Presence and genetic variability of Piscine orthoreovirus genotype 1 (PRV‐1) in wild salmonids in Northern Europe and North Atlantic Ocean

Piscine orthoreovirus genotype 1 (PRV‐1) is widespread in farmed Atlantic salmon (Salmo salar L.) populations in northern Europe, Canada and Chile. PRV‐1 occurs in wild fish in Norway and Canada; however, little information of its geographical distribution in wild populations is currently available, and the effect of PRV‐1 infection in wild populations is currently unknown. In this study, we present the findings of a survey conducted on 1,130 wild salmonids sampled in Denmark, Sweden, Ireland, Faroe Islands, France, Belgium and Greenland between 2008 and 2017. PRV‐1 is reported for the first time in wild salmonids in Denmark, Sweden, Faroe Island and Ireland. The annual PRV‐1 prevalence ranged from 0% in France, Belgium and Greenland to 43% in Faroe Islands. In total, 66 samples tested positive for PRV‐1, including Atlantic salmon broodfish returning to spawn and Atlantic salmon collected at the feeding ground north of Faroe Islands. The phylogenetic analysis of S1 sequences of the PRV‐1 isolates obtained in this survey did not show systematic geographical distribution. This study sheds light on the spread and genetic diversity of the virus identified in populations of free‐living fish and provides rationale for screening wild broodfish used in restocking programmes.     

Dung application increases CH<Subscript>4</Subscript> production potential and alters the composition and abundance of methanogen community in restored peatland soils from Europe

Peatland restoration via rewetting aims to recover biological communities and biogeochemical processes typical to pristine peatlands. While rewetting promotes recovery of C accumulation favorable for climate mitigation, it also promotes methane (CH₄) emissions. The potential for exceptionally high emissions after rewetting has been measured for Central European peatland sites previously grazed by cattle. We addressed the hypothesis that these exceptionally high CH₄ emissions result from the previous land use. We analyzed the effects of cattle dung application to peat soils in a short- (2 weeks), a medium- (1 year) and a long-term (grazing) approach. We measured the CH₄ production potentials, determined the numbers of methanogens by mcrA qPCR, and analyzed the methanogen community by mcrA T-RFLP-cloning-sequencing. Dung application significantly increased the CH₄ production potential in the short- and the medium-term approach and non-significantly at the cattle-grazed site. The number of methanogens correlated with the CH₄ production in the short- and the long-term approach. At all three time horizons, we found a shift in methanogen community due to dung application and a transfer of rumen methanogen sequences (Methanobrevibacter spp.) to the peatland soil that seemed related to increased CH₄ production potential. Our findings indicate that cattle grazing of drained peatlands changes their methanogenic microbial community, may introduce rumen-associated methanogens and leads to increased CH₄ production. Consequently, rewetting of previously cattle-grazed peatlands has the potential to lead to increased CH₄ emissions. Careful consideration of land use history is crucial for successful climate mitigation with peatland rewetting.     

Bottom-up down from the top: Honeydew as a carbon source for soil organisms

Honeydew produced by aphids is a well-studied food source for mutualistic ants. Although considerable amounts drop onto the soil surface there are hardly any studies dealing with the potential effects of this additional C source for soil organisms, particularly those living at the soil surface. We studied this question by simulating honeydew and investigating plots under Populus canescens formerly infested with aphids on two ruderal sites differing in soil organic matter content and vegetation cover. The simulated honeydew input increased microbial biomass with some delay in week 10. Both natural and simulated honeydew raised the activity densities of an epigeic Collembola taxon, the Bourletiellidae, but not of the dominant Hemisotoma thermophila. The honeydew effect varied over time and with the site, and was more pronounced at the nutrient-poor site for Collembola, but not for microbial biomass. Ant consumption reduced the amount of honeydew reaching the soil surface by 50%. The activity density of H. thermophila was negatively related to ants and spiders, hinting at a top-down control. Honeydew clearly acts as a bottom-up force for soil organisms, which is however only part of the complex network connecting the food webs of primary producers and decomposers.     

Fast and efficient discrimination of the Isotoma viridis group (Insecta: Collembola) by PCR-RFLP

Identification of collembolan species is generally based on specific morphological characters, such as chaetotaxy and pigmentation pattern. However, some specimens do not match to described characters because these refer to adult specimens, often of one specific sex, or the characters are highly variable in adults (e.g. pigmentation, setae or furcal teeth). Isozymes have frequently assisted species discrimination, and also these may vary with developmental stage or environmental conditions. For identification of single species of the Isotoma viridis group, we present both direct sequencing of the cytochrome oxidase subunit II (COII) gene and a simple DNA-based molecular method.

Five PCR primers amplifying the COII region (717 bp) of the mitochondrial DNA were used. The sequences clearly separated the species I. viridis, I. riparia and I. anglicana, irrespective of colour varieties within the first species. DNA amplification products of different species can also be distinguished by digestion with restriction endonucleases, followed by gel electrophoresis for separation of fragments. This restriction fragment length polymorphism (RFLP), obtained after digestion with the endonucleases TaqI, VspI, MvaI and Bsp143I, revealed specific fragments that separated the three species from each other. Since restriction enzymes are sensitive to single base mutations, we suggest to use a combination of enzymes with at least two species-specific restriction sites when using the RFLP technique. For the I. viridis complex, VspI and Bsp143I appear to be an appropriate combination.     

Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

Sulfated polysaccharides (SP) from algae are of great interest due to their manifold biological activities. Obstacles to commercial (especially medical) application include considerable variability and complex chemical composition making the analysis and the quality control challenging. The aim of this study was to evaluate a simple microplate assay for screening the quality of SP. It is based on the fluorescence intensity (FI) increase of the sensor molecule Polymer-H by SP and was originally developed for direct quantification of SP. Exemplarily, 65 SP batches isolated from the red alga Delesseria sanguinea (D.s.-SP) and several other algae polysaccharides were investigated. Their FI increase in the Polymer-H assay was compared with other analytical parameters. By testing just one concentration of a D.s.-SP sample, quality deviations from the reference D.s.-SP and thus both batch-to-batch variability and stability can be detected. Further, structurally distinct SP showed to differ in their concentration-dependent FI profiles. By using corresponding reference compounds, the Polymer-H assay is therefore applicable as identification assay with high negative predictability. In conclusion, the Polymer-H assay showed to represent not only a simple method for quantification, but also for characterization identification and differentiation of SP of marine origin.     

Comparison of computer assisted surgery with conventional technique for treatment of abaxial distal phalanx fractures in horses: an in vitro study

Objective— To (1) evaluate and compare computer-assisted surgery (CAS) with conventional screw insertion (conventional osteosynthesis [COS]) for treatment of equine abaxial distal phalanx fractures; (2) compare planned screw position with actual postoperative position; and (3) determine preferred screw insertion direction.
Study design— Experimental study.
Sample population— Cadaveric equine limbs (n=32).
Methods— In 8 specimens each, a 4.5 mm cortex bone screw was inserted in lag fashion in dorsopalmar (plantar) direction using CAS or COS. In 2 other groups of 8, the screws were inserted in opposite direction. Precision of CAS was determined by comparison of planned and actual screw position. Preferred screw direction was also assessed for CAS and COS.
Results— In 4 of 6 direct comparisons, screw positioning was significantly better with CAS. Results of precision analysis for screw position were similar to studies published in human medicine. None of evaluated criteria identified a preferred direction for screw insertion.
Conclusion— For abaxial fractures of the distal phalanx, superior precision in screw position is achieved with CAS technique compared with COS technique.
Clinical Relevance— Abaxial fractures of the distal phalanx lend themselves to computer-assisted implantation of 1 screw in a dorsopalmar (plantar) direction. Because of the complex anatomic relationships, and our results, we discourage use of COS technique for repair of this fracture type.