Results from a German interlaboratory test to establish the broth microdilution method for the determination of the minimum inhibitory concentration of bacteria from animals

In accordance with NCCLS guideline M31-A2, the DVG working group "antimicrobial resistance" developed a standard operating procedure (SOP) for the determination of the minimal inhibitory concentration (MIC) of antimicrobial agents by broth microdilution. This SOP was evaluated for its fitness for use in a national interlaboratory test. A total of 32 participating laboratories tested five strains (including two internationally accepted reference strains and three field strains representing in total three different bacterial species) three times at a one week interval each, using uniform microtitre plates. In 31 of the 32 laboratories more than 80% of MIC determinations performed yielded values in the expected range. In total 94.0% of the results were reproducible, with a lesser deviation of 4.0% from the expected values for laboratories performing MIC determination as a matter of routine (46.9%), compared to 7.9% for laboratories without such routine (53.1%). Comparing the consistency of results on the basis of the tested strains, a higher reproducibility of the results was observed for reference strains (96.1%) than for field strains (92.6%). In particular results obtained for the Streptococcus uberis field strain were afflicted with a higher error ratio (98 deviations from the expected values). Among the tested antimicrobial agents, a higher variability of results was recorded only for gentamicin with 16.7% divergent MIC determinations (mean value 6.0%). The high reproducibility of the results confirmed by this interlaboratory study underlines the robustness of the developed SOP as well as broth microdilutions as the method of choice for MIC determina tion.     

Cross-resistance between antimicrobial agents used in veterinary medicine: molecular background and practical consequences for susceptibility testing

Phenotypic resistance of veterinary pathogens to more than one antimicrobial agent (multi-resistance) may be caused by intrinsic resistance to the antimicrobial agents, acquired cross-resistance, or acquired co-resistance. Known cross-resistances allow to select so-called "representative substances" which are tested and the results of which can also be regarded as being valid for other members of the same class of antimicrobial agents. In general, a limitation in the number of antimicrobial agents to be tested in routine diagnostics is necessary because of capacity and cost efficiency. This is of particular relevance when the broth microdilution method - recommended as the method of choice - with 96-well microtiter plates is used. The knowledge about the relationship between different resistance phenotypes and the corresponding resistance mechanisms is of major value for both, the laboratory personnel and the veterinary practitioner. This review explains how "representative substances" for the most relevant classes of antimicrobial agents used in veterinary medicine are chosen on the basis of known cross-resistances.     

连续5年施用氮肥和羊粪的内蒙古羊草（Leymus chinensis） 草原土壤颗粒状有机质特征

 【目的】颗粒状土壤有机质（particulate organic matter,POM）是土壤有机质比较活跃的成分,受施肥、耕作及管理等多种因素的影响,本文研究连续5年施用氮肥和羊粪对内蒙古草原围栏封育的表层土壤颗粒状有机质的含量及其化学结构的影响。【方法】采用物理分组法获得游离态颗粒状有机质（FPOM）和包被态颗粒状有机质（OPOM）,并用13C核磁共振法测定其化学结构。【结果】施用氮肥的土壤pH值降低了0.25～0.78个单位,微生物量碳降低了13%～27%,POM含量没有明显变化;而施用羊粪的土壤pH提高了0.47～0.59个单位,微生物量碳提高了19%,POM大幅度提高,特别是FPOM最多提高了96%。OPOM的化学结构几乎不受施肥的影响,但FPOM的化学结构在一定程度上受到施肥特别是羊粪的影响,烷基碳增多,而羰基碳减少。【结论】长期施肥特别是有机肥料,不仅引起草原土壤颗粒状有机质含量的变化,还导致其化学结构的改变,FPOM组分变化更加明显。     

Quantification of Wheat spindle streak mosaic virus and Soil borne cereal mosaic virus in resistance testing of field samples of triticale using real-time RT-PCR

Journal of Plant Diseases and Protection - Triticale, a cross between rye and wheat, is a crop important for animal feed and the production of biogas and ethanol. Soil-borne viruses found in wheat...     

Glucosinolate concentration in turnip (Brassica rapa ssp. rapifera L.) roots as affected by nitrogen and sulfur supply

Three greenhouse pot experiments were conducted with four different nitrogen (N) treatments (80, 160, 240, and 320 kg ha (-1)) in combination with three sulfur (S) treatments (10, 20, and 60 kg ha (-1)) to study the effects of combined N and S supply on glucosinolate concentration and composition in turnip roots. Total glucosinolate concentration varied widely from 9.7 (N 320S 10) to 91.6 (N 160S 60) mg (100 g) (-1) root fresh weight (FW) and individual glucosinolate concentrations were increased with increasing S supply regardless of the N treatment, whereas enhanced N supply (160 - 320 N ha (-1)) at the high S level (60 kg ha (-1)) did not affect total glucosinolate concentration. In contrast, assumingly attributed to the individual glucosinolate biosynthesis concentration of N-containing tryptophan-derived indole glucosinolate was highest with increased N supply, whereas S-containing methionine-derived aromatic and aliphatic glucosinolates decreased with increasing N supply combined at low S level (10-20 kg ha (-1)).     

Influence of nitrogen forms and mycorrhizal colonization on growth and composition of Chinese bunching onion

In recent years, interest has grown in cultivating Allium species with enhanced health benefits and/or distinct flavor. Concentrations of phytochemicals determining these desired characteristics may be influenced by nitrogen forms (ammonium or nitrate) and arbuscular mycorrhizal (AM) fungi. We examined these relations with the test plant bunching onion (Allium fistulosum L.). Three different ammonium‐to‐nitrate (NH : NO ) ratios were supplied in combination with or without inoculation with an AM fungus (Glomus mosseae). The plants were evaluated for dry weight, leaf number, and content of nutrients (N, NO , P, S), sugars (glucose, fructose, and sucrose), and organosulfur compounds (measured as pyruvic acid). The experiment was carried out under controlled conditions in a greenhouse. Plants were grown on perlite amended twice a day with nutrient solution. In nonmycorrhizal plants, the application of nutrient solution with predominant NO or NH₄NO₃ as N source supported adequate growth of Allium fistulosum while predominant NH supply resulted in decreased growth and occurrence of wilting symptoms. Mycorrhizal inoculation significantly increased dry weight and leaf number of predominantly NH ‐fed or NH₄NO₃‐fed plants. While shoot P concentration increased with higher NH supply, shoot N concentration increased in predominantly NH ‐fed plants only. Nitrogen form and AM colonization had little effect on shoot S or sugar concentrations. The total content in organosulfur compounds was significantly affected by both, N form and AM colonization. The optimal growth condition for a high formation of organosulfur compounds in this experiment was a nutrient solution with predominant NO supply, but when supported by AM fungi, Allium fistulosum produced similar amounts of pyruvic acid in NH₄NO₃‐fed plants.     

Idiopathic acute onset myelopathy in cheetah (Acinonyx jubatus) cubs.

Numerous cases of ataxia, hind limb paresis, and paralysis have occurred in cheetah (Acinonyx jubatus) cubs over the past 10 yr within the European Endangered Species Program population, including 12 in mainland Europe, two in the British Isles, one in Namibia, and one in Dubai. The condition is the most important medical factor limiting European cheetah population growth. Eight cubs at the Salzburg Zoo, Austria, were affected. They demonstrated upper motor neuron lesions when alive and bilateral, symmetrical myelin degeneration of the spinal cord on necropsy. Ballooning of myelin sheaths surrounded mostly preserved axons, and no spheroids, characteristic of acute axonal degeneration, were found. Myelin loss markedly exceeded axonal degeneration. The syndrome's etiology is unclear, although viral, bacterial, parasitic, genetic, nutritional-metabolic, toxic, and physical causes have been considered.     

Microbial community development in the rhizosphere of apple trees at a replant disease site

Apple replant disease (ARD) is a complex syndrome that affects young trees in replanted orchard sites causing necrotic lesions on feeder roots, stunted tree growth and reduced cumulative yields. Use of ARD-tolerant rootstocks is an emerging control strategy. We studied the bacterial, fungal, and oomycetes populations in the rhizosphere of five rootstock cultivars (M.7, M.26, G.16, G.30 and CG.6210) planted into the old tree row or grass lanes of a previous orchard in Ithaca, NY, to better understand the role of rhizosphere microbial communities in the prevalence and control of ARD. The possible involvement of antagonistic Pseudomonas species, Pythium spp., Phytophthora spp. and rhizosphere cyanide concentrations in ARD were also examined. The rootstocks M.7, M.26 and G.16 were susceptible to ARD, while G.30 and CG.6210 were more tolerant. Tree growth on the rootstocks M.7, M.26 and G.16 was reduced by 10% when planted in the old tree rows, but this did not significantly reduce yields in the first fruiting year. The susceptible rootstocks, M.7 and M.26, supported higher densities of culturable rhizosphere fungi and bacteria than G.16, G.30 and CG.6210. Over 2 years, microbial densities were highest in July, lower in May and lowest in September. The composition of bacterial and fungal communities in the rhizosphere was highly variable and changed over seasons and years, as assessed by terminal restriction fragment length polymorphism (T-RFLP) analyses. Initial differences in fungal rhizosphere communities between the two planting positions converged 2 years after the trees were replanted. In contrast, the bacterial rhizosphere community composition still differed significantly between the two planting positions 3 years after the orchard was replanted. The bacterial and fungal rhizosphere community compositions of susceptible rootstocks, M.7 and M.26, differed from those of the tolerant rootstocks, G.30 and CG.6210; G.16, differed from all the other rootstocks. The observed effects of rootstocks, planting positions and time on microbial community composition were small relative to the high variability observed overall. Pythium spp. and Phytophthora spp. infestations were high and similar for all rootstocks and planting positions. Neither potentially antagonistic Pseudomonas nor rhizosphere cyanide concentrations appeared to be involved in the ARD-complex at the studied site. Avoiding replanting into the old tree rows coupled with use of tolerant rootstocks appear to be the best strategies for reducing ARD in replanted orchards. Changes in rhizosphere microbial communities are among the many factors that contribute to improved tree growth when these management strategies are used.