Replication characteristics of swine influenza viruses in precision-cut lung slices reflect the virulence properties of the viruses

Precision-cut lung slices of pigs were infected with five swine influenza A viruses of different subtypes (A/sw/Potsdam/15/1981 H1N1, A/sw/Bad Griesbach/IDT5604/2006 H1N1, A/sw/Bakum/1832/2000 H1N2, A/sw/Damme/IDT5673/2006 H3N2, A/sw/Herford/IDT5932/2007 H3N2). The viruses were able to infect ciliated and mucus-producing cells. The infection of well-differentiated respiratory epithelial cells by swine influenza A viruses was analyzed with respect to the kinetics of virus release into the supernatant. The highest titres were determined for H3N2/2006 and H3N2/2007 viruses. H1N1/1981 and H1N2/2000 viruses replicated somewhat slower than the H3N2 viruses whereas a H1N1 strain from 2006 multiplied at significantly lower titres than the other strains. Regarding their ability to induce a ciliostatic effect, the two H3N2 strains were found to be most virulent. H1N1/1981 and H1N2/2000 were somewhat less virulent with respect to their effect on ciliary activity. The lowest ciliostatic effect was observed with H1N1/2006. In order to investigate whether this finding is associated with a corresponding virulence in the host, pigs were infected experimentally with H3N2/2006, H1N2/2000, H1N1/1981 and H1N1/2006 viruses. The H1N1/2006 virus was significantly less virulent than the other viruses in pigs which was in agreement with the results obtained by the in vitro-studies. These findings offer the possibility to develop an ex vivo-system that is able to assess virulence of swine influenza A viruses.     

Extraction and characterization of soil hydrogenases oxidizing atmospheric hydrogen

Soils are the largest sink of atmospheric hydrogen contributing about 75% to the total budget. Atmospheric H₂ is assumed to be oxidized in soil by abiontic soil hydrogenases. Extraction of a forest soil with a slightly alkaline (pH 8.5) buffer containing polyethylene glycol (PEG), followed by filtration yielded a bacteria-free extract that oxidized H₂ at ambient concentrations (0.2–2.0 ppmv). Hydrogenase activity was assayed by gas chromatographic analysis of H₂ consumption and by conversion of ³H₂ to tritiated water. Only less than 2% of the original activity was recovered in the extract. Kinetic analysis nevertheless resulted in a biphasic kinetics exhibiting two K_m and V_max values that were similar to those detected in the original soil. In addition, activities of both original soil and soil extract showed similar optima at pH 4–6 and at 30 °C, indicating that representative fractions of soil hydrogenases were recovered in the extract. Precipitation with PEG or ultrafiltration allowed further purification of the activity, albeit only about 20% of that in the crude extract could be recovered in the precipitate or the fraction >100 kDa.     

Comparison of arterial and central venous cannulations using ultrasound guidance in pigs

OBJECTIVE: To evaluate the feasibility of ultrasound guided vascular access in pigs by comparing central venous and arterial cannulation techniques. ANIMALS: Twenty-two healthy female Pietrain pigs, 14-18 weeks old and weighing 51.1 +/- 4.3 kg (mean +/- SD). STUDY DESIGN: Comparative animal trial. MATERIALS AND METHODS: After induction of general anaesthesia, cannulation of the external jugular vein and internal carotid artery was attempted using real-time ultrasound guidance. The quality of the ultrasound picture was assessed on an analogue scale from 1 (excellent) to 5 (insufficient). Vessel size, cannulation success rate, number of puncture attempts and time from first puncture attempt until insertion of the Seldinger wire were recorded. RESULTS: Cannulation was successful in all but one animal in which a cut-down technique was performed. The arteries were significantly smaller than the veins (p < 0.001) resulting in a significantly prolonged cannulation time (p = 0.032) for insertion of arterial catheters without differences in success rate. In 89% of attempted cannulations, the Seldinger wire was inserted within 5 minutes. CONCLUSIONS AND CLINICAL RELEVANCE: In anaesthetized pigs undergoing instrumentation for biomedical research, ultrasound-guided vascular access is a simple and rapid alternative to surgical cut-down. In veterinary anaesthesia, the technique might be useful in sedated or anesthetized pigs in which arterial or central venous access is required.     

Genetic diversity and population structure of burbot Lota lota in Germany: Implications for conservation and management

The genetic structure of the gadiform fish species, burbot Lota lota L., was investigated across Germany to derive management options for facilitating the preservation of genetic diversity. Sequence analysis of the mitochondrial control region (n = 244) and microsatellite analysis (n = 861) of specimens from 20 sites revealed genetic structuring between major river basins, and particularly between lake and river habitats. The admixture zone between the Eurasian and West European phylogenetic clades in Lake Constance was confirmed and expanded to include the drainage basins of the rivers Rhine and Schlei/Trave. Haplotype distribution and private haplotypes in single river basins indicated population differentiation and imply that German burbot constituted an important part of the entire species' diversity. The derived genetic structuring has implications for future stocking programmes and the preservation of the adaptive potential of burbot, a guiding species for oligotrophic lakes in Europe.     

间隙灌溉和控释肥施用对稻田土壤产甲烷微生物的影响

间隙灌溉和控释肥施用影响稻田CH_4的产生和排放,然而其微生物机理尚不清楚。本研究通过采集稻季田间原位试验新鲜土样,采用核酸定量技术(qPCR)和末端限制性片段长度多态性(T-RFLP)技术,研究间隙灌溉和控释肥施用对稻田土壤产甲烷微生物群落丰度和结构的影响。结果表明,稻季CH_4排放量与古菌、产甲烷菌(mcr A基因)和甲烷氧化菌(pmo A基因)数量均呈极显著正相关关系(P0.01),而与细菌数量无显著相关性。间隙灌溉显著影响产甲烷菌和甲烷氧化菌数量的季节变化,其中烤田抑制产甲烷菌生长,而对甲烷氧化菌数量没有显著影响。与尿素相比,施用控释肥增加了稻田土壤细菌、古菌和产甲烷菌数量,降低了甲烷氧化菌数量。土壤古菌群落的优势T-RFs片段为184bp和391bp,其中184bp片段的相对丰度随着间隙灌溉的进行由45%~55%降低到23%~30%;而391bp片段则相反,其相对丰度由12%~18%增加到23%~26%。典型相关性分析(CCA)表明间隙灌溉显著影响土壤古菌群落结构(P0.001),而控释肥施用对土壤古菌群落结构没有明显影响。     

Ligands of the nicotinic acetylcholine receptor as insecticides

Insect nicotinic acetyl receptors (nAChR) are targets of growing importance and, since the early 1990s, the number of such highly effective insecticides as imidacloprid and spinosyn has grown. Several natural compounds, eg dihydro-β-erythroidine, methyl caconitine and paraherquamide, showing high affinity to the same receptor, were considerably less active as insecticides, most likely because of their antagonistic action. Our observations on aphids after ingestion of the antagonistic compound dihydro-β-erythroidine revealed anti-feedant-like properties. As a consequence, the symptomology of poisoning was totally different between agonists and antagonists of the nAChR. Electrophysiological (whole-cell voltage clamp) measurements in isolated housefly neurones revealed that agonism seems to be a prerequisite for insecticidal activity. Furthermore, we were able to demonstrate the existence of two different subtypes of the nAChR in isolated locust neurones with different pharmacology and ion-channel properties.     

Cold tolerance during juvenile development in sorghum: a comparative analysis by genomewide association and linkage mapping

Improved cold tolerance during the juvenile phase is a major breeding goal to develop new sorghum cultivars suitable as an alternative energy crop in temperate regions. The objectives of this study were to identify marker‐trait associations for cold tolerance in a sorghum diversity panel fingerprinted with 2620 single nucleotide polymorphism (SNP) markers and to detect quantitative trait loci (QTL) in two F_2:3 populations. Traits of interest were dry matter growth rate (DMGR), leaf appearance rate (LAR), chlorophyll content (SPAD) and chlorophyll fluorescence (F_v′/F_m′ and Ф_PSII) in relation to temperature. The association panel comprised 194 genotypes, while the F_2:3 populations consisted of 80 and 92 genotypes. All populations were tested under controlled conditions in a minimum of four temperature regimes ranging from 9.4°C to 20.8°C. QTL were identified for means across environments and regression parameters describing temperature effects. Several marker‐trait associations for mean (m) DMGR, base temperature (T_b) of SPAD and Ф_PSII and temperature effect on LAR were validated by QTL detected in population 1 or 2. Promising QTL regions were found on chromosomes SBI‐01, SBI‐02, SBI‐03, SBI‐04, SBI‐06 and SBI‐09, among them genomic regions where candidate genes involved in the C‐repeat binding pathway or encoding cold‐shock proteins are located.