Assessment of bacterial endosymbiont diversity in Otiorhynchus spp. (Coleoptera: Curculionidae) larvae using a multitag 454 pyrosequencing approach

<p>Abstract</p> <p>Background</p> <p>Weevils of the genus <it>Otiorhynchus</it> are regarded as devastating pests in a wide variety of horticultural crops worldwide. So far, little is known on the presence of endosymbionts in <it>Otiorhynchus</it> spp.. Investigation of endosymbiosis in this genus may help to understand the evolution of different reproductive strategies in these weevils (parthenogenesis or sexual reproduction), host-symbiont interactions, and may provide a future basis for novel pest management strategy development. Here, we used a multitag 454 pyrosequencing approach to assess the bacterial endosymbiont diversity in larvae of four economically important <it>Otiorhynchus</it> species.</p> <p>Results</p> <p>High-throughput tag-encoded FLX amplicon pyrosequencing of a bacterial <it>16S</it> rDNA fragment was used to characterise bacterial communities associated with different <it>Otiorhynchus</it> spp. larvae. By sequencing a total of ~48,000 PCR amplicons, we identified 49 different operational taxonomic units (OTUs) as bacterial endosymbionts in the four studied <it>Otiorhynchus</it> species. More than 90% of all sequence reads belonged either to the genus <it>Rickettsia</it> or showed homology to the phylogenetic group of “<it>Candidatus</it> Blochmannia” and to endosymbionts of the lice <it>Pedicinus obtusus</it> and <it>P. badii</it>. By using specific primers for the genera <it>Rickettsia</it> and “<it>Candidatus</it> Blochmannia”, we identified a new phylogenetic clade of <it>Rickettsia</it> as well as “<it>Candidatus</it> Nardonella” endosymbionts in <it>Otiorhynchus</it> spp. which are closely related to “<it>Candidatus</it> Blochmannia” bacteria.</p> <p>Conclusions</p> <p>Here, we used multitag 454 pyrosequencing for assessment of insect endosymbiotic communities in weevils. As 454 pyrosequencing generates only quite short sequences, results of such studies can be regarded as a first step towards identifying respective endosymbiotic species in insects. In the second step of our study, we analysed sequences of specific gene regions for a more detailed phylogeny of selected endosymbiont genera. As a result we identified the presence of <it>Rickettsia</it> and “<it>Candidatus</it> Nardonella<it>”</it> endosymbionts in <it>Otiorhynchus</it> spp.. This knowledge is an important step in exploring bacteria-insect associations for potential use in insect pest control.</p

Studie zum aktiven und passiven Wahlrecht von Menschen mit Behinderung

Die Studie beschäftigt sich mit den Voraussetzungen und Grenzen der in § 13 Nr. 2 und 3 Bundeswahlgesetz (BWG) geregelten Wahlrechtsausschlüsse. Sie betreffen Menschen, für die zur Besorgung aller ihrer Angelegenheiten ein Betreuer nicht nur durch einstweilige Anordnung bestellt ist (Nr.2) sowie Menschen, die sich auf Grund einer Anordnung nach § 63 in Verbindung mit § 20 des Strafgesetzbuches in einem psychiatrischen Krankenhaus befinden (Nr. 3). Die Untersuchung verfolgt einen interdisziplinären Ansatz. In ihrem sozialwissenschaftlichen Teil erfolgen empirische Erhebungen über die Anzahl der bundesweit von den genannten Wahlrechtsausschlüssen Betroffenen, deren soziodemographische Merkmale sowie der den Ausschlüssen zugrundeliegenden Krankheitsbildern. Außerdem werden im klinisch-psychologischen Teil die Betroffenenperspektive einbezogen, bestehende und mögliche Assistenzsysteme auch im internationalen Vergleich skizziert und der objektivierbare Bedarf nach Assistenz und die subjektiven Bedürfnisse hinsichtlich derartiger Unterstützungs-und Assistenzmaßnahmen dargestellt. In den juristischen Teilen erfolgt eine Untersuchung der Wahlrechtsausschlüsse in völkerrechtlicher und - damit in Wechselbeziehung stehend - verfassungsrechtlicher Perspektive. Im Mittelpunkt der völkerrechtlichen Untersuchung stehen die Vorgaben durch die Behindertenrechtskonvention (BRK). Deren Vorschriften werden nach den Regeln der Auslegung völkerrechtlicher Verträge interpretiert und im Kontext der Meinung des BRK-Ausschusses sowie der politischen Dynamik in der internationalen Gemeinschaft diskutiert. Im Zentrum des verfassungsrechtlichen Teils der Studie steht zunächst die verfassungsrechtliche Maßstabsentfaltung und hier namentlich das Zusammenspiel der verfassungsrechtlichen Wahlrechtsgrundsätze mit den spezifischen Benachteiligungsverboten von Menschen mit Behinderungen sowie der Bedeutung der BRK. § 13 Nr. 2 und 3 BWG werden sodann an diesem Maßstab überprüft. Abschließend zeigt die Studie 4 Handlungsoptionen auf.The study deals with the prerequisites and the limits of the disenfranchisement regulated in Section 3 No. 2 and 3 Federal Electoral Act. These provisions concern persons on whose behalf a custodian has been appointed to attend to all his or her affairs (No. 2) as well as persons who are accommodated in a psychiatric hospital under an order pursuant to Section 63 of the Penal Code in conjunction with Section 20 of the Penal Code. The study employs an interdisciplinary approach. In the sections of the social science analysis, an empirical survey concerning the number of persons affected in Germany by such disenfranchisement as mentioned above has been made, their sociodemographic attributes as well as the clinical situation under-pinning their disenfranchisement. In addition, the clinical-psychological section of the study describes the existing as well as alternative assistance systems employed internationally and illustrates the individual needs for assistance. The social-science sections are followed by two legal considerations, examining the current German disenfranchisement provisions from both an international and constitutional law perspective, the latter of which is mindful of international law. At the centre of the public international law section stands the International Convention on the Rights of Persons with Disabilities (CRPD). The provisions of this treaty are interpreted in accordance with the customary international rules of treaty interpretation and the result is discussed in the light of the opinion of the CRPD Committee and the political dynamics of the international community. The focus of the section dealing with the German Constitution is based on relevant constitutional standards and principles, especially the interplay between constitutional electoral principle with specific constitutional law on discrimination against people with disabilities as well as similar international law provisions. Section 13 No. 2 and 3 Federal Electoral Act is examined in the light of the constitutional benchmark established. Following on from a comparative law section, the study presents possible options for legislative action

What makes a young assertive bystander? The effect of intergroup contact, empathy, cultural openness, and in-Group bias on assertive bystander intervention intentions.

The present research tests the indirect effects of intergroup contact on adolescents’ bystander intervention intentions via four potential mediators: “empathy,” “cultural openness,” “in-group bias,” and “intergroup anxiety.” British adolescents (N = 855), aged 11–13 years, completed measures of intergroup (interethnic) contact and the identified indirect variables. Intended bystander behavior was measured by presenting participants with an intergroup (immigrant) name-calling scenario. Participants rated the extent to which they would behave assertively. The findings extend previous intergroup contact research by showing a significant indirect effect of intergroup contact on assertive bystander intentions via empathy, cultural openness and in-group bias (but not via intergroup anxiety). Theoretical implications and practical suggestions for future prejudice-reduction interventions are discussed

Metabolic fluxes for nutritional flexibility of Mycobacterium tuberculosis.

The co-catabolism of multiple host-derived carbon substrates is required by Mycobacterium tuberculosis (Mtb) to successfully sustain a tuberculosis infection. However, the metabolic plasticity of this pathogen and the complexity of the metabolic networks present a major obstacle in identifying those nodes most amenable to therapeutic interventions. It is therefore critical that we define the metabolic phenotypes of Mtb in different conditions. We applied metabolic flux analysis using stable isotopes and lipid fingerprinting to investigate the metabolic network of Mtb growing slowly in our steady-state chemostat system. We demonstrate that Mtb efficiently co-metabolises either cholesterol or glycerol, in combination with two-carbon generating substrates without any compartmentalisation of metabolism. We discovered that partitioning of flux between the TCA cycle and the glyoxylate shunt combined with a reversible methyl citrate cycle is the critical metabolic nodes which underlie the nutritional flexibility of Mtb. These findings provide novel insights into the metabolic architecture that affords adaptability of bacteria to divergent carbon substrates and expand our fundamental knowledge about the methyl citrate cycle and the glyoxylate shunt

Prejudice-reduction in culturally mixed classrooms : The development and assessment of a theory-driven prejudice-reduction intervention among majority and minority youth in Finland

Peer reviewe

Inter-rater reliability of the Full Outline of UnResponsiveness score and the Glasgow Coma Scale in critically ill patients : a prospective observational study

The Glasgow Coma Scale (GCS) is the most widely used scoring system for comatose patients in intensive care. Limitations of the GCS include the impossibility to assess the verbal score in intubated or aphasic patients, and an inconsistent inter-rater reliability. The FOUR (Full Outline of UnResponsiveness) score, a new coma scale not reliant on verbal response, was recently proposed. The aim of the present study was to compare the inter-rater reliability of the GCS and the FOUR score among unselected patients in general critical care. A further aim was to compare the inter-rater reliability of neurologists with that of intensive care unit (ICU) staff

Leaf microbiota of strawberries as affected by biological control agents

The increasing use of biological control agents (BCAs) against Botrytis cinerea in strawberry raises the question of whether there are any undesirable impacts of foliar applications of BCAs on nontarget microorganisms in the phyllosphere. Therefore, our objective was to investigate this issue within a field study. Strawberry plants were repeatedly sprayed with three BCAs—namely, RhizoVital 42 fl. (Bacillus amyloliquefaciens FZB42), Trianum-P (Trichoderma harzianum T22), and Naturalis (Beauveria bassiana ATCC 74040)—to suppress Botrytis cinerea infections. Microbial communities of differentially treated leaves were analyzed using plate counts and pyrosequencing and compared with the microbial community of nontreated leaves. Plate count results indicate that the applied Bacillus and Trichoderma spp. survived in the strawberry phyllosphere throughout the strawberry season. However, no significant impacts on the leaf microbiota could be detected by this culture-dependent technique. Pyrosequencing of internal transcribed spacer ribosomal RNA and 16S RNA sequences revealed a change in fungal composition and diversity at class level after the introduction of T. harzianum T22 to the phyllosphere, whereas the bacterial composition and diversity was not affected by either this Trichoderma preparation or the other two BCAs. Our results suggest that pyrosequencing represents a useful method for studying microbial interactions in the phyllosphere

Congruent geographic variation in saccular otolith shape across multiple species of African cichlids

The otoliths of teleost fishes exhibit a great deal of inter- and intra-species shape variation. The ecomorphology of the saccular otolith is often studied by comparing its shape across species and populations inhabiting a range of environments. However, formal tests are often lacking to examine how closely variation in otolith shape follows the genetic drift of a neutral trait. Here, we examine patterns of saccular otolith shape variation in four species of African cichlid fishes, each sampled from three field sites. All four species showed the greatest level of otolith shape variation along two principal component axes, one pertaining to otolith height and another to the prominence of an anterior notch. Fish collected from the same site possessed similarities in saccular otolith shape relative to fish from other sites, and these 'site-difference' signatures were consistent across species and observable in both sexes. Sex-differences in saccular otolith shape differed in magnitude from site to site. Population differences in saccular otolith shape did not covary with neutral genetic differentiation between those populations. Otolith height, in particular, displayed large site similarities across species, weak correlation with neutral genetic variation, and strong sex differences, collectively suggesting that otolith shape represents a selectively non-neutral trait.publishe

Communities of Niche-optimized Strains (CoNoS) – Design and creation of stable, genome-reduced co-cultures

Current bioprocesses for production of value-added compounds are mainly based on pure cultures that are composed of rationally engineered strains of model organisms with versatile metabolic capacities. However, in the comparably well-defined environment of a bioreactor, metabolic flexibility provided by various highly abundant biosynthetic enzymes is much less required and results in suboptimal use of carbon and energy sources for compound production. In nature, non-model organisms have frequently evolved in communities where genome-reduced, auxotrophic strains cross-feed each other, suggesting that there must be a significant advantage compared to growth without cooperation. To prove this, we started to create and study synthetic communities of niche-optimized strains (CoNoS) that consists of two strains of the same species Corynebacterium glutamicum that are mutually dependent on one amino acid. We used both the wild-type and the genome-reduced C1* chassis for introducing selected amino acid auxotrophies, each based on complete deletion of all required biosynthetic genes. The best candidate strains were used to establish several stably growing CoNoS that were further characterized and optimized by metabolic modelling, microfluidic experiments and rational metabolic engineering to improve amino acid production and exchange. Finally, the engineered CoNoS consisting of an l-leucine and l-arginine auxotroph showed a specific growth rate equivalent to 83% of the wild type in monoculture, making it the fastest co-culture of two auxotrophic C. glutamicum strains to date. Overall, our results are a first promising step towards establishing improved biobased production of value-added compounds using the CoNoS approach