Specific and sensitive detection of Nosema bombi (Microsporidia : Nosematidae) in bumble bees (Bombus spp.; Hymenoptera : Apidae) by PCR of partial rRNA gene sequences

A polymerase chain reaction (PCR) based method was developed for the specific and sensitive diagnosis of the microsporidian parasite Nosema bombi in bumble bees (Bombus spp.). Four primer pairs, amplifying ribosomal RNA (rRNA) gene fragments, were tested on N. bombi and the related microsporidia Nosema apis and Nosema ceranae, both of which infect honey bees. Only primer pair Nbombi-SSU-Jf1/Jr1 could distinguish N. bombi (323 bp amplicon) from these other bee parasites. Primer pairs Nbombi-SSU-Jf1/Jr1 and ITS-f2/r2 were then tested for their sensitivity with N. bombi spore concentrations from 107 down to 10 spores diluted in 100 mu l of either (i) water or (ii) host bumble bee homogenate to simulate natural N. bombi infection (equivalent to the DNA from 10(6) spores down to 1 spore per PCR). Though the N. bombi-specific primer pair Nbombi-SSU-Jf1/Jr1 was relatively insensitive, as few as 10 spores per extract (equivalent to 1 spore per PCR) were detectable using the N. bombi-non-specific primer pair ITS-f2/r2, which amplifies a short fragment of similar to 120 bp. Testing 99 bumble bees for N. bombi infection by light microscopy versus PCR diagnosis with the highly sensitive primer pair ITS-f2/r2 showed the latter to b more accurate. PCR diagnosis of N. bombi using a combination of two primer pairs (Nbombi-SSU-Jf1/Jr1 and ITS-f2/r2) provides increased specificity, sensitivity, and detection of all developmental stages compared with light microscopy. (c) 2005 Elsevier Inc. All rights reserved.

Species boundaries and phylogenetic relationships within the green algal genus Codium (Bryopsidales) based on plastid DNA sequences

Despite the potential model role of the green algal genus Codium for studies of marine speciation and evolution, there have been difficulties with species delimitation and a molecular phylogenetic framework was lacking. In the present study, 74 evolutionarily significant units (ESUs) are delimited using 227 rbcL exon 1 sequences obtained from specimens collected throughout the genus' range. Several morpho-species were shown to be poorly defined, with some clearly in need of lumping and others containing pseudo-cryptic diversity. A phylogenetic hypothesis of 72 Codium ESUs is inferred from rbcL exon 1 and rps3-rp/16 sequence data using a conventional nucleotide substitution model (GTR + Gamma + I), a codon position model and a covariotide (covarion) model, and the fit of a multitude of substitution models and alignment partitioning strategies to the sequence data is reported. Molecular clock tree rooting was carried out because out-group rooting was probably affected by phylogenetic bias. Several aspects of the evolution of morphological features of Codium are discussed and the inferred phylogenetic hypothesis is used as a framework to study the biogeography of the genus, both at a global scale and within the Indian Ocean. (c) 2007 Elsevier Inc. All rights reserved.

Searching and planning: Young children's reasoning about past and future event sequences

Six experiments examined children's ability to make inferences using temporal order information. Children completed versions of a task involving a toy zoo; one version required reasoning about past events (search task) and the other required reasoning about future events (planning task). Children younger than 5 years failed both the search and the planning tasks, whereas 5-year-olds passed both (Experiments 1 and 2). However, when the number of events in the sequence was reduced (Experiment 3), 4-year-olds were successful on the search task but not the planning task. Planning difficulties persisted even when relevant cues were provided (Experiments 4 and 5). Experiment 6 showed that improved performance on the search task found in Experiment 3 was not due to the removal of response ambiguity.

Methods and code for 'classical' age-modelling of radiocarbon sequences.

Age–depth models form the backbone of most palaeoenvironmental studies. However, procedures for constructing chronologies vary between studies, they are usually not explained sufficiently, and some are inadequate for handling calibrated radiocarbon dates. An alternative method based on importance sampling through calibrated dates is proposed. Dedicated R code is presented which works with calibrated radiocarbon as well as other dates, and provides a simple, systematic, transparent, documented and customizable alternative. The code automatically produces age–depth models, enabling exploration of the impacts of different assumptions (e.g., model type, hiatuses, age offsets, outliers, and extrapolation).

Revised age estimate of the Mjauvotn tephra A on the Faroe Islands based on Bayesian modelling of C-14 dates from two lake sequences

Tephra horizons are potentially perfect time markers for dating and cross-correlation among diverse Holocene palaeoenvironmental records such as ice cores and marine and terrestrial sequences, but we need to trust their age. Here we present a new age estimate of the Holocene Mjauvotn tephra A using accelerator mass spectrometry C-14 dates from two lakes on the Faroe Islands. With Bayesian age modelling it is dated to 6668-6533 cal. a BP (68.2% confidence interval) - significantly older and better constrained than the previous age. Copyright (C) 2010 John Wiley & Sons, Ltd.

Analysis of transduction in wastewater bacterial populations by targeting the phage-derived 16S rRNA gene sequences

Bacterial 16S rRNA genes transduced by bacteriophages were identified and analyzed in order to estimate the extent of the bacteriophage-mediated horizontal gene transfer in the wastewater environment. For this purpose, phage and bacterial DNA was isolated from the oxidation tank of a municipal wastewater treatment plant. Phylogenetic analysis of the 16S rRNA gene sequences cloned from a phage metagenome revealed that bacteriophages transduce genetic material in several major groups of bacteria. The groups identified were as follows: Betaproteobacteria, Gammaproteobacteria, Alphaproteobacteria, Actinomycetales and Firmicutes. Analysis of the 16S rRNA gene sequences in the total bacterial DNA from the same sample revealed that several bacterial groups found in the oxidation tank were not present in the phage metagenome (e.g. Deltaproteobacteria, Nitrospira, Planctomycetes and many Actinobacteria genera). These results suggest that transduction in a wastewater environment occurs in several bacterial groups; however, not all species are equally involved into this process. The data also showed that a number of distinctive bacterial strains participate in transduction-mediated gene transfer within identified bacterial groupings. Denaturing gradient gel electrophoresis analysis confirmed that profiles of the transduced 16S rRNA gene sequences and those present in the whole microbial community show significant differences.

Extent and variation of phage-borne bacterial 16S rRNA gene sequences in wastewater environments

Phage metagenomes isolated from wastewater over a 12-month period were analyzed. The results suggested that various strains of Proteobacteria, Bacteroidetes, and other phyla are likely to participate in transduction. The patterns of 16S rRNA sequences found in phage metagenomes did not follow changes in the total bacterial community.

Complete Genome Sequences of Three Propionibacterium acnes Isolates from the Type IA(2) Cluster

Propionibacterium acnes is an anaerobic Gram-positive bacterium that has been linked to a wide range of opportunistic human infections and conditions, most notably acne vulgaris (I. Kurokawa et al., Exp. Dermatol. 18:821-832, 2009). We now present the whole-genome sequences of three P. acnes strains from the type IA(2) cluster which were recovered from ophthalmic infections (A. McDowell et al., Microbiology 157:1990-2003, 2011).

A spatio-temporal 2D-models framework for human pose recovery in monocular sequences

This paper addresses the pose recovery problem of a particular articulated object: the human body. In this model-based approach, the 2D-shape is associated to the corresponding stick figure allowing the joint segmentation and pose recovery of the subject observed in the scene. The main disadvantage of 2D-models is their restriction to the viewpoint. To cope with this limitation, local spatio-temporal 2D-models corresponding to many views of the same sequences are trained, concatenated and sorted in a global framework. Temporal and spatial constraints are then considered to build the probabilistic transition matrix (PTM) that gives a frame to frame estimation of the most probable local models to use during the fitting procedure, thus limiting the feature space. This approach takes advantage of 3D information avoiding the use of a complex 3D human model. The experiments carried out on both indoor and outdoor sequences have demonstrated the ability of this approach to adequately segment pedestrians and estimate their poses independently of the direction of motion during the sequence. (c) 2008 Elsevier Ltd. All rights reserved.

REMA: A computer-based mapping tool for analysis of restriction sites in multiple DNA sequences

REMA is an interactive web-based program which predicts endonuclease cut sites in DNA sequences. It analyses Multiple sequences simultaneously and predicts the number and size of fragments as well as provides restriction maps. The users can select single or paired combinations of all commercially available enzymes. Additionally, REMA permits prediction of multiple sequence terminal fragment sizes and suggests suitable restriction enzymes for maximally discriminatory results. REMA is an easy to use, web based program which will have a wide application in molecular biology research. Availability: REMA is written in Perl and is freely available for non-commercial use. Detailed information on installation can be obtained from Jan Szubert (jan.szubert@gmail.com) and the web based application is accessible on the internet at the URL http://www.macaulay.ac.uk/rema. Contact: b.singh@macaulay.ac.uk. (C) 2007 Elsevier B.V. All rights reserved.