogenesis in which alignment and synapsis of chromosomal pairs occur, allowing for the recombination of maternal and paternal genomes. Here we show that FK506 binding protein (Fkbp6) localizes to meiotic chromosome cores and regions of homologous chromosome synapsis. Targeted inactivation of Fkbp6 in mice results in aspermic males and the absence of normal pachytene spermatocytes. Moreover, we identified the deletion of Fkbp6 exon 8 as the causative mutation in spontaneously male sterile as/as mutant rats. Loss of Fkbp6 results in abnormal pairing and misalignments between homologous chromosomes, nonhomologous partner switches, and autosynapsis of X chromosome cores in meiotic spermatocytes. Fertility and meiosis are normal in Fkbp6 mutant females. Thus, Fkbp6 is a component of the synaptonemal complex essential for sex-specific fertility and for the fidelity of homologous chromosome pairing in meiosis.     

Biotechnische Regulierung der Kohlmottenschildlaus in Kohlgem��se �C Sachstand und Perspektiven

The cabbage whitefly,Aleyrodes proletella, has developed to a key pest in brassica vegetable crops throughout Germany. In a 3-year project we investigated a biotechnical approach employing netting (0.8?×?0.8 mm meshsize) in combination with the native parasitoidEncarsia tricolor in an inoculative-inundative approach in organic Brussels sprouts under field conditions. In small scale plot experiments continuous netting from transplanting in May till October alone reduced whitefly larval densities by 77 % at peak infestation in all years. Remaining whitefly infestation under net was associated with the necessity of temporary net removal for the purpose of mechanical weeding, which caused primary whitefly infestation. Release of mass rearedE. tricolour under net at the first signs of larval infestation significantly increasedA. proletella-parasitation and reduced infestation compared to netted controls without release. Furthermore, at highest dosage in 2009,E. tricolor had a significant enhancing effect on raw yield and the quality level of marketable yield. Additional on-farm experiments without netting in 2008 and 2009 confirmed the control potential ofE. tricolor. Although the most important project mile stones have been achieved, namely (a)E. tricolour-establishment under net and in the open field, (b) enhanced parasitation corresponding with pest density decline and (c) a positive impact on yield and quality parameters, further research will be necessary. The focus should be on the initial infestation period, optimizing the timing, frequency and required dosage.     

A retrospective study of 11 cases of lungworm (Didelphostrongylus hayesi) infection in opossums (Didelphis virginiana).

A juvenile, female North American opossum (Didelphis virginiana) died of verminous pneumonia caused by Didelphostrongylus haysei despite aggressive treatment with oral fenbendazole, corticosteroids, and antibiotics. This prompted a retrospective study of lungworm infection in opossums, during which 19 additional necropsy reports from opossums were reviewed. Including the subject of this report, a total of 11 (55%) of these cases included a diagnosis of lungworm infection. This diagnosis was considered to have contributed to death in eight out of the 11 cases (73%). Histologically, 10 of the 11 (91%) opossums had granulomatous bronchopneumonia with small to moderate numbers of adult nematodes in the airways and parenchyma. Four of the 11 (36%) opossums had free larvae within the parenchyma or terminal airways. Inflammation was usually associated with larvae, degenerating parasites, and nonintact adult nematodes. Superimposed bacterial pneumonia was evident in three animals, and sections of lung examined from all the opossums were characterized by moderate to severe smooth-muscle hyperplasia in airways, including terminal respiratory bronchioles and alveolar ducts. Nine animals had prominent medial smooth-muscle hyperplasia in small- and medium-sized arterioles. Lesions in other organs, particularly in liver, heart, and gastrointestinal tract, were frequently identified. Three animals had concomitant septicemia or bacterial bronchopneumonia (or both), which contributed to the cause of death. Seven animals had gastric nematodosis (Physaloptera sp.), although three of them had been treated with a 14-day course of fenbendazole.     

Salmonella enterica serovar Enteritidis colonization of the chicken caecum requires the HilA regulatory protein

Invasion of Salmonella into intestinal epithelial cells is believed to be essential for the pathogenesis of Salmonella infections. Invasion is mediated by genes located on the Salmonella pathogenicity Island I (SPI-1), which are needed for assembling a type three secretion system, that mediates injection of bacterial proteins into the cytosol of epithelial cells, resulting in cytoskeletal rearrangements and as a consequence invasion. HilA is the key regulator of the Salmonella Pathogenicity Island I. To assess the role of hilA in colonization of gut and internal organs in poultry, animals were infected with 10(8) CFU of a delta hilA mutant of S. Enteritidis and its parent strain at day of hatch. Very low numbers of delta hilA mutant strain were able to colonize the internal organs shortly after infection, but they were not eliminated from internal organs at 4 weeks post-infection. At that time, the colonization level of the wild type bacteria in internal organs was decreased to the same low level compared with delta hilA mutant strain bacteria. Shedding of the delta hilA mutant strain and colonization of the caeca was seriously decreased relative to the parent strain starting from Day 5 post-infection. At 4 weeks post-infection, the delta hilA mutant strain was more or less eliminated from the chicken gut, while the parent strain was still shed to a high level and colonized the caeca to a high extent (more than 10(7) CFU/g). It is concluded that hilA is involved in long-term shedding and colonization of S. Enteritidis in the chicken caeca.     

Understanding Variability in Adaptive Capacity on Rangelands

The art and science of developing effective policies and practices to enhance sustainability and adapt to new climate conditions on rangelands and savannas are typically founded on addressing the “average” or “typical” resource user. However, this assumption is flawed since it does not appreciate the extent of diversity among resource users; it risks that strategies will be irrelevant for many people and ignored, and that the grazing resource itself will remain unprotected. Understanding social heterogeneity is vital for effective natural resource management. Our aim was to understand the extent to which graziers in the northern Australian rangelands varied in their capacity to adapt to climate variability and recommended practices. Adaptive capacity was assessed according to four dimensions: 1) the perception of risk, 2) skills in planning, learning and reorganising, 3) financial and emotional flexibility, and 4) interest in adapting. We conducted 100 face-to-face interviews with graziers in their homes obtaining a 97% response rate. Of the 16 possible combinations that the four dimensions represent, we observed that all combinations were present in the Burdekin. Any single initiative to address grazing land management practices in the region is unlikely to address the needs of all graziers. Rather, policies could be tailored to type-specific needs based on adaptive capacity. Efforts to shift graziers from very low, low, or moderate levels of adaptive capacity are urgently needed. We suggest some strategies.     

Molecular epidemiology of Mycobacterium avium subsp. avium and Mycobacterium intracellulare in captive birds

Mycobacterium avium subsp. avium and Mycobacterium intracellulare are primary causes of mycobacteriosis in captive birds throughout the world, but little is known about how they are transmitted. To define the local epidemiology of infection, we strain-typed 70 M. avium subsp. avium and 15 M. intracellulare culture isolates obtained over a 4-year period from captive birds. Typing was performed using randomly amplified polymorphic DNA (RAPD) PCR, amplified fragment length polymorphic (AFLP) fragment analyses, and for a subset of isolates, DNA sequencing of a segment of the 16S-23S rRNA internal transcribed spacer region. Six strain clusters comprising 43 M. avium subsp. avium, isolates were identified; 42 isolates had unique typing patterns, including all M. intracellulare isolates. Phylo-geographical analyses using RAPD and AFLP fingerprints and animal confinement histories showed no correlation between housing of infected birds and mycobacterial strain-type, except for two animals. The diversity of M. avium subsp. avium and M. intracellulare isolates and minimal evidence for bird-to-bird transmission suggest that environmental reservoirs may be important sources of infection in captivity.