Blue EaLAMP—a specific and sensitive method for visual detection of genomic Erwinia amylovora DNA

Fire blight caused by the gram-negative bacterium Erwinia amylovora is a major disease of pome fruit like apple (Malus domestica) or pear (Pyrus communis). Detection of the pathogen is hampered by low titres usually being present during initial flower infection and the brief time frame for analysis to decide upon subsequent countermeasures. Here we describe a loop-mediated isothermal amplification of DNA (LAMP) assay for genomic DNA of Erwinia amylovora, which relies on a highly specific primer design that excludes amplification from typical DNA sources of the orchard biological system and from sample handling. The assay enables the fast detection of down to approximately 20?cfu of pure Erwinia amylovora or 100?fg genomic DNA (corresponding to approximately 25 E. amyolovora equivalents) per reaction within 45?min. Fast and reliable detection of E. amylovora in orchard samples by naked eye is achieved through a visual colour change indication with hydroxynaphthol blue. The assay avoids use of complex technical devices and is thus suitable for field testing.     

硫酸铜,硝酸银和脱落酸对草地早熟禾胚性愈伤诱导的影响

试验以草地早熟禾4个品种的成熟种子为外植体,研究了CuSO4、AgNO3、脱落酸(ABA)对草地早熟禾胚性愈伤诱导的影响,结果表明,K3诱导培养基中添加CuSO4对草地早熟禾胚性愈伤组织的形成有促进作用,除DP192-10之外的3个品种都发生了明显的形态上的变化,DP-LF-300和2W42-117在添加5μmol/LCuSO4的培养基上诱导出了乳白色、致密的胚性愈伤组织,2W42-117的胚性愈伤率高达40%。在MS基本培养基中添加AgNO3、ABA对草地早熟禾4个品种的胚性愈伤组织的诱导没有作用。     

Influence of ketamine or xylazine supplementation on isoflurane anaesthetized horses‐ a controlled clinical trial

ObjectivesTo determine the influence of ketamine or xylazine constant rate infusions on isoflurane requirements, cardiovascular parameters and quality of anaesthesia in horses undergoing elective surgery.Study designProspective, matched paired clinical trial.AnimalsFifty four adult Warmblood horses.MethodsAfter premedication with acepromazine, xylazine and butorphanol, anaesthesia was induced with ketamine-midazolam and maintained with isoflurane alone (I), isoflurane with either 1 mg kg⁻¹ hour⁻¹ ketamine (IK) or same dose of xylazine (IX). End tidal concentration of isoflurane (Fe’Iso) was adjusted by the same anaesthetist in all horses according to a scoring system. Dobutamine was infused to maintain mean arterial pressure (MAP) =70 mmHg. Arterial blood gases, heart rate (HR), respiratory rate, MAP and cardiac output (lithium dilution) were measured. Groups I and IK received xylazine before recovery. Recovery quality was scored.ResultsMean ± SD averaged Fe’Iso (volume%) was significantly lower in IX (0.95 ± 0.07) and IK (0.97 ± 0.08) than in I (1.16 ± 0.13). In group IX, HR was significantly lower and averaged MAP (90 ± 13 mmHg) significantly higher than in groups I (71 ± 7 mmHg) and IK (76 ± 7 mm Hg). Differences in other cardiopulmonary variables did not reach statistical significance. All horses recovered well with best score in group IX.ConclusionsBoth CRIs of xylazine and of ketamine resulted in pronounced reduction of isoflurane requirements and blood pressure support based on routinely monitored parameters. Cardiac output appeared well maintained in all three protocols, but lithium dilution induced errors mean the results are untrustworthy. The work requires repetition with another mode of measurement of cardiac output.Clinical relevanceAll three protocols provided good clinical anaesthesia with clinically acceptable cardiovascular effects.     

Studies on the effect of deltamethrin sprays on the numbers of epigeal predatory arthropods occurring in arable fields

The effect of spraying winter rape and winter wheat with deltamethrin on the numbers of epigeal Carabidae, Staphylinidae and Linyphiidae caught in pitfall traps was studied in Northern Germany in 1980 and 1981. Two experiments were performed with each crop, deltamethrin being sprayed at 7.5 g a.i. ha^?1 in 300 litres of water in springtime on rape, and in midsummer on wheat. Sprayed plots were rectangles of 2 and 3 ha; unsprayed plots were squares of 0.5 ha each (two replicates). Ten pitfall traps were placed per replicate at least 3 weeks before spraying and were left for at least 3 weeks after spraying, the traps being emptied weekly. Beneficial Carabid species which have been shown already to have an economic significance, were rare during this study, but four other species (three of them were autumn breeders) were not affected by deltamethrin. At the spraying dates the autumn breeders were hidden in the soil as pupae. Three Staphylinid species, studied in only one experiment each, appeared to be strongly reduced in numbers by deltamethrin. The Linyphiid spiders were affected most heavily by deltamethrin sprayed in spring or in midsummer; the initial mortality of more than 92% lasted longer than 4 weeks, and an adverse effect was still significant in winter wheat 6 weeks after spraying. Overall, deltamethrin appears to have a broad spectrum effect concerning the epigeal predatory arthropods.     

Effects of the aglycone of ascaulitoxin on amino acid metabolism in Lemna paucicostata

Ascaulitoxin and its aglycone (2,4,7-triamino-5-hydroxyoctanoic acid, CAS 212268-55-8) are potent phytotoxins produced by Ascochyta caulina, a plant pathogen being developed for biocontrol of weeds. The mode of action of this non-protein amino acid was studied on Lemna paucicostata. Ascaulitoxin is a potent growth inhibitor, with an I₅₀ for growth of less than 1 μM, almost completely inhibiting growth at about 3 μM. Its action is slow, starting with growth inhibition, followed by darker green fronds, and then chlorosis and death. Most amino acids, including non-toxic non-protein amino acids, reversed the effect of the toxin when supplemented in the same medium. Supplemental sucrose slightly increased the activity. d-Amino acids were equally good inhibitors of ascaulitoxin activity, indicating the amino acid effects may not be due to inhibition of amino acid synthesis. Oxaloacetate, the immediate precursor of aspartate, also reversed the activity. LC-MS did not detect interaction of the compound with lysine, an amino acid that strongly reversed the effect of the phytotoxin. Metabolite profiling revealed that the toxin caused distinct changes in amino acids. Reduction in alanine, paralleled by enhanced levels of the branched chain amino acids valine, leucine and isoleucine and nearly unchanged levels of pyruvate, might indicate that the conversion of pyruvate to alanine is affected by ascaulitoxin aglycone. In addition, reduced levels of glutamate/glutamine and aspartate/asparagine might suggest that synthesis and interconversion reactions of these amino group donors are affected. However, neither alanine aminotransferase nor alanine: glyoxylate aminotransferase were inhibited by the toxin in vitro. Our observations might be explained by three hypotheses: (1) the toxin inhibits one or more aminotransferases not examined, (2) ascaulitoxin aglycone affects amino acid transporters, (3) ascaulitoxin aglycone is a protoxin that is converted in vivo to an aminotransferase inhibitor.     

Dissolved and colloidal phosphorus fluxes in forest ecosystems—an almost blind spot in ecosystem research

Understanding and quantification of phosphorus (P) fluxes are key requirements for predictions of future forest ecosystems changes as well as for transferring lessons learned from natural ecosystems to croplands and plantations. This review summarizes and evaluates the recent knowledge on mechanisms, magnitude, and relevance by which dissolved and colloidal inorganic and organic P forms can be translocated within or exported from forest ecosystems. Attention is paid to hydrological pathways of P losses at the soil profile and landscape scales, and the subsequent influence of P on aquatic ecosystems. New (unpublished) data from the German Priority Program 1685 “Ecosystem Nutrition: Forest Strategies for limited Phosphorus Resources” were added to provide up‐to‐date flux‐based information. Nitrogen (N) additions increase the release of water‐transportable P forms. Most P found in percolates and pore waters belongs to the so‐called dissolved organic P (DOP) fractions, rich in orthophosphate‐monoesters and also containing some orthophosphate‐diesters. Total solution P concentrations range from ca. 1 to 400 µg P L^?1, with large variations among forest stands. Recent sophisticated analyses revealed that large portions of the DOP in forest stream water can comprise natural nanoparticles and fine colloids which under extreme conditions may account for 40–100% of the P losses. Their translocation within preferential flow passes may be rapid, mediated by storm events. The potential total P loss through leaching into subsoils and with streams was found to be less than 50 mg P m^?2 a^?1, suggesting effects on ecosystems at centennial to millennium scale. All current data are based on selected snapshots only. Quantitative measurements of P fluxes in temperate forest systems are nearly absent in the literature, probably due to main research focus on the C and N cycles. Therefore, we lack complete ecosystem‐based assessments of dissolved and colloidal P fluxes within and from temperate forest systems.     

Biological control of fusarium seedling blight disease of wheat and barley

ABSTRACT Fusarium fungi, including F. culmorum, cause seedling blight, foot rot, and head blight diseases of cereals, resulting in yield loss. In a screen for potential disease control organisms and agents, Pseudomonas fluorescens strains MKB 100 and MKB 249, P. frederiksbergensis strain 202, Pseudomonas sp. strain MKB 158, and chitosan all significantly reduced the extent of both wheat coleoptile growth retardation and wheat and barley seedling blight caused by F. culmorum (by 53 to 91%). Trichodiene synthase is a Fusarium enzyme necessary for trichothecene mycotoxin biosynthesis; expression of the gene encoding this enzyme in wheat was 33% lower in stem base tissue coinoculated with Pseudomonas sp. strain MKB 158 and F. culmorum than in wheat treated with bacterial culture medium and F. culmorum. When wheat and barley were grown in soil amended with either chitosan, P. fluorescens strain MKB 249, Pseudomonas sp. strain MKB 158, or culture filtrates of these bacteria, the level of disease symptoms on F. culmorum-inoculated stem base tissue (at 12 days post- F. culmorum inoculation) was >/=31% less than the level on F. culmorum-inoculated plants grown in culture medium-amended soil. It seems likely that at least part of the biocontrol activity of these bacteria and chitosan may be due to the induction of systemic disease resistance in host plants. Also, in coinoculation studies, Pseudomonas sp. strain MKB 158 induced the expression of a wheat class III plant peroxidase gene (a pathogenesis-related gene).     

Effect of diet on results obtained by use of two commercial test kits for detection of occult blood in feces of dogs.

To evaluate the effect of diet on results obtained by use of 2 commercial test kits for detection of occult blood in feces, 5 dogs were fed 7 diets in randomized sequence. Dry and canned diets with various principal ingredients were evaluated. Each diet was offered twice over a 24-hour period, followed by a 36-hour nonfeeding period. Fecal specimens were collected twice daily, and tests for occult blood were performed within 12 hours. The dietary origin of fecal specimens was confirmed by use of colored markers fed with each diet, and was correlated with estimates of gastrointestinal tract transit time. A modified guaiac paper test and an o-tolidine tablet test were performed on each specimen. Of 59 specimens, 4 were positive for occult blood, using the o-tolidine tablet test. Three positive results were associated with a mutton-based canned diet, and 1 positive result was associated with a canned beef-based diet. Of 59 specimens, 11 were positive for occult blood, using the modified guaiac paper test. Four positive results were associated with the mutton diet, and 3 positive results were associated with the beef diet. Of the remaining 5 diets, 4 caused 1 positive reaction. Results were inconsistent with the null hypothesis that the distribution of positive occult blood test results is not affected by diet (P < 0.025), and indicate that diet can affect the specificity of peroxidase-based tests for detection of occult blood in canine feces. Diet modification prior to testing is recommended.     

德国及蒙古沙棘品种无性系生物学和观赏性价值评估

沙棘由于其有丰富的内含物和多方面的用途正被作为＂新兴水果＂而赢得了人们越来越多的重视和喜爱.通过在柏林Baumschulenweg的育种工作,到目前为止,已经育成不同成熟期的5个雌性和4个雄性沙棘品种.更多的由欧洲和蒙古原生种产生的杂交选育工作仍在进行中.将这些无性系按照其生物学和果实性状进行评估.就单个性状来讲,各品种之间的差异非常大（比如维生素的含量在17～340mg/100g之间变动,胡萝卜素的变动范围为6.5～23.8mg/100g）.这表明,进一步的育种改良完全有可能.所有用于比较的沙棘品种由在德累斯顿Pillnitz进行的育种工作获得,这些品种将被进一步研究.为了保险起见,以及为了解决后来有可能产生的育种及植物病理学方面的问题,必须尽可能保存现有的所有的某一品种的基因多样性.为此目的,由国家资助扶持的基因库的建立是很有必要的.