Soil respiration is stimulated by elevated CO2 and reduced by summer drought: three years of measurements in a multifactor ecosystem manipulation experiment in a temperate heathland (CLIMAITE)

This study investigated the impact of predicted future climatic and atmospheric conditions on soil respiration (R_S) in a Danish Calluna‐Deschampsia‐heathland. A fully factorial in situ experiment with treatments of elevated atmospheric CO₂ (+130 ppm), raised soil temperature (+0.4 °C) and extended summer drought (5–8% precipitation exclusion) was established in 2005. The average R_S, observed in the control over 3 years of measurements (1.7 μmol CO₂ m^?2 sec^?1), increased 38% under elevated CO₂, irrespective of combination with the drought or temperature treatments. In contrast, extended summer drought decreased R_S by 14%, while elevated soil temperature did not affect R_S overall. A significant interaction between elevated temperature and drought resulted in further reduction of R_S when these treatments were combined. A detailed analysis of short‐term R_S dynamics associated with drought periods showed that R_S was reduced by ~50% and was strongly correlated with soil moisture during these events. Recovery of R_S to pre‐drought levels occurred within 2 weeks of rewetting; however, unexpected drought effects were observed several months after summer drought treatment in 2 of the 3 years, possibly due to reduced plant growth or changes in soil water holding capacity. An empirical model that predicts R_S from soil temperature, soil moisture and plant biomass was developed and accounted for 55% of the observed variability in R_S. The model predicted annual sums of R_S in 2006 and 2007, in the control, were 672 and 719 g C m^?2 y^?1, respectively. For the full treatment combination, i.e. the future climate scenario, the model predicted that soil respiratory C losses would increase by ~21% (140–150 g C m^?2 y^?1). Therefore, in the future climate, stimulation of C storage in plant biomass and litter must be in excess of 21% for this ecosystem to not suffer a reduction in net ecosystem exchange.     

Detection of a quantitative trait locus associated with resistance to Ascaris suum infection in pigs

Helminths almost invariably have an over-dispersed distribution in the host population. Human and animal studies have provided evidence suggesting that a large part of this variation is due to host genetic factors. Recently, the heritability for roundworm (Ascaris suum) infection levels in pigs was estimated to be 0.45. We used single nucleotide polymorphism markers to perform a whole-genome scan on 195 pigs experimentally infected with A. suum. A putative quantitative trait locus for worm burden on chromosome 4 covering 2.5 Mbp was identified by measured genotype analysis, although none of the SNPs reached genome-wide significance. To validate the putative quantitative trait locus, we genotyped two of the SNPs within the region in unrelated, informative animals exposed to experimental or natural infections and from which we had worm counts and/or faecal egg counts; the validation studies showed that one of the SNPs (TXNIP) was associated with total worm burden (P < 0.001) and adult worm burden(P < 0.0001), whereas the other SNP (ARNT) was associated with adult worm burden (P < 0.025) in these populations. We were thus able to confirm the existence of the quantitative trait locus on chromosome 4.This is to our knowledge the first report of a quantitative trait locus associated with helminth burden in pigs.     

Delineation of the Species Haemophilus influenzae by Phenotype,Multilocus Sequence Phylogeny,and Detection of Marker Genes

To obtain more information on the much-debated definition of prokaryotic species, we investigated the borders of Haemophilus influenzae by comparative analysis of H. influenzae reference strains with closely related bacteria including strains assigned to Haemophilus haemolyticus, cryptic genospecies biotype IV, and the never formally validated species “Haemophilus intermedius”. Multilocus sequence phylogeny based on six housekeeping genes separated a cluster encompassing the type and the reference strains of H. influenzae from 31 more distantly related strains. Comparison of 16S rRNA gene sequences supported this delineation but was obscured by a conspicuously high number of polymorphic sites in many of the strains that did not belong to the core group of H. influenzae strains. The division was corroborated by the differential presence of genes encoding H. influenzae adhesion and penetration protein, fuculokinase, and Cu,Zn-superoxide dismutase, whereas immunoglobulin A1 protease activity or the presence of the iga gene was of limited discriminatory value. The existence of porphyrin-synthesizing strains (“H. intermedius”) closely related to H. influenzae was confirmed. Several chromosomally encoded hemin biosynthesis genes were identified, and sequence analysis showed these genes to represent an ancestral genotype rather than recent transfers from, e.g., Haemophilus parainfluenzae. Strains previously assigned to H. haemolyticus formed several separate lineages within a distinct but deeply branching cluster, intermingled with strains of “H. intermedius” and cryptic genospecies biotype IV. Although H. influenzae is phenotypically more homogenous than some other Haemophilus species, the genetic diversity and multicluster structure of strains traditionally associated with H. influenzae make it difficult to define the natural borders of that species.Bacterial diversity is usually expected to be organized into phenotypic and genetic clusters recognized as species. The diversity within species is thought to be confined by natural forces, although uncertainty exists as to the nature of the cohesion that constrains this diversity. There is increasing evidence that recombination occurs between distantly related bacteria and that barriers to this process, which could be used to define species naturally, are not always apparent (7, 11). As the bacterial species concept is central to taxonomy and evolutionary biology and has consequences for other disciplines such as the discrimination of pathogens from related bacteria with no pathogenic potential, there is a need to reevaluate to what extent accepted species are indeed coherent units.The gram-negative, nonmotile, facultatively anaerobic bacterium Haemophilus influenzae is a commensal organism of the pharynx frequently involved in localized infections of the respiratory tract, middle ear, and conjunctiva and sometimes in invasive infections such as meningitis or bacteremia. Presumptive identification is based on inability to synthesize NAD (V factor dependence) and porphyrin/hemin (X factor dependence). The latter characteristic is particularly notable as it is shared with few other microorganisms. Further metabolic characteristics of H. influenzae include fermentation of d-xylose, d-ribose, and d-galactose but not sucrose or d-mannose (17). By DNA analysis the X-factor-dependent species H. influenzae, Haemophilus aegyptius, and Haemophilus haemolyticus constitute a closely related group (4, 5, 14, 35), and housekeeping gene nucleotide dissimilarities between these species are exceeded by intraspecific nucleotide dissimilarities within the commensal species Haemophilus parainfluenzae (33).H. aegyptius was conceived as a species distinct from H. influenzae with a particular propensity to cause conjunctivitis (37). However, the existence of H. aegyptius as a separate species is controversial (5), and the designation H. influenzae biogroup aegyptius has been proposed to include traditional H. aegyptius and the closely related clones responsible for the invasive infection known as Brazilian purpuric fever (3), although formally the epithet aegyptius has priority. H. haemolyticus is distinguished from H. influenzae primarily by its hemolytic action on erythrocytes. A recent characterization of a large number of presumptive H. influenzae isolates from the respiratory tract of patients with chronic obstructive airway disease demonstrated that 102 of 258 strains clustered with the type strain of H. haemolyticus by 16S rRNA gene sequence, although many of the isolates were nonhemolytic. In contrast to H. influenzae, the atypical isolates were not associated with exacerbations of the disease (31), emphasizing the need to discriminate between the two species.Genetic methods have identified unnamed taxa of Haemophilus related to H. influenzae. Variant strains isolated from the genitourinary tract were first reported from Canada (1) and have been further studied by Quentin and coworkers (13, 39). These “cryptic genospecies biotype IV” strains (negative for tryptophanase/indole production and positive for urease and ornithine decarboxylase) are distinguishable from typical strains of H. influenzae by multilocus enzyme electrophoresis, 16S rRNA gene sequence, and DNA hybridization. However, only some biotype IV strains belong to the cryptic genospecies, and the lack of phenotypic characters enabling discrimination from H. influenzae has been an obstacle to the formal validation as a species. Another taxon, “Haemophilus intermedius,” was suggested (but never validly published) by Burbach (4). By DNA hybridization and by selected phenotypic traits, these strains take up an intermediate position between H. influenzae and H. parainfluenzae. Two subspecies were described, “Haemophilus intermedius subsp. intermedius,” capable of synthesizing porphyrin from δ-aminolevulinic acid and of fermenting sucrose, and “Haemophilus intermedius subsp. gazogenes,” characterized by the fermentation of mannose and the production of gas from glucose.A number of investigations have addressed the population structure of H. influenzae with assessment of particular virulence traits associated with specific subgroups or clades (9, 26, 32, 41). In the present study we attempted to define the borders of the species by performing phenotypic characterization, 16S rRNA sequence comparison, multilocus housekeeping gene sequence phylogeny, and detection of putative virulence and marker genes on a collection of 42 strains of H. influenzae and representatives of the above-mentioned closely related bacteria.     

Life cycle assessment (LCA) of industrial milk production

A Life Cycle Assessment (LCA) was carried out for milk production extending from the origin of the inputs to the agricultural step to the consumer phase and the waste management of the packaging. Three Norwegian dairies of different sizes and degree of automation were studied. The main objectives were to find any hot spots in the life cycle of milk, to determine the significance of the dairy size and degree of automation, and to study the influence of transport. The agriculture was found to be the main hot spot for almost all the environmental themes studied, although the dairy processing, packaging, consumer phase and waste management were also of importance. The consumer phase was the main contributor to photo-oxidant formation and important regarding eutrophication. The small dairy was found to have a greater environmental impact than the middle-sized and the largest dairies. The transport did not have any major influence.     

Balance between matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in the cervical mucus plug estimated by determination of free non-complexed TIMP

Background  

The cervical mucus plug (CMP) is a semi-solid structure with antibacterial properties positioned in the cervical canal during pregnancy. The CMP contains high concentrations of matrix metalloproteinase 8 and 9 (MMP-8, MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1). This indicates a potential to degrade extracellular matrix components depending on the balance between free non-complexed inhibitors and active enzymes.     

Chemical modification patterns compatible with high potency dicer-substrate small interfering RNAs

Dicer-substrate small interfering RNAs (DsiRNAs) are synthetic RNA duplexes that are processed by Dicer into 21-mer species and show improved potency as triggers of RNA interference, particularly when used at low dose. Chemical modification patterns that are compatible with high potency 21-mer small interfering RNAs have been reported by several groups. However, modification patterns have not been studied for Dicer-substrate duplexes. We therefore synthesized a series of chemically modified 27-mer DsiRNAs and correlated modification patterns with functional potency. Some modification patterns profoundly reduced function although other patterns maintained high potency. Effects of sequence context were observed, where the relative potency of modification patterns varied between sites. A modification pattern involving alternating 2'-O-methyl RNA bases was developed that generally retains high potency when tested in different sites in different genes, evades activation of the innate immune system, and improves stability in serum.     

A form of albinism in cattle is caused by a tyrosinase frameshift mutation

We used PCR amplification of cDNA prepared from skin biopsies to determine the full-length protein-coding sequence of tyrosinase (TYR) in cattle of several coat colors. An insertion of a cytosine was detected in an albino Braunvieh calf, which resulted in a frameshift which caused a premature stop codon at residue 316. This insertion was found in the homozygous state in this calf and the genomic DNA of two related albino calves. All six parents of these calves were heterozygous for this insertion. However, an albino Holstein calf did not have this insertion, nor was any other mutation detected in the partial TYR sequence obtained from the genomic DNA available. Diagnostic genotyping tests were developed to detect this mutation in Braunvieh cattle.     

The GENETPIG database: a tool for comparative mapping in pig (Sus scrofa)

The GENETPIG database has been established for storing and disseminating the results of the European project: 'GENETPIG: identification of genes controlling economic traits in pig'. The partners of this project have mapped about 630 porcine and human ESTs onto the pig genome. The database collects the mapping results and links them to other sources of mapping data; this includes pig maps as well as available comparative mapping information. Functional annotation of the mapped ESTs is also given when a significant similarity to cognate genes was established. The database is accessible for consultation via the Internet at http://www.infobiogen.fr/services/Genetpig/.