Northern glacial refugia for the pygmy shrew Sorex minutus in Europe revealed by phylogeographic analyses and species distribution modelling

The southern European peninsulas (Iberian, Italian and Balkan) are traditionally recognized as glacial refugia from where many species colonized central and northern Europe after the Last Glacial Maximum (LGM). However, evidence that some species had more northerly refugia is accumulating from phylogeographic, palaeontological and palynological studies, and more recently from species distribution modelling (SDM), but further studies are needed to test the idea of northern refugia in Europe. Here, we take a rarely implemented multidisciplinary approach to assess if the pygmy shrew Sorex minutus, a widespread Eurasian mammal species, had northern refugia during the LGM, and if these influenced its postglacial geographic distribution. First, we evaluated the phylogeographic and population expansion patterns using mtDNA sequence data from 123 pygmy shrews. Then, we used SDM to predict present and past (LGM) potential distributions using two different training data sets, two different algorithms (Maxent and GARP) and climate reconstructions for the LGM with two different general circulation models. An LGM distribution in the southern peninsulas was predicted by the SDM approaches, in line with the occurrence of lineages of S. minutus in these areas. The phylogeographic analyses also indicated a widespread and strictly northern‐central European lineage, not derived from southern peninsulas, and with a postglacial population expansion signature. This was consistent with the SDM predictions of suitable LGM conditions for S. minutus occurring across central and eastern Europe, from unglaciated parts of the British Isles to much of the eastern European Plain. Hence, S. minutus likely persisted in parts of central and eastern Europe during the LGM, from where it colonized other northern areas during the late‐glacial and postglacial periods. Our results provide new insights into the glacial and postglacial colonization history of the European mammal fauna, notably supporting glacial refugia further north than traditionally recognized.     

Characterisation of social impacts in LCA. Part 2: implementation in six company case studies

Background, aim and scope

A characterisation model based on multi-criteria indicators has been developed for each of four impact categories representing the labour rights according to the conventions of the International Labour Organisation (ILO) covering: forced labour, discrimination, restrictions of freedom of association and collective bargaining and child labour (Dreyer et al., Int J Life Cycle Assess, 2010a, in press). These impact categories are considered by the authors to be among the obligatory impact categories in a Social LCA. The characterisation models combine information about the way a company manages its behaviour towards some of its important stakeholders, its employees, with information about the geographical location and branch of industry of the company and the risk of violations of these workers' rights inherent in the setting of the company. The result is an indicator score which for each impact category represents the risk that violations occur in the company. In order to test the feasibility and relevance of the developed methodology, it is tested on real cases.

Materials and methods

The developed characterisation models are applied to six cases representing individual manufacturing companies from three different continents. Five of the case companies are manufacturing companies while the sixth is a knowledge company. The application involves scoring the management efforts of the case company in a multi-criteria scorecard and translating the scores into an aggregated performance score, which represents the effort of the management in order to prevent violations of the workers' rights to occur in the company. The company performance score is multiplied by a contextual adjustment score which reflects the risk of violations taking place in the context (in terms of geographical location or industrial branch or sector) of the company. The resulting indicator score represents the risk that violations take place of the labour right represented by the impact category.

Results

The social impact characterisation is performed for each of the six case studies using the methodology earlier developed. The procedure and outcome are documented through all the intermediary results shown for all four obligatory impact categories for each of the six case studies.

Discussion

The results are judged against the risk which was observed during visits and interviews at each of the six case companies, and their realism and relevance are discussed. They are found to be satisfactory for all four impact categories for the manufacturing companies, but there are some problems for two of the impact categories in the case company which represents knowledge work, and it is discussed how these problems may be addressed through change of the underlying scorecard or the way in which the scoring is translated into a company performance score.

Conclusions

It is concluded that it is feasible to perform a characterisation of the impacts related to the four obligatory impact categories representing the labour rights according to the conventions of the ILO covering: forced labour, discrimination, restrictions of freedom of association and collective bargaining and child labour. When compared with the observed situation in the companies, the results are also found to be relevant and realistic.

Recommendations and perspectives

The proposed characterisation method is rather time-consuming and cannot realistically be applied to all companies in the product system. It must therefore be combined with less time-requiring screening methods which can help identify the key companies in the life cycle for which a detailed analysis is required. The possibility to apply country- or industry sector-based information is discussed, and while it is found useful to identify low-risk companies and eliminate them from more detailed studies, the ability of the screening methods to discriminate between companies located in medium and high-risk contexts is questionable.     

Searching for Mesophilic Thermotogales Bacteria: “Mesotogas” in the Wild

All cultivated Thermotogales are thermophiles or hyperthermophiles. However, optimized 16S rRNA primers successfully amplified Thermotogales sequences from temperate hydrocarbon-impacted sites, mesothermic oil reservoirs, and enrichment cultures incubated at <46°C. We conclude that distinct Thermotogales lineages commonly inhabit low-temperature environments but may be underreported, likely due to “universal” 16S rRNA gene primer bias.Thermotogales, a bacterial group in which all cultivated members are anaerobic thermophiles or hyperthermophiles (5), are rarely detected in anoxic mesothermic environments, yet their presence in corresponding enrichment cultures, bioreactors, and fermentors has been observed using metagenomic methods and 16S rRNA gene amplification (6) (see Table S1 in the supplemental material). The most commonly detected lineage is informally designated here “mesotoga M1” (see Table S1 in the supplemental material). PCR experiments indicated that mesotoga M1 sequences amplified inconsistently using “universal” 16S rRNA gene primers, perhaps explaining their poor detection in DNA isolated from environmental samples (see text and Table S2 in the supplemental material). We therefore designed three 16S rRNA PCR primer sets (Table ​(Table1)1) targeting mesotoga M1 bacteria and their closest cultivated relative, Kosmotoga olearia. Primer set A was the most successful set, detecting a wider diversity of Thermotogales sequences than set B and being more Thermotogales-specific than primer set C (Table ​(Table22).

TABLE 1.

Primers targeting mesotoga M1 bacteria constructed and used in this study

Investigation of Chlamydiaceae in semen and cauda epididymidis and seroprevalence of Chlamydophila abortus in breeding bulls

Background

Reproductive disorders associated with chlamydial infection have been reported worldwide in cattle and there are indications of potential venereal transmission.

Methods

Semen samples from 21 dairy bulls and cauda epididymidis tissue samples from 43 beef bulls were analysed for chlamydial agent by real-time polymerase chain reaction (PCR) including an internal amplification control (mimic). Additionally, presence of antibodies against Chlamydophila (Cp.) abortus among the bulls was investigated with the commercial Pourquier^® ELISA Cp. abortus serum verification kit.

Results

No chlamydial agent was detected by PCR in either the semen samples or in the tissue samples. Additionally, no antibodies against Cp. abortus were detected.

Conclusions

The results suggest that Cp. abortus is very rare, or absent in Swedish bulls and thus the risk for venereal transmission of chlamydial infection through their semen is low. However, because Chlamydophila spp. infection rates seem to differ throughout the world, it is essential to clarify the relative importance of transmission of the infection through semen on cattle fertility.     

Confidence from uncertainty - A multi-target drug screening method from robust control theory

Background  

Robustness is a recognized feature of biological systems that evolved as a defence to environmental variability. Complex diseases such as diabetes, cancer, bacterial and viral infections, exploit the same mechanisms that allow for robust behaviour in healthy conditions to ensure their own continuance. Single drug therapies, while generally potent regulators of their specific protein/gene targets, often fail to counter the robustness of the disease in question. Multi-drug therapies offer a powerful means to restore disrupted biological networks, by targeting the subsystem of interest while preventing the diseased network from reconciling through available, redundant mechanisms. Modelling techniques are needed to manage the high number of combinatorial possibilities arising in multi-drug therapeutic design, and identify synergistic targets that are robust to system uncertainty.     

Structure and Mode-of-Action of the Two-Peptide (Class-IIb) Bacteriocins

This review focuses on the structure and mode-of-action of the two-peptide (class-IIb) bacteriocins that consist of two different peptides whose genes are next to each other in the same operon. Optimal antibacterial activity requires the presence of both peptides in about equal amounts. The two peptides are synthesized as preforms that contain a 15–30 residue double-glycine-type N-terminal leader sequence that is cleaved off at the C-terminal side of two glycine residues by a dedicated ABC-transporter that concomitantly transfers the bacteriocin peptides across cell membranes. Two-peptide bacteriocins render the membrane of sensitive bacteria permeable to a selected group of ions, indicating that the bacteriocins form or induce the formation of pores that display specificity with respect to the transport of molecules. Based on structure–function studies, it has been proposed that the two peptides of two-peptide bacteriocins form a membrane-penetrating helix–helix structure involving helix–helix-interacting GxxxG-motifs that are present in all characterized two-peptide bacteriocins. It has also been suggested that the membrane-penetrating helix–helix structure interacts with an integrated membrane protein, thereby triggering a conformational alteration in the protein, which in turn causes membrane-leakage. This proposed mode-of-action is similar to the mode-of-action of the pediocin-like (class-IIa) bacteriocins and lactococcin A (a class-IId bacteriocin), which bind to a membrane-embedded part of the mannose phosphotransferase permease in a manner that causes membrane-leakage and cell death.     

Microbial responses to P addition in six South African forest soils

Forests growing on highly weathered soils are often phosphorus (P) limited and competition between geochemical and biological sinks affects their soil P dynamics. In an attempt to elucidate the factors controlling the relative importance of these two sinks, we investigated the relationship of between soil microbial growth kinetics and soil chemical properties following amendments with C, N and P in six South African forest soils. Microbial growth kinetics were determined from respiration curves derived from measurements of CO₂ effluxes from soil samples in laboratory incubations. We found that microbial growth rates after C + N additions were positively related to NaOH-extractable P and decreased with soil depth, whereas the lag time (the time between substrate addition and exponential growth) was negatively related to extractable P. However, the growth rate and lag time were unrelated to the soil’s sorption properties or Al and Fe contents. Our results indicate that at least some of the NaOH-extractable inorganic P may be biologically available within a relatively short time (days to weeks) and might be more labile than previously thought. Our results also show that microbial utilization of C + N only seemed to be constrained by P in the deeper part of the soil profiles.     

Enhanced oligonucleotide-nanoparticle conjugate stability using thioctic acid modified oligonucleotides

Metallic nanoparticles of gold functionalized with oligonucleotides conventionally use a terminal thiol modification and have been used in a wide range of applications. Although readily available, the oligonucleotide–nanoparticle conjugates prepared in this way suffer from a lack of stability when exposed to a variety of small molecules or elevated temperatures. If silver is used in place of gold then this lack of stability is even more pronounced. In this study we report the synthesis of highly stabilized oligonucleotide–nanoparticle conjugates using a simple oligonucleotide modification. A modified solid support was used to generate 3′-thioctic acid modified oligonucleotides by treatment with an N-hydroxysuccimidyl ester of thioctic acid. Unusually, both gold and silver nanoparticles have been investigated in this study and show that these disulphide-modified oligonucleotide probes offer significant improvements in nanoparticle stability when treated with dithiothreitol (DTT) compared with monothiol analogues. This is a significant advance in oligonucleotide–nanoparticle conjugate stability and for the first time allows silver nanoparticles to be prepared that are more stable than standard gold-thiol functionalized nanoparticles. This opens up the possibility of using silver nanoparticles functionalized with oligonucleotides as an alternative to gold.